application notes nov2015a - obrnutafaza.hr · 37.5°c 40°c 42.5°c 1 1. ......

Chromatography Applications

HPLC | IC | SPE

discoverysciences.com

Applications

HPLC Applications . . . . . . . . . . . . . .

Pharmaceuticals 1–17

Nutraceuticals/Vitamins 18–24

Food & Beverage 25–38

Life Science 39–52

Environmental 53–57

Miscellaneous 58–63

IC Applications . . . . . . . . . . . . . . . .

Anions 64–66

Cations 67–68

SPE Applications . . . . . . . . . . . . . . . 69–79

Pharmaceuticals 69

Nutraceuticals 70–71

Food & Beverage

Life Science 74

Environmental 75–79

72–73

64–68

1–63

0 2 4 6 8

1

2

3 4

0 2 4 6 8 10 12 14 M0 2

1

2 3

4

5

6

4 6 8

0

2

1

2.5 5 7.5 10 12.5 15 17.5

Column: Alltech® Platinum™ C18, 5µm, 150 x 4.6mm

(Part No. 32043)

Mobile Phase: 0.02M KH2PO4:Methanol:Acetonitrile:

pH4.5 (40:50:10)

Flow Rate: 1.0mL/min

Detector: UV at 254nm

1. Naproxen 2. Acemetacin 3. Indomethacin 4. p-Xylene

Column: Alltech® Prevail™ Amide, 5µm, 150 x 4.6mm

(Part No. 88660)

Mobile Phase: 0.02M K2HPO4, pH7.0:Acetonitrile (75:25)



1. Aspirin 2. Acetaminophen 3. Naproxen 4. Fenoprofen

938510124

1. Aspirin 2. Acetaminophen 3. Naproxen 4. Fenoprofen 5. Ibuprofen 6. Diclofenac

Column: ® Alltima C8, 5µm, 150 x 4.6mm (Part No. 88072)

Mobile Phase: A: 0.05M K2HPO4, pH3.3 B: Methanol

C: Acetonitrile

Gradient: Time: 0 2 5

%B: 10 10 30

%C: 10 10 25



Column: HP C8, 5µm, 150 x 4.6mm

(Part No. 87751)

Mobile Phase: 0.05M Phosphate Buffer pH3.0:Methanol (80:20)



1. Procainamide 2. Acetaminophen 3. Theophylline 4. Caffeine

Column: GraceSmart™ C18, 5µm, 150 x 4.6mm

(Part No. 5138812)

Mobile Phase: Phosphoric Acid in Water, pH2.5:Acetonitrile (65:35)

Flow Rate: 2mL/min


Injection Vol: 5µL

1. Valerophenone 2. Ibuprofen

Analgesics

Anti-Inflammatories

AnalgesicsAnalgesics

Basic Drugs

Ibuprofen (U.S.P.)

8156 10702

10374

0 1

1

2 3 Min.

1

1

1

1

30°C

32.5°C

35°C

37.5°C

40°C

42.5°C1

1. Ibuprofen (100ppm)

Column: µm, 2.1 x 100mm

(Part No. 87669)

Mobile Phase: Acetonitrile: 0.1%TFA in Water (80:20)


Column Temp: Ambient

Semi-Volatiles

Detector: ELSD

hplc applicationspharm

aceuticals

1 www.discoverysciences.com

®

®

Alltech

® Alltima ®Alltech

HP C18, 3HP C18, 3® Alltima ®Alltech

10 Min. 20 Min.

10 Min. Min.

Min.

0 1 2 3 4 5 Min.

1

2

3

Column: Alltech® Prevail™ C18, 5µm, 150 x 4.6mm (Part No. 99208)

Mobile Phase: Water:Acetonitrile (10:90)


Detector: UV at 254

1. Methyl Paraben 2. Propyl Paraben 3. Hydrocortisone Acetate

9463

Hydrocortisone Acetate Cream

Column: GraceSmart™ C18, 5µm, 250 x 4.6mm (Part No. 5138810)

Mobile Phase: Phosphate Buffer:Methanol (80:20)



10770

Paracetamol Effervescent Tablets on Chinese Pharmacopeia Assay

Analgesics

mA

U

2000

1250

1000

1500

1750

750

500

250

0

5 6 7 83 41 2 9 10 Min.0

1. Paracetamol

0 1 2 3 4 5 6 7 8 9 10 Min.

2

1

Alltima HP

HILIC Columns

1

2

Reverse-Phase

C18 Columns

1. Albuterol

2. Bamethan

Basic Drugs

Column: Alltech® Prevail™ C18, 3µm, 53 x 7mm Rocket™

(Part No. 99279)

Mobile Phase: Water:Methanol (10:90)


Detector: UV at 254

1. Hydrocortisone 2. Testosterone 3. Progesterone

9392

Steroids

Steroids

10391

Column: HP CN, 5µm, 150 x 4.6mm

(Part No. 87769)

Mobile Phase: Hexane:Isopropanol (93.7:6.3)



1. Deoxycorticosterone Acetate

2. Hydrocortisone Acetate

3. Cortisone Acetate

4. Cortisone

5. Prednisone

6. Hydrocortisone

7. Prednisolone

Corticosteroids

Column: Prevail™ C18 Column, 5µm, 4.6 x 150mm

(Part No. 99208)

Alltima HP HILIC Column, 5µm, 4.6 x 150mm,

(Part No. 86464)

Mobile Phase: 0.05M Ammonium Formate, pH 3.0:CH3CN (88:12)

Acetonitrile:0.05M Ammonium Formate, pH 3.0 (85:15)



10949


aceutic

als

2www.discoverysciences.com

® Alltima ®

Alltech

®

®

1

2

0 20 Min.102 4 6 8 12 14 16 18

0 2

1

2

3

4

5

6

7

4 6 Min.

0 1 2 3 4 5 6 7 8 9 Min.

1

2

3

8957

Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™

(Part No. 50605)

Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water

B: 0.1% Trifluoroacetic Acid in Acetonitrile

Gradient: Time: 0 6

%B: 10 90


Detector: ELSD

1. Theophylline 2. Acetylsalicylic Acid 3. Propranolol 4. Nortriptyline 5. Trimipramine

Column: Silica, 5µm, 300 x 4.6mm (Custom)

Mobile Phase: Acetonitrile:Chloroform:i-Butyl Alcohol:Water:Glacial

Acetic Acid (85:8:5:2:0.05)



1. Triazolam 2. Alprazolam

8691

Column: Alltech® Platinum™ EPS C18, 3µm, 53 x 7mm Rocket™

(Part No. 50573)

Mobile Phase: 0.05M Ammonium Acetate, pH5.5:Acetonitrile (65:35)



9105

10373

Column: CN, 3µm, 150 x 4.6mm

(Part No. 81189)

Mobile Phase: 0.01M KH2PO4, pH2.6:Acetonitrile (50:50)



1. Desipramine 2. Nortryptyline 3. Diphenhydramine 4. Amitriptyline

7368

1. Diphenhydramine 2. Nortriptyline 3. Amitriptyline

Column: Alltech® Alltima HP C18 Amide, 5µm, 150 x 4.6mm

(Part No. 87733)

Mobile Phase: 0.025M Potassium Phosphate Buffer, pH7.0:Methanol

(15:85)

Flow Rate: 1mL/min


Column: Alltech® Alltima HP C8, 5µm, 150 x 4.6mm

(Part No. 87751)

Mobile Phase: A: 0.025M Potassium Phosphate Buffer, pH7.0

B: Methanol


%B: 60 75 75



1. Fluphenazine HCl 2. Chlorpromazine HCl 3. Phenothiazine HCl

1. Chlordiazepoxide Degradant 2. Nitrazepam 3. Norchlordiazepoxide 4. Nordiazepam

Universal High Speed Gradient

10353

Benzodiazepines and Metabolites

Tricyclic Antidepressants

Alprazolam and Triazolam by the USP XXII Method

Antidepressants

Basic Drugs

Tricyclic Amines

5. Chlordiazepoxide 6. Chlordiazepoxide Degradant 7. Diazepam


aceuticals


® Alltima ®Alltech ® Alltima ®Alltech

Alltima® ®

®

Min. Min.

Min. Min.

Min. Min.

3

12

3

4

0 1 2 3 4 5 6 7 8 Min.

1

2

3

4

0 1 2 3 4 5 6 7 8 Min.

2

3

1

45

6 7

0 2 4 6 8 10 12 14 16 18 Min.

1072010719

1. Uracil 2. Propanolol 3. Naphthalene 4. Dipropylphthalate

Column: Grace® VisionHT C18, 1.5µm, 50 x 2.0mm

(Part No. 5139605)

Mobile Phase: 20mM Potassium Phosphate, pH2.5:Methanol (40:60)



Temperature: Ambient

PropanololPropanolol

1. Uracil 2. Propanolol 3. Naphthalene 4. Dipropylphthalate

Column: Grace® VisionHT C18-P, 1.5µm, 50 x 2.0mm

(Part No. 5139606)




Temperature: Ambient

Column: Jones Genesis® C18, 4µm, 150 x 4.6mm

(Part No. FM15960E)

Mobile Phase: Acetonitrile:50mM KH2PO4, pH2.4 (35:65)



Basic Drugs

1. Amiloride 2. Propranolol 3. Chloramphenicol 4. Carbamazepine 5. Trimipramine 6. Trifluoperazine

10604

Column: Alltech® Alltima HiLoad, 5µm, 250 x 4.6mm

(Part No. 87698)



Detector: ELSD

1. Phenobarbital 2. Butabarbital 3. Butalbital 4. Pentobarbital 5. Amobarbital 6. Mephobarbital 7. Secobarbital

Barbiturate Mix

Antidepressants

BarbituratesBarbiturates

Column: C18, , 50 x 2.1mm

Mobile Phase: 40% CH 3CN:50mM Ammonium Acetate, pH5.4

Flowrate: 0.5mL/min


Phenobarbital

Amobarbital

Mephobarbital

5

1. Barbital

2.

3. Talbital

4.

5.

0 1 2 Min.

1

2

3

4

0 1 2 3 Min.

1

23

1. Acetylsalicylic Acid 2. Diphenhydramine 3. Trimipramine

Column: Alltech® Alltima µm,

7 x 33mm Rocket™ (Part No. 87671)

Mobile Phase: A: 0.1% TFA in Water B: 0.1% TFA in Acetonitrile

Gradient: Time: 0 3

%B: 10 90


Column Temp: 30°C

Stable Baselines with Fast, Steep Gradients

10658

9433

3µmAlltech® Alltima


aceutic

als


® ®

HP C18, 3®

® ®

pharm

aceuticals

Barbiturates

Column: Platinum™ EPS 100Å C18, 5µm, 150 x 4.6mmMobile Phase: 0.010M Sodium Acetate, pH4.0 Acetonitrile

(75:25)Flowrate: 1.0mL/minDetector: UV at 230nm

Column: Platinum™ EPS 100Å C18, 3µm, 53 x 7mmMobile Phase: 0.010M Sodium Acetate, pH4.0 Acetonitrile

(75:25)Flowrate: 3.0mL/minDetector: UV at 230nm

1. Barbital2. Butalbital3. Aprobarbital4. Talbutal5. Butabarbital6. Phenobarbital7. Amobarbital8. Mephobarbital

Run TimeReducedby 70%

7

12

3

456

8

0 4

12

3

4 5 67

8

8 12 Min.

ock

o o

0 1

12

3

4 5 67

8

2 3 4 Min.

8684

Rocket™ Column

0 2 4 6 8 10 12 14 16 18 20 Min.

2

1

UV at 230nm

21

ELSD

1. Mephobarbital (500ppm) 2. Secobarbital (500ppm)

10616

Column: Alltech® Alltima µm, 4.6 x 250mm

(Part No. 87698)

Mobile Phase: Acetonitrile:Water (30:70)


Column Temp: 45°C

1:1 Mixture

10617

10375

Column: Alltech® Alltima HP C8, 5µm, 250 x 4.6mm

(Part No. 87751)

Mobile Phase: 0.05M Ammonium Acetate:Acetonitrile (78:22)



1. Barbital 2. Phenobarbital 3. Talbutal 4. Amobarbital 5. Mephobarbital

Barbiturate Mix

Standard Platinum™ C18

Platinum™ EPS C18

Vasodilators

Column: 5µm, 4.6 x 150mm

Mobile Phase: 0.025M KH2PO4, pH 3.0:Methanol (50:50)



1. Pindolol 2. Pentoxifylline 3. Isoxsuprine 4. Nifedipine Degradant 5. Nifedipine

8387

8388

Vasodilators

8685

Barbiturates

hplc applications


HP C18 HL, 5®®

4

pharm

aceutic

als

hplc applications

10348

10349

Column: ® Alltima HP EPS C18, 5µm, 150 x 4.6mm

(Part No. 87715)



Detector: ELSD

Column: Alltech® Alltima HP C18 HiLoad, 5µm, 150 x 4.6mm

(Part No. 87697)



Detector: ELSD

Carbamazepine

1. Ethotoin 2. Phenytoin

Mephenytoin Carbamazepine

Column: Alltech® Prevail™ Phenyl, 5µm, 150 x 4.6mm

(Part No. 99239)

Mobile Phase: A: Water B: Acetonitrile

Gradient: Time: 0 15

%B: 25 60


Detector: UV at 254

Anticonvulsants

Anticonvulsants

Anticonvulsants

Anticonvulsants

9394

1. Ethotoin 2. Mephenytoin 3. Carbamazepine 4. Diazepam

0 2 4 6 8 10 12 14 16 18 20 Min.

1

2

3

4

1. Ethotoin 2. Phenytoin 3. Mephenytoin 4.

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.

1

2

3 3.

4.

1. Ethotoin

2. Phenobarbital

3. Methsuximide

4. Mephenytoin

5. Carbamazepine

6. Diazepam

123

4

5

6

0 20 Min.4 8 1612

Column: Phenyl,

Mobile Phase:

Gradient 0 20 25 30

%B: 15 60 60 15



5,5-DiphenylhydantoinCarbamazepine

Column: Platinum™ EPS 100Å C18, 3µm,53 x 7mm Rocket™ Column

Mobile Phase: 0.025M Ammonium Acetate, pH 5.5: Acetonitrile (70:30)Flowrate: 3.0mL/minDetector: Alltech ELSD

1. Ethotoin2. Mephenytoin3.

4.

0 1

1

2

2

3

4

3 Min.

7233

8946

A: Water B: Acetonitrile: Time:

Anticonvulsants

Anticonvulsants


®

®® Alltima HP EPS C18, 5µm, 150 x 4.6mm®Alltech ® Alltima

®® Alltima ®Alltech 250 x 4.6mm5µm,

pharm

aceuticals

hplc applications

Column: Alltech® Alltima C8, 5µm, 250 x 4.6mm (Part No. 88076)

Mobile Phase: A: 0.05M Potassium Phosphate, pH3.2

B: Acetonitrile

Gradient: Time: 0 5 20 25

%B: 25 25 70 70

Flow Rate: 1.3mL/minDetector: UV at 220nm

1. Tripelennamine 2. Triprolidine 3. Cyclizine 4. Chlorcyclizine 5. Meclizine

Column: Alltech® Prevail™ Carbohydrate ES, 5µm, 250 x 4.6mm

(Part No. 35101)

Mobile Phase: Acetonitrile:0.1% Ammonium Carbonate, pH6.5 (80:20)


Detector: ELSD

102757232

Column: Vydac® Denali®, 5µm, 150 x 4.6mm (Part No. 238DE5415)

Mobile Phase: A: Acetonitrile:Water (20:80)

B: Acetonitrile:Water (95:5)

Gradient: Time: 0 8

%B: 40 90



Antihistamines

Antihistamines

OTC Antihistamine and Excipients

Estrogens

Estrogens

0 1 2 3 4 5 6 7 8 9 10 11 Min.

1

3

4 5

1. Diphenhydramine 2. PEG, Gelatin 3. Sorbitan 4. Glycerol 5. Sorbitol

1. Estriol 2. 17 -Ethylestradiol 3. Estrone 4. Diethylstilbestrol 5. Hexestrol 6. Product of Diethylstilbestrol Decomposition


®

0

12

3

4

5

2 4 6 8 10 12 Min.

1 + 2

3

0 0.5 1 1.5 2 2.5 3 Min.

1 2

3

0 0.5 1 1.5 2 2.5 3 Min.

pharm

aceutic

als

hplc applications

10118

1. p-Aminobenzoic Acid 2. Benzocaine 3. Tetracaine 4. Dibucaine 5. Butacaine


Mobile Phase: A: Water B: Methanol C: Acetonitrile

Gradient: Time: 0 7

%B: 15 45

%C: 15 35




(Part No. 5139605)




1. Procaine 2. Lidocaine 3. Tetracaine

10737 10738


(Part No. 5139606)




1. Procaine 2. Lidocaine 3. Tetracaine

Anesthetics

Anesthetic Mix

Basic Pharmaceuticals Basic Pharmaceuticals

1. Procaine 2. Benzocaine 3. Mepivacaine 4. Lidocaine 5. Tetracaine 6. Dibucaine


Mobile Phase: Methanol:50mM K2HPO4 in Water, pH7.0 (60:40)



Anesthetics

1. Hydromorphone 2. Oxycodone 3. Codeine 4. Ethylmorphine 5. Nalorphine


Methanol:20mM K2HPO4 in Water, pH7.0 (50:50)


Detector: UV at 254

Opiates

Opiates


® ®

®

0 1

1

2

2

3

3

4

5

6

4

5 Min.

0 2 4 6 8 10 12 14 16 18 20 22 24 Min.

1

2

3

4

0 2 4 6 Min.

1

2

3

5

4

pharm

aceuticals

hplc applications

Column: Alltech® Prevail™ C18, 3µm, 53 x 7mm (Part No. 99279)



Detector: UV at 220

1. Metoprolol 2. Oxprenolol 3. Propranolol

Column: ZirChrom® PBD, 3µm, 150 x 4.6mm (Part No. ZR03-1546)

Mobile Phase: 0.01M Triethylamine, 0.05M Tetramethylammonium

Hydroxide, pH12.2:Acetonitrile (65:35)


Detector: UV at 254

10354

10161

Column: Alltech® Alltima HP CN, 5µm, 150 x 4.6mm

(Part No. 87769)

Mobile Phase 0.02M KH2PO4, pH3.0:Acetonitrile (70:30)



9395

10356

1. Labetalol 2. Acebutolol 3. Oxprenolol 4. Lidocaine 5. Alprenolol 6. Propranolol

8160


(Part No. 87733)

Mobile Phase: 0.025M Potassium Phosphate, pH2.8:Methanol (90:10)



1. Codeine 2. Strychnine 3. Quinine 4. Quinidine

1. Diltiazem 2. Verapamil 3. Nicardipine 4. Nifedipine


(Part No. 32043)

Mobile Phase: 0.05M Ammonium Dihydrogen Orthophosphate, pH4.5:

Methanol:Acetonitrile (40:50:10)



1. Barbital 2. Phenobarbital 3. Butalbital 4. Oxazepam 5. Triazolam

SedativesSedatives/Hypnotics

Basic Drugs

Anti-Hypertensives

Antihypertensives

Calcium Channel Blockers Drug Mix

ELSD

UV

0

1

1 2 3 4 5 6 7 Min.

1. Ethinamate (5.0 mg/mL)

Column: Alltech® Alltima µm, 10 x 150mm

(Part No. 81102)



Column Temp: 25°C

Ethinamate

10438


C18, 5

® ®

®

pharm

aceutic

als

hplc applications

Column: Alltech® Platinum™ EPS C18, 100Å, 5µm, 150 x 4.6mm (Part No. 32214)Mobile Phase: Water:Acetonitrile:Methanol:Glacial Acetic Acid (60:10:30:1)Flow Rate: 1.0mL/minDetector: UV at 254nm

Methylxanthines

Amphetamines

Anti-Asthmatics

0 1

1

2

2

3

3 4

4 Min.

1001410427

Column: Alltech® Alltima Silica, 3µm, 100 x 4.6mm (Part No. 81404)Mobile Phase: 1%Ammonium Hydroxide in Methanol:Acetonitrile (25:75)Flow Rate: 2.0mL/minDetector: UV at 280nm

1. 2,3-MDA 2. 3,4-MDA 3. 2,3-MDMA 4. 3,4-MDMA

Column: Alltech® Alltima HP Silica, 3µm, 150 x 4.6mm (Part No. 87786)Mobile Phase: 1%Ammonium Hydroxide in Methanol:Acetonitrile (75:25)Flow Rate: 1.0mL/minColumn Temp: 25°CDetector: UV at 280nm

1. 2,3-MDA 2. 3,4-MDA 3. 2,3-MDMA 4. 3,4-MDMA

MDA & MDMA AnalogsMDA and MDMA Analogs

8384

1. Theobromine 2. Theophylline 3. 8-Chlorotheophylline 4. Caffeine 5. 7-(2-Chloroethyl)theophylline

0 1

1

2

3 4 5

2 Min.

1. Xanthine 2. Theobromine 3. Theophylline 4. Caffeine 5. 8-Chlorotheophylline

Column: Platinum™ C18-EPS 100Å, 3µm, 7 x 33mm Rocket™ Mobile Phase: 0.010M Sodium Acetate, pH 4.0:Methanol (70:30) Flow Rate: 5.0mL/min Detector: UV at 254nm

Rocket™ Columns for High Throughput

with Conventional LC

8679


® ®

mA

U

140

160

180

120

80

100

40

60

0

20

2 4 6 8 10 12 14 16 Min.0

10708

Nifedipine


Mobile Phase: 0.39% Sodium Phosphate Monobasic, pH4.5:

Acetonitrile (35:65)


Detector: UV

Statins

Simvastatin Tablets (U.S.P.)

at

ph

arm

ace

uti

ca

lshplc applications

10246

10262

10265

1. Amiloride HCl Hydrate 2. Acetazolamide 3. Hydrochlorothiazide 4. Triamterene 5. Chlorthalidone 6. Furosemide 7. Hydroflumethiazide 8. Clopamide 9. Ethacrynic Acid 10. Trichloromethiazide 11. Probenecid 12. Indapamide 13. Bumetanide 14. Bendroflumethiazide 15. Spironolactone

1. Amiloride HCl Hydrate 2. Acetazolamide 3. Hydrochlorothiazide 4. Chlorthalidone 5. Hydroflumethiazide 6. Clopamide 7. Trichloromethiazide 8. Indapamide 9. Bendroflumethiazide 10. Bumetanide 11. Probenecid 12. Spironolactone

1. Acetazolamide 2. Hydrochlorothiazide 3. Triamterene 4. Hydroflumethiazide 5. Clopamide 6. Trichloromethiazide 7. Indapamide 8. Bumetanide 9. Spironolactone

Column: Alltech® Alltima C18, 3µm, 53 x 7mm Rocket™ (Part No. 50605)

Mobile Phase: A: 0.1% Trifluoroacetic Acid in 0.025M Ammonium Acetate B: 0.1% Trifluoroacetic Acid in Acetonitrile

Gradient: Time: 0 4

%B: 20 95






%B: 25 95 95


Detector: ELSD




%B: 25 65



Diuretics Diuretics and Uricosuric Agents

Doping Agents

Diuretics — High Throughput


®

®

®

0 2

1

2

3 4

5

4 6 8 10 Min.

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.

1

2 34

0 1

1

2

2

3

3

4

4

5 6 7 8 9 Min.

0 1 2 3 4 5 Min.

1

2

pharm

aceutic

als

hplc applications

Column: Alltech® Platinum™ EPS C18, 5µm, 150 x 4.6mm

(Part No. 32043)




1. Sulfanilamide 2. Cefaclor 3. Cefatrizine 4. Thiamphenicol 5. Cefotaxime

1. Oxytetracycline Dihydrate 2. Tetracycline HCl 3. Meclocycline Sulfosalicylate Salt


Mobile Phase: A: 0.05M KH2PO4, pH3.0 B: Acetonitrile


%B: 15 40




Mobile Phase: 0.3% Pentafluoropropionic Acid in 43.7mM Ammonium

Formate, pH2.7:0.3% Pentafluoropropionic Acid in

Methanol (45:55)


Detector: ELSD

1. Streptomycin 2. Amikacin 3. Tobramycin 4. Neomycin




%B: 10 40 40


Detector: UV at 210

10319 10351


(Part No. 87697)

Mobile Phase: 0.1% Phosphoric Acid:Acetonitrile (85:15)



Column: Alltech® Prevail™ Select C18, 5µm, 150 x 4.6mm

(Part No. 99300)

Mobile Phase: Ammonium Hydroxide in Water, pH9.0:Acetonitrile (5:95)


Detector: ELSD

8711 8764

93969210

1. Tylosin (400ppm) 2. Monensin (180ppm)

1. Neomycin 2. Amoxicillin 3. Vancomycin 4. Cefatrizine 5. Metampicillin 6. Cefotaxime 7. Unknown 8. Bacitracin 9. Pipericillin 10. Bacampicillin 11. Oxacillin

Antibiotics

Antibacterials Antibacterials

Aminoglycoside Antibiotics Antibacterials

Sulfa DrugsTylosin and Monensin

1. Sulfanilamide 2. Sulfamerazine 3. Sulfamethazine 4. Sulfamethizole


®

®

®

1

2

3456 7

80

5

10

15

20

mA

U

0 1 2 3 4 5 Min.0 5 10 15 20 25 30 35 40 45 Min.

1

2

3

pharm

aceuticals

hplc applications

10724

1. Sulfadiazine 2. Sulfathiazole 3. Sulfamerazine 4. Sulfamethazine 5. Sulfamethoxazole 6. Sulfaisoxazole 7. Sulfaquinoxilone 8. Sulfamethoxine


(Part No. 5139606)

Mobile Phase: A: 0.1% Formic Acid B: Methanol

Gradient: Time: 0 1.4

%B: 20 100



10626

1. Amikacin 2. Tobramycin 3. Neomycin

Column: ® A ltima HP HILIC, 5µm, 150 x 4.6mm

(Part No. 86464)

Mobile Phase: Acetonitrile:Methanol:Water (87:11:2)


Detector: ELSD

10697

Column: C18, 5µm, 250 x 4.6mm

(Part No. 5138810)

Mobile Phase: Monobasic Potassium Phosphate, pH5.0:

Acetonitrile (96:4)

Flow Rate:

Detector: at 230nm

Sulfa DrugsAntibiotics via HILIC

Amoxicillin (U.S.P.)

Antibiotics

0 1 2 3 4 5 6 7 Min.

-500000

0

500000

1000000

1500000

2000000

2500000

3000000

3500000

4000000

4500000

5000000

5500000

6000000

mA

U

Neomycin Extracted from Neo-Cinolone Cream

Column C18 column, 5µm, 250 x 4.6mm

Mobile Phase: 0.3% Pentafluoropropionic Acid in Methanol:

0.3% Pentafluoropropionic Acid in 43.4mM

Ammonium Formate, pH 2.6 (55:45)

Flowrate: 1.0mL/min

Detector: ELSD

1. Neomycin

0 1

1

2 3 4 5 6 7 8 9 Min.

GraceSmart™

1.5mL/min

UV

0.05N

9216


®®

Alltech

® Alltima ® Alltech

0 1

1

2 3 4 5 6 7 8 9 Min.

ph

arm

ace

utic

als

hplc applications

10680

1. Spectinomycin


(Part No. 87829)

Mobile Phase: 10mM Ammonium Carbonate:Acetonitrile (98:2)


Detector: ELSD


(Part No. FM15960E)

Mobile Phase: Acetonitrile:0.1% Trifluoroacetic Acid (31:69)



Bacitracin SpectinomycinAntibiotics

mA

U

80

100

60

40

20

0

4 62 8 10 12 14 18 Min.160

10760

1. Clarithromycin


Mobile Phase: Phosphate Buffer (Dissolve 9.11g Potassium Dihydrogen

Phosphate in 1000mL Water, add 2mL Triethylamine, adjust

with Phosphoric Acid to pH of 5.5):Acetonitrile (60:40)



Clarithromycin on Chinese Pharmacopeia Assay

mA

U 800

1000

1200

1400

600

400

200

0

5 10 15 25 Min.200

10776

1. Ceftriaxone


Mobile Phase: 0.02mol/L n-Octylamine:Acetonitrile (73:27)



Ceftriaxone Sodium on Chinese Pharmacopeia Assay


®

, , ,

pharm

aceuticals

hplc applications

Bronchodilators


(Part No. FM15960E)

Mobile Phase: Acetonitrile:50mM Aqueous Ammonium Acetate, pH4.7

(12:88)



Benzoate

-Ethoxytheophylline

Xanthines

Clotrimazole

Lotrimin® Topical Assay

Antifungal

10447


Mobile Phase: A: Water to pH7.0 with Acetic Acid B: Methanol

Gradient: Time: 0 8 10 20 25

%B: 30 50 100 100 30


Detector: ELSD

Griseofulvin Assay on Chinese Pharmacopeia


Mobile Phase: 0.05mol/L Potassium Dihydrogen Phosphate:

Acetonitrile:Methanol (57:38:5)



1. Uracil 2. Theobromine 3. Impurity 4. Sodium 5. Theophylline 6. 7. Caffeine

0 2 4 6 8 10 12 14 16 18 20 22 24 Min.

2

1. PEG 2.

mA

U

300

350

400

250

150

200

100

50

0

12.510 17.5155 7.52.5 20 22.5 Min.0

1. Griseofulvin

10772


®

1

0 1

89

10 11

1213

1

2

2

3

3

4

4,5

5 6

6

7

7

8 Min.

pharm

aceutic

als

hplc applications

Column: Alltech® Alltima Phenyl 3µm, 53 x 7mm Rocket™

(Part No. 43867)

Mobile Phase: 0.05% Trifluoroacetic Acid:Methanol:Acetonitrile

(50:19:31)



1. 10-Deacetylbaccatin III 2. Baccatin III 3. 7-Xylosyl-10-deacetyltaxol B 4. Taxinine M 5. 7-Xylosyl-10-deacetyltaxol 6. 7-Xylosyl-10-deacetyltaxol C

9221

Taxol® and Related Taxanes in Yew

Taxols

7. 10-Deacetyltaxol 8. 7-Xylosyltaxol 9. Cephalomannine (Taxol® B) 10. 7-Epi-10-deacetyltaxol 11. Taxol®

12. Taxol® C 13. 7-Epitaxol

1. 10-Deacetylbaccatin III 2. Baccatin III 3. 10-Deacetyl-7-xylosyltaxol B 4. Taxinine M 5. 10-Deacetyl-7-xylosyltaxol 6. 10-Deacetyl-7-xylosyltaxol C 7. 10-Deacetyltaxol 8. 7-Xylosyltaxol

Taxanes

9. Cephalomannine 10. 10-Deacetyl-7-epitaxol 11. Taxol®

12. Taxol® C 13. 7-Epitaxol a. 3’-Fluorophenyl-10-deacetyltaxol b. 3’-Fluorophenyltaxol

Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 218MS54)

Mobile Phase: A: 50mM Sodium Acetate, pH6.8 B: Acetonitrile


%B: 34 34 44 60 100



mA

U

200

250

300

350

400

450

150

100

50

0

2 4 6 8 10 Min.0


Mobile Phase: Methanol:Water (10:90)



1. Aciclovir

Aciclovir Assay on Chinese Pharmacopeia

Antiviral

10764


®

mA

U

250

150

200

100

50

0

6 842 10 12 Min.0

mA

U

20

25

30

15

10

5

0

5 10 15 2520 30 35 Min.0

mA

U

120

140

100

60

80

40

20

0

6 842 10 12 Min.0

mA

U

200

250

300

150

100

50

0

2.5 5 7.5 10 12.5 15 17.5 20 22.5 25 Min.0

mA

U

600

700

500

300

400

200

100

0

6 842 1210 14 16 20 Min.180

ph

arm

ace

uti

ca

lshplc applications

Nimodipine Tablets on Chinese Pharmacopeia Assay


Mobile Phase: Methanol:Phosphate Buffer (Dissolve 1.36g Potassium

Dihydrogen Phosphate in 1000mL Water, add 4mL

Triethylamine, adjust with Phosphoric Acid to pH3.5)

(75:25)



1. Flunarizine 1. Nimodipine

10763

Flunarizine Hydrochloride Capsules on Chinese Pharmacopeia Assay

Calcium Channel Blockers

10761


Mobile Phase: Methanol:Acetonitrile:Water (35:38:27)




Mobile Phase: 700mL Phosphate Buffer (6.8g Potassium Dihydrogen

Phosphate in 1000mL Water, adjust pH3.0 with

Phosphoric Acid), add Methanol 300mL and Sodium

1-Octanesulfonate 1.30g



1. Atenolol

Atenolol Assay on Chinese PharmacopeiaBeta Blockers

10777


Mobile Phase: Sodium 1-Heptanesulfate solvent (2.0g Sodium

1-Heptanesulfate in 900mL Water, adjust to pH 3.9

with Acetic Acid):Acetonitrile:Methanol (25:6:1)



1. Famotidine 1. Naloxone

10778

Famotidine Tablets on Chinese Pharmacopeia Assay

Histamine Receptor Antagonists

10779


Mobile Phase: Sodium 1-octanesulfonate solvent (1.36g Sodium

1-octanesulfonate, 1.0g Sodium Chloride in 580mL

Water):Methanol:Phosphoric Acid (580:420:1)



Naloxone Hydrochloride Injection on Chinese Pharmacopeia Assay

Narcotic Analgesic Antagonists


500000

600000

200000

300000

400000

0

-100000

100000

70 80 906040

12

3

50 100 Min.0

1

2

3

4

5

67

8

9

10

11

20 40 60 Min.

1

2

3

4

0 2.5 5 7.5 10 12.5 15 17.5 20 22.5 25 Min.

hplc applicationsn

utra

ce

utic

als

| vita

min

s


(Part No. 35101)

Mobile Phase: A: Acetonitrile:Water:Isopropyl Alcohol (80:5:15)

B: Acetonitrile:Water:Isopropyl Alcohol (67:21:12)

Gradient: Time: 0 28 35 45 50 51 57

%B: 25 85 80 75 90 100 25


Detector: ELSD

1. Rh2 2. Rh1 3. Rg2 4. Rg3 5. Rg1 6. Rf

7. Re 8. Rd 9. Rc 10. Rb2 11. Rb1

10239 10287

10288

1. Bilobalide 2. Ginkgolide J 3. Ginkgolide C 4. Ginkgolide A 5. Ginkgolide B

1. Bilobalide 2. Ginkgolide J 3. Ginkgolide C 4. Ginkgolide A 5. Ginkgolide B

10142

1. Quercetin 2. Kaempferol 3. Isorhamnetin

Column: Alltech® Alltima C18, 3µm, 100 x 4.6mm

(Part No. 81382)


B: 0.1% Trifluoroacetic Acid in Methanol


%B: 10 55


Detector: ELSD


(Part No. 32064)

Mobile Phase: A: 0.05% Trifluoroacetic Acid in Water:Methanol (95:5)



%B: 10 90


Detector: ELSD


(Part No. 50605)



Gradient: Time: 0 5

%B: 10 45


Detector: ELSD

10742


Mobile Phase: Methanol:Tetrahydrofuran:Water (25:10:65)


Detector: ELSD

1. Ginkgolide C 2. Bilobalide 3. Ginkgolide A 4. Ginkgolide B




%B: 19 19 29 29 40


Detector: ELSD

10765

Ginseng

Ginsenosides in Korean Ginseng

Ginkgo Biloba

Ginkgo Biloba Flavonol Glycoside Standards Ginkgo Biloba Terpene Lactone Standards

Terpene Lactones High ThroughputFolium Ginkgo

Ginsenoside

1. Rg1 2. Re 3. Rb1


®

®

1

2

3

54

0 5 10 15 20 25 30 35 40 45 Min.

0 5

1

2

3

4

5 6

10 15 Min.

hplc applicationsn

utr

ace

uti

ca

ls | v

ita

min

s

10315

Column: Alltech® Alltima C18, 5µm, 150 x 10mm (Part No. 81102)




%B: 22 32


Detector: ELSD

1. Bilobalide

2. Ginkgolide J

3. Ginkgolide C

4. Ginkgolide A

5. Ginkgolide B




%B: 20 40 40


Detector: UV at 280

1. Myricetin 2. Quercetin 3. Naringenin 4. Hesperetin 5. Apigenin 6. Kaempferol

Column: Alltech® Alltima C18 3µm, 53 x 7mm Rocket™

(Part No. 50605)

Mobile Phase: 0.1% Trifluoroacetic Acid in Water:

0.1% Trifluoroacetic Acid in Acetonitrile (65:35)


Detector: ELSD

1. Myricetin 2. Quercetin 3. Apigenin 4. Kaempferol


Mobile Phase: A: 0.05M K2HPO4, pH2.5 B: Acetonitrile


%B: 20 40



9107 9110

1. Protocatechuic Acid 2. Chlorogenic Acid 3. Caffeic Acid


(Part No. 50605)

Mobile Phase: 0.01M K2HPO4, pH2.6:Acetonitrile (80:20)



1. Pseudohypericin 2. Hypericin


(Part No. 50605)

Mobile Phase: 0.2% Phosphoric Acid:Methanol (5:95)


Detector: Vis at 585nm

9397

9271 8887

1. Naringin 2. Neohesperidin 3. Myricetin 4. Quercetin 5. Naringenin 6. Kaempferol

Liquid Ginkgo Biloba Extract Prep

Flavonoids

Echinacea

Polyphenolic Flavonoids

Flavonoid Mix Flavonoid Mix

St. John’s Wort

Phenolic AcidsHypericins

Ginkgo Biloba


®

® ®

® ®

2

1

0 2 4 6 8 10 12 14 Min.

0 2 4 6 8 10 12 14 16 18 20 Min. 0 1 2 3 4 5 6 7 8 9 10 Min.

0 2 4 6 8 10 12 14 16 Min.

0 2 4 6 8 10 12 Min.

hplc applicationsn

utra

ce

utic

als

| vita

min

s

10747

1074610745

10744


Mobile Phase: Methanol:Water:Acetic Acid (87:13:0.1)


Detector: ELSD

10743

1. Peimine 2. Peininine


Mobile Phase: Methanol:Water:Acetic Acid (85:15:0.2)


Detector: ELSD


(Part No. 5138810)



Detector: ELSD


Mobile Phase: Acetonitrile:Water:Diethylamine (70:30:0.3)


Detector: ELSD




Detector: ELSD

1. Liguidanbaric Acid 1. Momordin lc

1. Sarsasapogenin

1. Astragaloside IV

Chinese Pharmacopeia

Fructus Liquidambaris Fructus Kochiae

Bulbus Fritillariae Thunbergii Rhizoma Anemarrhenae

Radix Astragali


mA

U

50

20

30

40

10

0

25 30 35 4015 205 10 45 Min.0

mA

U

500

200

300

400

0

100

15 20 25100 5 30 Min.

0 2

12

3

4 6 8 10 12 14 16 18 20 Min.

0 1

1

2

2

3

3

4

5

6

7 8

4

5 Min.

hplc applicationsn

utr

ace

uti

ca

ls | v

ita

min

s


Mobile Phase: 0.05% Trifluoroacetic Acid:Acetonitrile (87:13)

Flow Rate: 23mL/min

Detector: ELSD

1. Caffeine 2. Epicatechin 3. Epigallocatechin Gallate




Detector: ELSD

10139

1. Caffeine 2. Epicatechin 3. Epigallocatechin Gallate

Column: Alltech® Platinum™ EPS C18,

1.5µm, 33 x 7mm Rocket™ (Part No. 50577)




1. Gallic Acid 2. Epigallocatechin 3. Catechin 4. Epicatechin 5. Epigallocatechin Gallate 6. Caffeine 7. Gallocatechin 8. Epicatechin Gallate

10140

8825

Green Tea

Green Tea Extract - PreparativeGreen Tea Extract - Analytical

Catechins

1. Gallic Acid 2. Gallocatechin 3. Epigallocatechin 4. Catechin 5. Epicatechin 6. Epigallocatechin Gallate 7. Gallocatechin Gallate 8. Epicatechin Gallate 9. Catechin Gallate

Catechins and Gallic Acid in Green Tea


Mobile Phase: Methanol:0.1% Citric Acid (25:75)



10773

1. Puerarin

Puerarin

Puerarin





10769

1. Tanshinone II

Tanshinone II

Tanshinone II


Mobile Phase: A: Acetonitrile:25mM Phosphoric Acid (10:90)

B: Acetonitrile:25mM Phosphoric Acid (90:10)


B%: 0 0 15




® ®

Min.

0 1 2 3 4 5 6 7 8 9 10 Min.

1

2

3

2

3

4

1

5

6

7

0 2 4 6 8 10 12 14 Min.

0 2 4 6 8 10 12 14 16 Min.

5 6

71

2

43

0 2 4 6 8 10 12 14 Min.

1

2

3

4

5

6

7

hplc applicationsn

utra

ce

utic

als

| vita

min

s

1. Thiamine 2. Ascorbic Acid 3. Nicotinic Acid 4. Niacinamide 5. Pantothenic Acid 6. Riboflavin 7. Cyanocobalamin 8. Biotin 9. Vitamin A 10. -Tocopherol 11. -Tocopherol 12. -Tocopherol 13. Vitamin D2 14. Vitamin D3 15. Vitamin K

9545

1. Vitamin A (100ng on column) 2. Vitamin E (160ng on column) 3. Vitamin K1 (130ng on column)


Mobile Phase: Acetonitrile:Methanol (75:25)



1. Vitamin A 2. -Tocopherol 3. -Tocopherol 4. -Tocopherol (Vitamin E) 5. Vitamin D2 6. Vitamin D3 7. Vitamin K




Detector: UV at 212

1. Thiamine 2. Ascorbic Acid 3. Unknown 4. Fumaric Acid 5. Pyridoxine 6. Niacinamide 7. Pantothenic Acid

1. Ascorbic Acid 2. Nicotinic Acid 3. Pyridoxamine 4. Niacinamide 5. Folic Acid 6. Riboflavin 7. Vitamin B12


(Part No. 32043)

Mobile Phase: A: 0.1M Ammonium Phosphate, pH4.5

B: Acetonitrile


B%: 5 5 40 40




Mobile Phase: Methanol:Chloroform (90:10)


Detector: ELSD

9390

81549456


Mobile Phase: A: Water pH3.2 w/Formic Acid

B: Acetonitrile:Methanol (83:17)

Gradient: Time: 0 3 10 10.1 30

%B: 0 0 45 100 100


Detector: ELSD

10279

1. Ascorbic Acid 2. Nicotinic Acid 3. Pyridoxamine 4. Niacinamide 5. Folic Acid 6. Vitamin B12 7. Riboflavin


Mobile Phase: A: 0.1M Ammonium Dihydrogen Orthophosphate

Buffer, pH4.5

B: Acetonitrile


B%: 5 5 40 40



10733

Fat-Soluble Vitamins

Water- and Fat-Soluble Vitamins

Fat-Soluble Vitamins and Tocopherols

Water-Soluble VitaminsWater-Soluble Vitamins

from Multi-Vitamin Tablet

Water-Soluble Vitamins

Vitamins


®

Column: VYDAC 201TP54 (C18, 5 µm, 4.6 mm i.d. x 250 mm)

hplc applicationsn

utr

ace

uti

ca

ls | v

ita

min

s




Detector: ELSD

1. -Tocopherol 2. -Tocopherol 3. -Tocopherol

9458

Vitamin E Soft Gel Tablet

1. All-trans-Retinol 2. Tocol 3. -Tocopherol 4. -Tocopherol 5. Lutein 6. Zeaxanthin 7. Cryptoxanthin 8. -Carotene 9. All-trans- -Carotene 10. cis- -Carotene

Determination of Retinol, -Tocopherol, and -Carotene in Serum

Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 201TP54)

Mobile Phase: A: Buffer:Methanol:n-Butanol (15:75:10)

B: Buffer:Methanol:n-Butanol (2:88:10)

Buffer:Aqueous 0.02M Ammonium Acetate, pH3.5


%B: 0 0 100 100


Detector: Absorbance, programmed wavelength


Mobile Phase: Methanol:Ethanol (50:50)



10771

Coenzyme Q10

mA

U

80

100

60

40

20

0

15 20105 25 30 35 45 Min.400

1. Coenzyme Q10


Mobile Phase: Methanol:Chloroform (94:6)

–Carotenes

Detector: UV Flow Rate: 1.0mL/min

1. -Carotene 2. cis- -Carotene 3. -Carotene 4. 13-cis- -Carotene 5. 9-cis- -Carotene

1. Vitamin D2

2. Vitamin D3

Column: Adsorbosphere™ UHS C18, 5µm, 4.6 x 150mm

(Part No. 288118)

Mobile Phase: 80% Acetonitrile 20% Methanol



Column Temp: Ambient

Vitamins D2 & D3

5401

Vitamins


0 10 20 30 40 Min.

1

2

45

67

8

1

1

UV

RI

ELSD

0 1 2 3 4 5 Min.


Mobile Phase: A: Acetonitrile B: Dichloromethane


%B: 30 50


Detector: ELSD

1. LGG 2. LGL 3. LLL 4. OGL 5. PGL 6. OLL 7. OGO + PLL 8. OOO

10047

Evening Primrose

Triglycerides

10412


(Part No. 35102)



Detector: ELSD

10413

10411

Glucosamine

1. Glucosamine


Mobile Phase: A: 1% Formic Acid B: Acetonitrile


%B: 20 100 100 20 20


Detector: ELSD

1. Cimiracemoside A 2. Actein 3. 27-deoxyactein

9496

Black Cohosh Extract

Black Cohosh


(Part No. 50605)

Mobile Phase: 0.02M KH2PO4, pH2.6:Methanol (52:48)



1. Hydrastine 2. Berberine

9124

Alkaloids

Goldenseal Root

Glucosamine


(Part No. 50605)

Mobile Phase: 0.05% Trifluoroacetic Acid:Methanol:Dioxane (75:23:2)



9191

Iridoid Compounds

Devil’s Claw

1. 8-para-Coumaroyl Harpagoside 2. Harpagoside

0 1

1

2

2 3 4 5 6 Min.

Column:

Mobile Phase: Dioxane:ACN:0.05% TFA (3:64:33)Flowrate: 3mL/minDetector: UV at 220nm

1. Acetoxyvalerenic Acid2. Valerenic Acid

Valerian

Valerian

9160

® Alltima C18, 3µm, 53 x 7mm Rocket™

(Part No. 50605) Alltech


nu

trace

utic

als

| vita

min

shplc applications

®

®

® ®

Carbohydrates


hplc applicationsfo

od

& b

eve

rag

e

0 10 20 30 40 50 60 Min.

1

2

3

45 6

7

8

910

11

1213

14

15

16

17

18

1920

21

22

0 1 2 3 4 Min.

1

2

3

Column: Alltech® Prevail™ Carbohydrate ES,

5µm, 53 x 7mm Rocket™ (Part No. 35104)

Mobile Phase: A: Acetonitrile B: Water

Gradient: Time: 0 4

%B: 32 50


Detector: ELSD

Column: Alltech® Prevail™ Carbohydrate ES, 5µm,

250 x 4.6mm (Part No. 35101)

Mobile Phase: A: Acetonitrile B: 0.04% Ammonium Hydroxide in Water


%B: 17 27 45 55


Detector: ELSD

9420 1. Dextrose 2. Maltose 3. Maltotriose 4. Maltotetraose 5. Maltopentaose 6. Maltohexaose 7. Maltoheptaose 8. Maltooctaose

10009

Column: Alltech® Prevail™ Carbohydrate ES, 250 x 4.6mm

(Part No. 35101)



%B: 20 35


Detector: ELSD

1. iso-Erythritol 2. Fructose 3. Sorbitol 4. Mannitol 5. Glucose 6. Inositol 7. Sucrose 8. Maltitol 9. Maltose


(Part No. 35101)

Mobile Phase: A: Acetonitrile:Acetone (75:25)

B: Water


%B: 24 50


Detector: ELSD

10. Raffinose 11. DP3 12. DP4 13. DP5 14. DP6 15. DP7 16. DP8 17. DP9 18. DP10 19. DP11 20. DP12 21. DP13 22. DP14

Column: Alltech® Prevail™ Carbohydrate ES,

5µm, 53 x 7mm Rocket™ (Part No. 35104)



Column Temp: 30°C

Detector: ELSD

10277


(Part No. 35101)



%B: 35 50


Detector: ELSD

1. Fructose 2. Glucose 3. Sucrose 4. Maltose 5. Maltotriose 6. Maltotetraose 7. Maltopentaose

1. Fructose 2. Glucose 3. Sucrose 4. Maltose 5. Maltotriose 6. Maltotetraose 7. Maltopentaose

9350 9360

1. Fructose 2. Glucose 3. Sucrose

1. Dextrose 2. Maltose 3. Maltotriose 4. Maltotetraose 5. Maltopentaose

6. Maltohexaose 7. Maltoheptaose 8. Maltooctaose 9. Maltononaose 10. Maltodecaose 11. Maltoundecaose 12. Maltododecaose 13. Maltotridecaose

9354

Corn Syrup SolidsCorn Syrup Oligomers

Mono-, Di-, Tri- Saccharides Sugars

Beer Standards

Unmodified Mobile Phase

Beer Standards

Modified Mobile Phase

0 5 10 15 20 25 30 35 40 45 Min.

1

23

4

0

1

2

3 5

7

8 9

10

4

6

10 20 30 40 50 Min.

0 1 2 3 Min.

1

2

3

4

5


(Part No. 35101)



Detector: ELSD

1. iso-Erythritol 2. Xylitol 3. Mannitol 4. Maltitol


(Part No. 35101)



%B: 20 35


Detector: ELSD

9351

9474

Column: Alltech® IOA-1000, 9mm, 300 x 7.8mm (Part No. 9646)

Mobile Phase: 0.005N Sulfuric Acid


Column Temp: 60°C

Detector: RI

5411

9446

9352


Mobile Phase: Acetonitrile:0.05M Ammonium Acetate, pH5.4, (20:80)


Detector: UV at 230

1. Ascorbic Acid 2. Saccharin 3. Benzoic Acid 4. Aspartame 5. Vanillin


(Part No. 35101)



%B: 17 35


Detector: ELSD

1. Fructose 2. Sorbitol 3. Glucose 4. Maltose

1. Fructose 2. Glucose 3. Maltose 4. Maltotriose 5. Maltopentaose 6. Maltohexaose 7. Maltoheptaose

1. Citric Acid 2. Tartaric Acid 3. Glucose 4. Malic Acid 5. Fructose 6. Acetic Acid 7. Glycerol 8. Lactic Acid 9. Methanol 10. Ethanol

Sugar Alcohols Wine Analysis - 2000 Sipaska Rosé

Wine Analysis by Ion Exclusion

Beverage AdditivesImported Ale


Gradient: 7 to 10% Methanol over 60 min.,

then to 25% Methanol at 90 min


Detector: ELSD

Partial Digest: Complex Polysaccharide

from Plant Cell Wall

Carbohydrates


foo

d &

be

ve

rag

ehplc applications

0 2 4 6 8 10 12 14 16 18 Min.

2 4

53

1

0 5 10 15 20 25 30 35 40 Min.

12

3 4

0 5 10 15 20 25 30 35 40 Min.

1

87

65

4

32

0 5 10 15 20 25 Min.

2

1

3

0 5 10 15 20 25 30 35 40 Min.

2

1

0 1 2 3 4 5 6 7 8 9 Min.

1

2 3

10531 1. Sucralose 2. Glucose 3. Maltose 4. Maltodextrins

10530

1. Sucralose 2. Unknown 1 3. Fructose 4. Glucose 5. Unknown 2 6. Sucrose 7. Maltose 8. Maltodextrins

10529

1. Fructose 2. Glucose 3. Sucrose

10579

1. Sucrose 2. Lactose


(Part No. 35101)



Detector: ELSD


(Part No. 35101)



%B: 20 29


Detector: ELSD

1. Fructose 2. Mannitol 3. Glucose 4. Sucrose 5. Unknown

10585


(Part No. 35101)



%B: 20 20 30 30 50


Detector: ELSD


(Part No. 35101)



%B: 20 20 30 30 50


Detector: ELSD


(Part No. 35101)



%B: 20 20 30 30 50


Detector: ELSD

10584

1. Sucralose 2. Rhamnose 3. Cellobiose


(Part No. 35102)



Detector: ELSD

Spiked UrineChromatogram courtesy of Rosemary Rose,

Monash University, Dept. of Gastroenterology

Conquistador® Tequila Splenda® Sweetener

Pepsi One® Cola Infant Formula

Flavor Sample SolutionChromatogram courtesy of Charles Stewart, Firmenich Inc.

Carbohydrates


foo

d &

be

ve

rag

ehplc applications

Min. Min.

Min. Min.

Min. Min.

0 5 10 15 20 25 30 35 40 Min.

1

2

3

4

5 6

0 5 10 15 20 25 30 35 40 Min.

4

3

1 2 5

45

3

21

1/3 Less Sugar

Regular

0 5 10 15 20 25 30 35 40 Min.

1

3

2

4

Kellogg's Frosted Flakes® Cereal

Regular vs. 1/3 less sugar

1. Sucralose 2. Fructose 3. Sorbitol 4. Glucose 5. Sucrose 6. Maltose

10526

10557

1. Fructose 2. Glucose 3. Sucrose 4. Maltose 5. Oligosaccharides

1. Fructose 2. Glucose 3. Sucrose 4. Unknown


(Part No. 35101)



%B: 20 20 30 30 50


Detector: ELSD


(Part No. 35101)



%B: 20 20 30 30 50


Detector: ELSD


(Part No. 35101)



%B: 20 20 30 30 50


Detector: ELSD


(Part No. 35101)



%B: 15 15 25 25 50


Detector: ELSD

1. Fructose 2. Glucose 3. Sucrose 4. Maltose

10556

10527

10528

Carbohydrates

Robitussin® Flu Syrup

Tobacco

Cough Syrup

Vegetable Fermentation Broth Standards

1. iso-erythritol

2. Fructose

3. Sorbitol

4. Mannitol

5. Glucose

6. Inositol

7. Sucrose

8. Maltitol

9. Maltose

10. Raffinose

11. MaltotrioseColumn: Prevail™ Carbohydrate ES Column, 250 x 4.6mm, (Part No. 35101)Mobile Phase: A: Acetonitrile B: 0.04% NH

4OH

Gradient: Time: 0 60 %B: 17 35Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient

0 10 20 30 40 50 Min.

12

3

4

5

6

7

8

9

10

11

10004


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ehplc applications

Carbohydrates


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ehplc applications

Sugar Alcohols

Column: Alltech® Prevail™ Carbohydrate ES column, 5µm,

250 x 4.6mm, (Part No. 35101)


Flow Rate: 1.0 mL/min

Column Temp: 30°C

10627

10635

18 Min.

2 31

ELSD

RI

Solvent

Front

0 2 4 6 8 10 12 14 16

32

1

1. Xylitol (200ppm)

2. Sorbitol (200ppm)

3. Dulcitol (200ppm)

Orange Juice Profiles

9532

9531

9530

Column: Prevail™ Carbohydrate ES Column, 5µm, 53 x 7.0mm Rocket™ (Part No. 35104) Mobile Phase: Acetonitrile:Water (75:25)Flow Rate: 1.3mL/min Detector: ELSDColumn Temp: Ambient

0 1 2 3 4 5 6 7 8 Min.

1

2

Orange Drink

1

2

3

Dominick’s® 100%

1

2

3

Minute Maid® 100%

1. Fructose

2. Glucose

3. Sucrose

Carbohydrate Analysis with ELSD Improves Baseline Stability and Detection Sensitivity

Compared to RI and Low-Wavelength UV

Column: Prevail™ Carbohydrate ES Column, 250 x 4.6mmMobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 20 25 %B: 20 50 50Flow Rate: 1.0mL/minColumn Temp: Ambient

9355

9380

20 4 6 8 10 12 14 16 18 20 22 Min.

12

34

5 6UV at 195nm

1 2

3

4

56

ELSDRI

0 1 2 3 4 Min.

1

23

1 2 3

Solvent

Front

ELSD

Column: Prevail™ Carbohydrate ES Column, 5µm, 53 x 7.0mm Rocket™ (Part No. 35104) Mobile Phase: Acetonitrile:Water (75:25)Flow Rate: 2.0mL/min Column Temp: 30°C

10278

10277

1. Mannitol

2. Fructose

3. Glucose

4. Sucrose

5. Raffinose

6. Stachyose

1. Fructose

2. Glucose

3. Sucrose

30

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Wine - Concord Grape

Column: Prevail™ Carbohydrate ES Column, 5µm, 250 x 4.6mm (Part No. 35101)Mobile Phase: A: Acetonitrile B:WaterGradient: Time: 0 60 %B: 17 35Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient

1. Sorbitol

2. Fructose

3. Maltose

4. Inositol

10 15 20 25 30 35 40 45 Min.

12

4

Low Level Detection - 10ng on Column

1. Fructose

2. Sucrose

3. Raffinose

Column: Prevail™ Carbohydrate ES Column, 5µm, 150 x 4.6mm, (Part No. 35102) Mobile Phase: A: Water B: Acetonitrile Gradient: Time: 0 10 11 14 %B: 80 50 80 80Flow Rate: 1.0 mL/min Detector: ELSDColumn Temp: Ambient

0 1 2 3 4 5 6 7 8 9 10 Min.

2

1 3

Column: 250 x 4.6mm as identifiedMobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 15 %B: 35 50Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient

Enhance Detector Sensitivity With Stable Gradient Baselines

Carbohydrate ES

1. Dextrose

2. Maltose

3. Maltotriose

4. Maltotetraose

5. Maltopentaose

6. Maltohexaose

7. Maltoheptaose

8. Maltooctaose

9. Maltononaose

10. Maltodecaose

11. Maltoundecaose

12. Maltododecaose

13. Maltotridecaose

14. Maltotetradecaose

15. Maltopentadecaose

12 3

47

5 6

0 5 10 15 20 25 Min.

8 9

1

2

3

4 56

71011 1312

18

12

1314

15

19 20 21 22 23 24 Min.

9354

9379

9486

Prevail™

Column

Silica Based Amino Column

Carbohydrates

9476 10498

31

hplc applications


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Clover Honey

Column: Prevail™ Carbohydrate ES Column, 5µm, 250 x 4.6mm, (Part No. 35101)Mobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 15

%B: 25 40

Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient

1. Fructose

2. Glucose

3. Sucrose

0 2

2

1

4

3

6 8 10 12 14 16 18 20 Min.

1. Glucose (DP1)

2. DP3 (1,6-alpha; 1,4-alpha)

3. DP6 (1,6-alpha; 1,4-alpha)

4. DP13

5. DP18

Column: Prevail™ Carbohydrate ES Column, 5µm 250 x 4.6mm (Part No. 35101)Mobile Phase: A: Acetonitrile B: WaterGradient: Time: 0 50 60 %B: 35 60 60Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient

Glucooligosaccharides in Maltodextrin

0 6 12 18 24 30 36 Min.

1

23

4 5

21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 Min.

4

5

9535

0 5

1

2

3

4

5

10 15 Min.

1. Unknown 2. Sucrose 3. Glucose 4. Fructose 5. Sorbitol

Column: Alltech® Carbohydrate Cation, 10µm, 6.5 x 300mm

(Part No. 70057)

Mobile Phase: Water


Column Temp: 90°C

Detector: Alltech ELSD

100% Pure Red Delicious Apple Juice

(a 1:250 Dilution)

Carbohydrates

9534

9365 8965

Column: Prevail™ Carbohydrate ES Column, 5µm, 250 x 4.6mm, (Part No. 35101)Mobile Phase: A: Acetonitrile B: WaterGradient: Step (5 min. hold at 40% B, 5% B steps every 5 min. to 70% B)Flow Rate: 1.0mL/minDetector: ELSDColumn Temp: Ambient

Partially Hydrolyzed Dextran

0 5 10 15 20 25 30 35 40 Min.

0 5 10 15 20 25 Min.

1

2

3

45

67

8

910

11

0 1 2 3 4 5 6 7 8 Min.

1

2

3

4

5

0 2 4 6 8 10 Min.

Column: Alltech® Prevail™ Organic Acid, 5µm, 150 x 4.6mm

(Part No. 88640)

Mobile Phase: 0.025M KH2PO4, pH2.5



1. Oxalic Acid 2. Tartaric Acid 3. Formic Acid 4. Malic Acid 5. Lactic Acid 6. Acetic Acid 7. Diglycolic Acid 8. Maleic Acid 9. Citric Acid 10. Succinic Acid 11. Fumaric Acid

9384


(Part No. 88640)

Mobile Phase: 0.025M KH2PO4, pH2.5



1. Tartaric Acid 2. Malic Acid 3. Lactic Acid 4. Citric Acid 5. Fumaric Acid

Organic AcidsOrganic Acid Mix Rye Bread Extract

9484

Carbohydrates

9540

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ehplc applications


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2

3

4

5

6

7

8 9 10

1

UV

0

3

4

5

6

7

8

9

10

2 4 6 8 10 12 14 16 18 Min.

ELSD

Column: ® Alltima HP C18 HiLoad, 3µm, 150 x 4.6mm

(Part No. 87686)




%B: 80 100 100


7. Linoleic Acid 8. Palmitic Acid 9. Oleic Acid 10. Stearic Acid

1. Caproic Acid 2. Caprylic Acid 3. Capric Acid 4. Lauric Acid 5. Linolenic Acid 6. Myristic Acid

Organic Acids

Fatty Acids Using UV and ELSD in Tandem

10430


(Part No. 88640)



Detector: ELSD

1. Lauric Acid 2. Myristic Acid 3. Palmitic Acid 4. Stearic Acid 5. Arachidic Acid 6. Behenic Acid

9539

Underivatized Fatty Acids


(Part No. 32420)




1. Tartaric Acid 2. Malic Acid 3. Lactic Acid 4. Diglycolic Acid 5. Maleic Acid 6. Succinic Acid 7. Fumaric Acid

Organic Acid Mix

8244

Column:

Mobile Phase: THF:CH 3 CN:0.1% H 3PO4 (25:55:20)

Flowrate: 1.5mL/min


1. Capric Acid (C10)

2. Lauric Acid (C12)

3. Myristic Acid (C14)

4. Palmitic Acid (C16)

5. Stearic Acid (C18)

Fatty Acids

0 2 4 6 8 10 12 Min.

1

2

3

4

5

1

2

3

4

5

6

7

8

9

10

11

12

0 10 20 30 Min.

1. Oxalic Acid 2. Citric Acid 3. Shikimic Acid 4. Fumaric Acid 5. Butyric Acid 6. Homoprotocatechuic Acid 7. Gallic Acid 8. Protocatechuic Acid 9. Gentisic Acid 10. p-Hydroxybenzoic Acid 11. Benzoic Acid 12. Salicylic Acid

7067

Short-Chain Aromatic Carboxylic Acids

Column: Alltech® OA-2000, 6.5µm, 100 x 6.5mm (Part No. 9048)


Column Temp: 45°C



8060

1. Capric Acid 2. Lauric Acid 3. Myristic Acid 4. Palmitic Acid 5. Stearic Acid

9459

Long Chain Fatty Acids

0 1 2 3 4 Min.

1

2

3

4

5

Column: Alltech® Prevail™ C18, 3µm, 53 x 7mm Rocket™

(Part No. 99279)



Detector: ELSD

Alltech

® Alltima C18, 5µm, 150 x 4.6mm Alltech ®

®

1

2 3

4

5

0 1 2 3 4 5 6 7 Min.

14

13

12

11

10

98

4

56

7123

0 5 10 15 20 25 30 Min.

6

4

3

7

8

52

1

0 5 10 15 20 Min.




Detector: UV at 220

1. Tartaric Acid 2. Malic Acid 3. Lactic Acid 4. Maleic Acid 5. Fumaric Acid

10705

1. LLO 2. OOLn 3. PLnO 4. LOO 5. PoOO 6. PLO 7. PoOP 8. OOO 9. POO+SOL 10. POP 11. SOO 12. POS+SLS 13. PPS 14. C17

10561

Column: Alltech® Prevail™ Silica, 3µm, 150 x 3.0mm (Part No. 99341)

Mobile Phase: A: Chloroform; B: Methanol; C: Triethylamine buffer pH3


%B: 12 60 12

%C: 0.5 12 0.5


Detector: ELSD

Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm

(Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD

Olive Oil

1. LACCER 2. PA 3. PE 4. PI 5. PS 6. PC 7. SM1 8. SM2

10576

Acid Whey

Organic AcidsOrganic Acids

0 2 4 6 8 10 12 14 16 Min.

12

3

4

5

6

7

89

1. LLO 2. LLP 3. OOL 4. POL 5. PPL 6. OOO 7. OOP 8. PPO 9. OOS


(Part No. 81387)

Mobile Phase: A: Dichloromethane B: Acetonitrile


%B: 70 55 70 70


Detector: ELSD

9515

Oils / LipidsOlive Oil

70 Min.0 10 20 30 40 50 60

ELSD

UV at 254nm

10656

Column: Alltech® Alltima

(Part No. 43852)

Mobile Phase: A: 0.1% Formic Acid B: Methanol


%B: 60 100


Column Temp: 30°C

Plant Oil Extract

10657


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® C18, 3µm, 2.1 x 150mm®

®

Oils / Lipids


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0 2 4 6 8 10 12 14 16 Min.

1

2

3

4

5

6

7

8

910

0 2 4 6 8 10 12 14 16 Min.

1

2

3

4

56

0 5 10 15 20 25 30 35 Min.

1

2

3

4

5

6

78

9

10

0 10 20 30 40 Min.

1

2

3

45

67

8

13

4

56

7

8

910

11

12

14

132

0 5 10 15 20 25 30 35 Min.

64

3

5

2

1

0 5 10 15 20 Min.

Column: ® Alltima C18, 3µm, 150 x 4.6mm (Part No. 81387)



%B: 70 55 70 70


Detector: ELSD




%B: 70 55 70 70


Detector: ELSD

1. LLL 2. LLO 3. LLP 4. OOL 5. POL 6. PPL 7. OOO 8. OOP 9. PPO 10. OOS

1. Trilaurin (LaLaLa) 2. Trilinolein (LLL) 3. 1,2-Dilinoleyl-3-oleyl-rac-glycerol (LLO) 4. 1,2-Dilinoleyl-3-palmitin-rac-glycerol (LLP) 5. 1-Palmitin-2-olein-3-Linolein rac-glycerol (POL) 6. 1,2-Dipalmytyl-3-linolein-rac glycerol (PPL)


(Part No. 87610)

Mobile Phase: A: Acetonitrile B: Dichlor omethane


%B: 30 51 70 70 30


Detector: ELSD

9522

9517




%B: 30 50


Detector: ELSD

Column: Alltech® Prevail™ Silica, 3µm, 150 x 3.0mm

(Part No. 99341)

Mobile Phase: A: Chloroform

B: Methanol

C: Triethylamine buffer pH3


%B: 12 60 12

%C: 0.5 12 0.5


Detector: ELSD

1. LLL 2. LLO 3. LLP 4. OOL 5. POL 6. PPL 7. OOO 8. OOP 9. PPO 10. OOS

10581

Column: HP C18 Hi-Load, 5µm, 250 x 4.6mm

(Part No. 87698)



%B: 30 51 70 70 30


Detector: ELSD

1. LGG 2. LGL 3. LLL 4. OGL 5. PGL 6. OLL 7. OGO + PLL 8. OOO

10047

Evening Primrose Oil

10566

1. C11 2. LLLn 3. LLL 4. LLO 5. PLL 6. LOO 7. PLO

8. PLP 9. OOO 10. SLO 11. POO 12. POP 13. SOO/PPP 14 C17

1. PA 2. PE 3. PI 4. LPE 5. PC 6. LPC

10562

Lecithin, 0.5mg/mL

Sesame Seed Oil

Walnut Oil

Sesame Seed Oil

Corn Oil

®

® ®

Alltech ® Alltima®Alltech

®

1

34

5

6

7

8

9

1011

12

1415 16

17

13

2

0 5 10 15 20 25 30 Min.

0 5 10 15 20 25 30 Min.

1

3

4567

8 910

11

2

1

2

3

4 5

6

7

8

9

10

1112

13

0 5 10 15 20 25 30 35 Min.

0 5 10 15 20 25 30 Min.

0 5 10 15 20 25 30 35 Min.

6

1

23 45 7

8

9

11

14

131210

0 5 10 15 20 25 30 35 Min.

1

3

4

56

7

89

2

10573

1. C13 2. PLO 3. PLP 4. POO 5. PLS 6. POP 7. SOO/PPP

10563

1056510564

Column: Alltech® Alltima HP C18 Hi-Load, 3µm,

150 x 3.0mm (Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD


150 x 3.0mm (Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD


(Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD


(Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD

8. SLS 9. POS 10. PPS 11. SOS 12. PSS 13. SOA 14. C19

1. CCLa 2. C11 3. CLaLa 4. CCM 5. LaLaLa 6. CLaM

7. LaLaM 8. LaLaO 9. LaMM 10. LaMO 11. LaMP/MMM 12. MMP/LaPP/MPO 13. C17

1. CCLa 2. C11 3. CLaLa 4. CCM 5. LaLaLa 6. CLaM 7. LaLaM 8. LaLaO 9. LaMM

10. LaMO 11. LaMP/MMM 12. MMO/LaPO 13. MMP/LaPP/MPO 14. PLP 15. POO 16. POP 17. C17

1. C11 2. LLL 3. LLO 4. PLL 5. LOO 6. LLS 7. PLO 8. OOO 9. SLO 10. POO 11. C17

10569

1. LLL 2. LLO 3. PLL 4. LOO 5. LLS 6. PLO 7. OOO 8. SLO 9. POO

10567


150 x 3.0mm (Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD


150 x 3.0mm (Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD

Refined Sunflower OilSunflower Oil

Palm Kernel Olein Fraction Butter

Coconut Oil Cocoa Butter

Oils / Lipids


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® ®

® ®

Oils / Lipids


hplc applications

6

4

3

52

1

0 5 10 15 20 Min.

0 5 10 15 20 25 30 35 Min.

1

3 4

56

78

9

10

11

12

1415

16

13

2

0 5 10 15 20 25 30 Min.

134

5

6

7

8

9

10

11 1213

2

0 5 10 15 20 25 30 35 Min.

1

3

4

5

6

7

8

9

101112

14

15

1716

18

19

132

6

4

3

7

85

21

0 5 10 15 20 Min.

6

4

3

7 8

910

5

21

0 5 10 15 20 Min.

1057210571

1. C13 2. OLL 3. PLL 4. LOO 5. PLO 6. PLP 7. MOP 8. OOO

9. POO 10. PLS 11. POP 12. SOO/PPP 13. POS 14. PPS 15. SOS 16. C19


150 x 3.0mm (Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD

Column: Alltech® Alltima HP C18 Hi-Load, 3µm, 150 x 3.0mm (Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD


(Part No. 87610)



%B: 30 51 70 70 30


Detector: ELSD

1. CCLa 2. C11 3. CLaLa 4. CMM 5. LaLaLa 6. LaLaM 7. LaLaO

8. LaMM 9. LaMO 10. LaMP/MMM 11. MMO/LaPO 12. MMP/LaPP/MPO 13. C17


Mobile Phase: A: Chloroform B: Methanol; C: Triethylamine buffer pH3


%B: 12 60 12

%C: 0.5 12 0.5


Detector: ELSD

10570


Mobile Phase: A: Chloroform B: Methanol C: Triethylamine buffer pH3


%B: 12 60 12

%C: 0.5 12 0.5


Detector: ELSD

1. C11 2. LLnLn 3. LLLn 4. LLL 5. OLnL 6. PLLn 7. LLO 8. OOLn 9. PLL 10. LOO

11. LLS 12. PLO 13. PLP 14. OOO 15. SLO 16. POO 17. POP 18. SOO/PPP 19. C17

1. PE 2. PI 3. PS 4. PC 5. SM1 6. SM2

Butter

10578

1. GLUCCER 2. LACCER 3. PE 4. PI 5. PS 6. PC 7. SM1 8. SM2


Mobile Phase: A: Chloroform B: Methanol C: Triethylamine buffer pH3


%B: 12 60 12

%C: 0.5 12 0.5


Detector: ELSD

10575

Polar Lipid Standards

Palm Oil Palm Kernel Stearin Fraction

Soybean Oil

1. LACCER 2. U 3. PA 4. PE 5. PI 6. PS 7. PC 8. SM1 9. SM2 10. LPC

10577

Gouda Cheese

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1

0 2.5 5 7.5 10 12.5 15 17.5 20 Min.

0

0.5

1.0

1.5

2.0

mV(x100)

HPLC Method for Melamine

1

0 1 2 3 4 5 Min.

-1

1

2

3

mAU

0

4

5

UHPLC Method for Malamine

0 2.5 7.5 10 15 17.55 12.5 20 Min.

0

0.5

1.0

1.5

2.0

mV (x100)

UHPLC

HPLC

Melamine - HPLC Method Transfer to UHPLC

HILIC Column

Flow

Rate

Time

(min) Conc.

Inj.

Range

Loading

Range

HPLC®

UHPLC®

Melamine

0 2.5 7.5 10 15 17.55 12.5 20 Min.

0

100

200

300

mV

Melamine - ELSD Method

HPLC Column: ® 86466)Detector: ® 5135834)Mobile Phase:

Flow Rate:

Detection:

Column Temp:

ELSD 3300:

Injection:

Egg Yolk Phospholipids

Column: Allsphere™ Silica column, 3µm,

100 x 4.6mm

Mobile Phase: A: Isopropanol: Hexane: Water (58:40:2)

B: Isopropanol: Hexane: Water(52:40:8)


%B: 0 100 100 0


Detector: ELSD

1. Phosphatidylethanolamine

2. Phosphatidylcholine

1

2

10214

Oils / Lipids

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UHPLC Column: ® )

Mobile Phase:

Flow Rate:

Detection:

Column Temp:

ELSD 3300:

Injection:

HPLC Column: ® )

Mobile Phase:

Flow Rate:

Detection:

Column Temp:

Injection:

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600

790.361

1492.0711220.139

826.410

1012.464

507.293

1208.6 51050.523

804.370655.384

474.329 982.501

827.416

23.221 1041.482579.315 1119.553

1312.674847.384429.090752.342 1511.078

T7

T24

T19

T13

T8

T20

T11

T18-20

T17

T4

T5T26

T9

T10

Inte

nsi

ty (

co

un

ts)

m/z

Proteins & Peptides/Reversed-Phase

TIC of Tryptic Digest of Green Fluorescent Protein (GFP): 150pmol Load on 1mm i.d.

Column: Vydac®, 250 x 1mm (Part No. 238EV51)Mobile Phase: A: Acetonitrile:0.05% Trifluoroacetic Acid (2:98) B: Acetonitrile:0.05% Trifluoroacetic Acid (90:10)Gradient: Time: 0 60 %B: 5 65Flow Rate: 50µL/minDetector: ms: ABI QSTARInjection Volume: 25µL GFP digest

Mass Spectrum of Tryptic Digest of GFP

PTH Amino Acids

Column: Vydac® Denali®, 5µm, 150 x 4.6mm (Part No. 238DE5415)Mobile Phase: A: Acetonitrile:25mM Potassium Phosphate, pH3.3 (20:80) B: Acetonitrile:25mM Potassium Phosphate, pH3.3 (80:20)

Flow Rate: 1.0mL/minDetector: UV at 254nm

1. His 2. Cys Acid 3. Arg 4. Asp 5. Ser 6. Gln 7. Thr 8. Gly 9. Cys

10. Ala 11. Tyr 12. Met 13. Pro 14. Trp 15. Phe 16. Ile 17. Leu

Gradient: Time: 0 15 %B: 0 75

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0 2 4 6 8 10 12 14 16 Min.

13

2

4

5

6

1

2

3

4

0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 Min.

10369

1. Methionine Enkephalin 2. Leucine Enkephalin 3. Angiotensin II 4. Neurotensin 5. Angiotensin I 6. Substance P

Column: Alltech® Alltim HP C18, 190Å, 5µm, 250 x 4.6mm (Part No. 87679)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.08% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 20 %B: 20 40Flow Rate: 1.0mL/minDetector: UV at 220nm

Peptide Mix on 190Å

1. Monophosphopeptide (FQpSEEQQTEDELQDK)

2. [Glu1]-Fibrinopeptide B (EGVNDNEEGFFSAR)

3. Tetraphosphopeptide (RELEELNVPGEIVEpSLpSpSpSEESITR)

4. Contaminant(s)

Phosphopeptides and Fibrinopeptide

Column: Vydac® ProZap™ C18, 2.1 x 10mmHPLC System: HPG binary pumpMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in Acetonitrile:Water (80:20)Gradient: Time: 0.0 1.0 1.1 1.3 1.4 %B: 12 60 80 80 12Flow Rate: 0.8mL/minDetector: UV at 215nm

BSA Tryptic Digest10582

Amino Acids by HILIC

Column: Alltech® Alltima HP HILIC, 150 x 4.6mmMobile Phase: Acetonitrile:10mM Ammonium Acetate with 0.2% Acetic Acid (70:30)Flow Rate: 0.75mL/minDetector: ELSD


0 1

1 34

5

67

2

2 3 4 5 6 7 8 9 10 11 Min.

10582

1. Glutamic Acid 2. Leucine 3. Serine 4. Glycine 6. Lysine 7. Arginine

Column: Vydac® 218MS C18, 5µm, 150 x 0.075mm (Part No. 218MS5.07515)Mobile Phase: A: Water with 0.1% Formic Acid:Acetonitrile (95:5) B: Water with 0.1% Formic Acid:Acetonitrile (20:80)Gradient: Time: 0 5 50 60 %B: 0 0 50 100Flow Rate: 0.25µL/min

1. (Arg-8)-Vasotocin2. (Lys-8)-Vasopressin3. Oxytocin4. Leucine Enkephalin5. Angiotensin 1

0 3

1 2

3

4

5

6 9 Min.

Column: Platinum™ 100Å 53 x 7mm Rocket™ Mobile Phase: A: 0.15% TFA in Water

B: 0.13% TFA in 95% Acetonitrile:5% WaterGradient: Time: 0 10

%B 20 50Flowrate: 3.0mL/minDetector: Alltech ELSD

8874

Column: Vydac® 208TP54, C8, 5µm, 4.6 x 250mm


Gradient: 0–30% ACN with 0.1% aqueous TFA over 60min

Separation of a Tryptic Digest on 208TP54

Reversed-Phase Column

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Peptides


a®

® 3µm,EPS C18,

1

2

3

4

0 1 2 3 4 5 Min.

4 0 600 800 1000 1200 1400 1600

m/z

0

0.4

0.8

1.2

1.6

2.0

2.4

941.0

815.9874.0

765.1

1019.2

720.2

1112.1

447.0 1223.0

680.4 1358.4483.0

644.7

719.0417 0 541 9 945.8740.5 1120.26 4 0 996.2856 7 1528.3

707 5 888.669.8 1232.4

+19 +1 +17 +16 +15 +14 + 3 +12 +11 +10 +9

Peak 1 - Bovine Cytochrome C

907.1

879.2

852 9

935.4

881.1 1037.1

966.6

1000.0807.2

828.5 968 51116.4447.0

1161.2764.1

425.11266.8893.4

1319.3784.9744.1

1209.4416.9 871.5 948.1765.91038.3

1451.1505.0955.7726.1 799 2482.9 912.0 1060.1 1160 3

1381.9978.6689.3593 2467

1323.41052.5 1131.7691.8 930.7 1527.6992.3 1353.0525.9

630.3 776.6 1 52.4590.8 1612.2418.71330.01231 8 1477.2

1363.2

+ + + 0 + + + 7 + + + + + 2 +4 + +3 +3 +3 +3 +35 +34 +3 +32 +31 +30 +29 +28 +27 +26 +25 +24 +23 +22 +21 +20 +19

Peak 3 - Bovine Carbon c Anhydrase II

400 600 800 1000 1200 1400 1600

m/z

0 2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0

1. Bovine Cytochrome C, M =12222 (ave.) 2. Horse Apomyoglobin, M = 16941 (ave.) 3. Bovine Carbonic Anhydrase II, M = 28998 (ave.) 4. Chicken Ovalbumin, M = 44455 (ave.)

Column: Vydac® ProZap™ C18, 2.1 x 10mm

Mobile Phase: A: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (5:95)

B: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (80:20)Gradient: Time: 0 4.0 4.5 4.7

%B: 20 90 90 20


Detector: AB/MDS Sciex Q TRAP® MS. EMS scan 400-1700 amu;

scan rate = 1000 amu/s; ion spray voltage = 5000 V; LIT fill time = 50 ms;

Q3 entry barrier = 8V; declustering potential = 20; collision energy = 10 eV;

curtain gas = 50; ion source gas 1 = 45; ion source gas 2 = 70;

temperature (TEM) = 350°C.

50 ng each

400 600 800 1000 1200 1400 1600

m/z

1

2

3

4

5

700 900 1100 1300 1500 1700

m/z

0

1

2

3

4

940 7

1012.2

961 8

1048.0997.7 1163.3

1140.31077 2 1348.31054.7

1482.81235.6 1521.6955.4

1387.9005.1919.3 1131.4615.9

1 34.11229.1967.5 1639.3

908.3 1117.81088.1 1396.01257.0

1200.3 1592 7945.8872.4

1282.0897.6 1351.11114.9 1515.4

1184.7 1471.9729.9 882 8 1643.9678.8621.0

1355.0 1 64.9 1527.01313.2

895.8 1480.2736.9 774.1 1248 2635.8 1655.9

1520.3 1637.8

694.5 1365.2753.0 843.5 1610.71502.3669.8

1569.6

1664.3

+35 +34 +33 +32 +31 +30 +29

Peak 4 - Chicken Ovalbumin

892.6848 1

942.2

808.0

997.2

1059.7

771.3

1304.01130.3737.9

1210.7

1412.6447.0 707.0 1322.61132.8895.2679.1 1000.9740.1425.0 805.0 1541.01216.9519.0 1694.9960.7541.0 715.6 1090.7 1245.9 1411.61011.8 1187.8 1578.6

+27 +26 +25 +24 +23 +22 +21 +20 +19 +18 +17 +16 +15 +14 +13 +12 +11

Peak 2 - Horse Apomyoglobin

LC-MS of Proteins



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Injection Amount:

1

2

3

0 1 2 3 4 5 Min.

400 600 800 1000 1200 1400 1600

0.2

0.4

0.6

0.8

1.0

1451.61161 4

968.1

1453.1

1474 5827.6

1180.41158.0446.0 651.8 969 8608.8 1262.31040.1755.6 1465.1943.6 1340 1 150 .6817.9 1685.4

+7 +6 +5

Peak 1 -Human Insulin

m/z

Inte

nsi

ty (

x 1

06 c

ps)

400 600 800 1000 1200 1400 1600

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0

1474.5

1579.71382.5

1302.1

1165.4

1229.11107.4

1054.4

1164.6

447.01006.3

1231.31108.6 1388.7

962.6483 0

504.9 664.21307.34 8.9

1201.1964.81136 3714 9

1374.0922.4524.9 1587.4885 6602.9

629.7 1493.81248.4 1396.8989.5 1127.3 1604.4

1032.9822.31528.7

+26 +25 +24 +23 +22 +21 +20 +19 +18 +17 +16 +15

Peak 3 - Human Growth Hormone

Inte

nsi

ty (

x 1

05 c

ps)

m/z

Inte

nsi

ty (

x 1

05 c

ps)

1100 1200 1300 1400 1500 1600 1700

0.4

0.8

1.2

1.6

2.0

1473.4

1542.8

1478 81387.2

1408 81446.9

1620.61331 9 1415 81513.0

1360.0 1444.91305.0

1356 5 1547.81281.2 1393.91389.7

1475.71509.8

1329.2 1420.91621.81255.1

1399.61452.21335.1

1282.7 1552 81492 8 585 01256.1

1464.81403.51363.5

1529.41323.6

1582.6 1646.61240.2 1489.31423.91213.7

1317.51537.4

1265.11599.81266.71189.2 1657 51505 8

1319.31376.3 1576.4

1168.5 1270.71694.31642.91220.2

1130.41150 7 1567.8

1143.7108 .51016 5 1678 7

+59 +58 +57 +56 +55 +54 +53 +52 +51 +50 +49 +48 +47 +46 +45 +44 +43 +42 +41

Peak 2 - Human Serum Albumin

m/z

1. Human Insulin, M = 5803 (ave.) 2. Human Serum Albumin, M = 66520 (ave.) 3. Human Growth Hormone, M = 22123 (ave.)

LC-MS of Proteins

Column: Vydac.® ProZap™ C18, 2.1 x 10mmMobile Phase: A: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (5:95) B: 0.2% Formic Acid, 0.01% Trifluoroacetic Acid in Acetonitrile:Water (80:20)Gradient: Time: 0 4.0 4.5 4.7 %B: 20 90 90 20 Flow Rate: 0.4mL/minDetector: AB/MDS Sciex Q TRAP® MS. EMS scan 400-1700 amu; scan rate = 1000 amu/s; ion spray voltage = 5000 V; LIT fill time = 50 ms; Q3 entry barrier = 8V; declustering potential = 20; collision energy = 10 eV; curtain gas = 50; ion source gas 1 = 45; ion source gas 2 = 70; temperature (TEM) = 350°C.Injection Amount: 8µg each, except 1.5µg for growth hormone



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20 40 60Min.

VYDAC® MS C4 Column1

2

Competitor 1 C4 Column1

2


2

Competitor 3 C3 Column1 2

VYDAC® MS C4 Column

1

2

3

4


3

4


2

3

4


42

Competitor 4 C4 Column1 2

4

20 65 Min.30 40 50 60

20 40 60 80 Min.

VYDAC® MS C18

Column

Competitor 1 C18

Column

Competitor 2 C18

Column

Columns: Vydac® MS C4 (Part No. 214MS54), Competitor 1 C4, Competitor 2 C4, and Competitor 3 C3 (all 300Å, 250 x 4.6mm packed with 5µm except 4µm for Competitor 1)Mobile Phase: 50mM Tris, pH7.5:n-Propanol (71:29)Flow Rate: 0.5mL/minDetector: UV at 220nm

Human Growth Hormone (hGH) Assay on 300Å C4 Columns

Columns: Vydac® MS C4 (Part No. 214MS54), Competitor 1 C4, Competitor 2 C3, Competitor 3 C4, and Competitor 4 C4 (all 300Å, 250 x 4.6mm packed with 5µm except 4µm for Competitor 1)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient Vydac®, Comp. 1, & Comp. 2: Time: 0 20 25 45 %B: 20 60 80 80Gradient Comp. 3 & Comp. 4: Time: 0 20 25 45 50 80 %B: 20 60 80 80 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm

Transmembrane Protein p14 on 300Å C4 Columns 1. 18.8 kD Protein 2. Non-myristoylated p14 3. Myristoylated p14 4. Triton™ X (Surfactant)

1. Desamido hGH 2. hGH

Columns: Vydac® MS C18 (Part No. 238MS5415), Competitor 1 C18, and Competitor 2 C18 (all 300Å, 5µm, 150 x 4.6mm)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 5 80 90 100 %B: 4 4 40 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm

Tryptic Digest of Fetuin on 300Å C18 Columns



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0

1

2

3

4

5

6

7

0 5 15 25 35 45 55 65 75 85 95 Min.

Inte

nsi

ty (

x 1

06 c

ps)

1. Chicken 2. Bovine 3. Ovine 4. Rabbit 5. Human 6. Porcine 7. Rat I 8. Rat II

Base Peak Chromatogram of

Tryptic Digest of BSA

Insulin Variants

1. ApoC-IIIa 2. ApoC-IIIb 3. ApoC-IIa 4. ApoC-IIIc 5. ApoC-I 6. ApoC-IIb 7. ApoA-Ia 8. ApoA-Ib 9. ApoA-IIc 10. ApoA-IIa 11. ApoA-IIb

Human Apolipoproteins

Column: Vydac® Everest® C18, 300Å, 5µm, 300µm x 150mm (Part No. 238EV5.315)Mobile Phase: A: 0.1% Formic Acid in Water B: 0.1% Formic Acid in Acetonitrile Gradient: Time: 0 5 75 85 95 %B: 4 4 40 90 90Flow Rate: 5µL/minColumn Temp: 22ºCPeptide Load: 0.1µL injection of 5.8µg/µL (580ng or 9pmol)MS Detector: AB/MDS Sciex Q-Trap.

Tryptic Digest of Bovine Serum Albumin Tryptic Digest of Fetuin (a Glycoprotein)

Column: Vydac® C18, 5µm, 250 x 4.6mm (Part No. 218TP54)Mobile Phase: A: Acetonitrile:0.1% Trifluoroacetic Acid (25:75) B: Acetonitrile:0.1% Trifluoroacetic Acid (58:42)Gradient: Time: 0 33 %B: 0 100Flow Rate: 1.2mL/minDetector: UV at 215nm

Column: Vydac® C4, 5µm, 250 x 4.6mm (Part No. 214TP54)Mobile Phase: A: 0.1% Trifluoroacetic Acid B: IsopropanolGradient: Time: 0 355 %B: 10 38Detector: UV at 220nm

Column: Vydac® Everest® C18, 5µm, 150 x 4.6mm (Part No. 238EV5415) Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 5 80 90 100 %B: 4 4 40 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm

Column: Vydac® Everest® C18, 5µm, 150 x 4.6mm (Part No. 238EV5415)Mobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 5 80 90 100 %B: 4 4 40 90 90Flow Rate: 1.0mL/minDetector: UV at 215nm

Column: Vydac® C4, 5µm, 250 x 4.6mm (Part No. 214TP54)Mobile Phase: A: 0.1% Trifluoroacetic Acid B: AcetonitrileGradient: Time: 0 25 %B: 27 30Detector: UV at 210nm

Ribosomal Proteins



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Peptides on 238MS C18 Column with Various Modifiers

In order to demonstrate the effects of mobile phase modifiers and modifier concentrations, a mixture of six peptides was separated on a monomeric C18 MS column under identical gradient conditions with a variety of ion-pair reagents. Significant differences in selectivity arise depending on the concentrations and identities of the ions present in the mobile phase. Perfluorinated Acidic modifiers even at very low levels (0.005% w/v) produce increased retention. For this particular mixture, the best spread of peaks was obtained with heptafluorobutyric Acid (HFBA) at 0.02% (w/v). However, this might be expected to differ for other peptides.

Column: Vydac® Monomeric C18, 5µm, 250 x 4.6mm (Part No. 238MS54)Gradient: 10 to 40% Acetonitrile over 30 min for all chromatograms with constant concentration of modifier as indicated (w/v) Flow Rate: 1.0mL/minDetector: UV at 220nmSample: Oxytocin, Bradykinin, Angiotensin II, Eledoisin- related Peptide, Neurotensin, and Angiotensin I

0 0.2 0.4 0.6 0.8 1.0 1.2 Min.

5

2

1

24

3

2 4 53

1

0 0.2 0.4 0.6 0.8 1.0 Min.

1

2

0 1 2 3 4 5 6 Min.

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 Min.

1

2

3

4

5

6

7

Column: Vydac® ProZap™ C18, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid in AcetonitrileGradient: Time: 0 4.0 4.5 4.7 %B: 23 75 75 23 Flow Rate: 0.2mL/minDetector: UV at 280nm

Column: Vydac® ProZap™ C18, 500Å, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid:Acetonitrile (20:80)Gradient: Time: 0 0.1 0.7 1.1 1.2 %B: 4 15 20 50 4Flow Rate: 0.8mL/minDetector: UV at 215nm

1. Albumin 2. IgG

Antibodies in Sheep Serum on ProZap

Column: Vydac® ProZap™ C18, 500Å, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid:n-Propanol (10:90)Gradient: Time: 0 6.0 6.5 7.0 %B: 5 75 75 5Flow Rate: 0.5mL/minDetector: UV at 280nm

1. GY (238 Da) 2. VYV (379 Da) 3. Met Enkephalin (YGGFM, 573 Da) 4. Leu Enkephalin (YGGFL, 555 Da) 5. Angiotensin II (DRVYIHPF, 1045 Da)

Peptides

Column: Vydac® ProZap™ C18, 500Å, 1.5µm, 10 x 2.1mmMobile Phase: A: 0.1% Trifluoroacetic Acid in Water B: 0.085% Trifluoroacetic Acid:Acetonitrile (20:80)Gradient: Time: 0 0.1 0.7 1.1 1.2 %B: 4 15 20 50 4Flow Rate: 0.8mL/minDetector: UV at 215nm

1. Contaminant Peptides/Impurities 2. RGAGGLGLGK-amide (883 Da) 3. Ac-RGAGGLGLGK-amide (926 Da) 4. Ac-RGVGGLGLGK-amide (954 Da) 5. Ac-RGVVGLGLGK-amide (996 Da)

Synthetic Peptides-10 Injections

1. Ribonuclease (37ng) 2. Bovine Insulin (37ng) 3. Cytochrome C (37ng) 4. Lysozyme (4ng) 5. Bovine Serum Albumin (37ng) 6. Carbonic Anhydrase (37ng) 7. Ovalbumin (37ng)

Fast Protein Analysis


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Column: Vydac® C18, C4 and Diphenyl, all 5µm, 150 x 4.6mm

(Part Nos. 218TP5415, 214TP5415, 219TP5415)

Mobile Phase: A: 0.1% Trifluoroacetic Acid

B: Acetonitrile


B%: 10 90

Sample: 18-Residue Helical Peptide (1) and

Six-helix Template- assembled Synthetic Protein (2)



Purification of Trityl-On Oligonucleotides

Comparison of Polypeptide Separations on

C18, C4 and Diphenyl Reversed-Phase Columns

1. H 2. TH6

Column: Vydac® Diphenyl, 5µm, 250 x 4.6mm

(Part No. 219TP54)


B: n-Propanol:Acetonitrile:0.1% Trifluoroacetic Acid (50:50)


%B: 25 70



Separation of Protein / Peptide Mixture on

Vydac® 219TP Diphenyl Column

1. Horse Cytochrome C Tryptic Digest Fragments 2. Whale Myoglobin CNBr Digest Fragments 3. Ribonuclease 4. Cytochrome 5. Lysozyme 6. BSA 7. Myoglobin 8. Ovalbumin

Column: Vydac® C4, 5µm, 250 x 10mm (Part No. 214TP510)

Mobile Phase: A: 0.1M Triethylammonium Acetate, pH7.0 B: Acetonitrile


B%: 16 23



Analysis of a Lipid Peptide

Column: Vydac® Diphenyl, 5µm, 250 x 4.6mm

(Part No. 219TP54)


B: Acetonitrile

Gradient: Time: 0 67.5

B%: 0 70




Columns: Vydac® 218TP54 and 238TP54, both C18, 5µm,

4.6 x 250mm


Gradient: 10–40% ACN with 0.1% TFA (w/v) over 30min


Synthetic Peptides 1. GY 2. VYV 3. Neurotensin fragment 1-8 (pELYENKPR) 4. Ac-RGGGGLGLGK-NH2 5. RGAGGLGLGK-NH2 6. Ac-RGAGGLGLGK-NH2 7. Ac-RGVGGLGLGK-NH2 8. Oxytocin (CYIQNCPLG-NH2) 9. Met Enkephalin (YGGFM) 10. Bradykinin (RPPGFSPFR) 11. Ac-RGVVGLGLGK-NH2 12. Neurotensin Fragment 8-13 (RRPYIL) 13. Angiotensin II (DRVYIHPF) 14. Leu Enkephalin (YGGFL) 15. Neurotensin (pELYENKPRRPYIL) 16. Angiotensin I (DRVYIHPFHL)

Human Growth Hormone

1. di-Desamido HGH2. mono-Desamido HGH3. Human Growth Hormone (HGH)


Mobile Phase: Isopropanol:0.01M Tris buffer, pH7.5 (29:71)





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Min.

Analyses

Startingmaterial

Pure pool(fract 2 -16)

Impure pool(fract 1 &17-26)

Fraction Collectionfrom Run C

Fraction 1

Fractions 2-16

Fractions 17-26

A280

A) 26mg

B) 41.5mg

C) 60mg

0 25 7550 100 Min.

0 5 10 15 Min.

6x scale

6x scale

6x scale

40 60 807050 90 Min.

Column: Vydac® C4, 10-15µm, 10 x 250mm (Part No. 214TP101510)

A) Sample: 26mg partially purified porcine insulin in 30mL of 20%

Acetonitrile/0.1% Trifluoroacetic acid, pumped on at 8mL/min

Elution: 4mL/min

Gradient: 20-35% Acetonitrile in 0.1% Trifluoroacetic acid over 90 min

B) Sample: 41.5mg in 42mL of 20% Acetonitrile/0.1% Trifluoroacetic

acid, pumped on at 8mL/min

Elution: 4mL/min

Gradient: 20-27% Acetonitrile in 35 min, hold 27% for 10 min

followed by 27-28% in 20 min, 28%-35% in 10 min, and

35-95% in 5 min, all in 0.1% Trifluoroacetic acid.

C) Sample: 60mg in 60mL of 20% Acetonitrile/0.1% Trifluoroacetic

acid, pumped on at 8mL/min

Elution: 4mL/min

Gradient: 20-27% Acetonitrile in 35 min, hold 27% for 10 min

followed by 27-28.5% in 30 min, hold 28.5% for 20 min

Preparative Chromatography of Porcine Insulin Analyses of Starting Material and Pooled

Fractions from Preparative Column

Column: Vydac® C4, 5µm, 4.6 x 150mm

(Part No. 214TP5415)

Samples: Starting material and pooled fractions

of preparative run C, as indicated.

Detection: UV at 215nm

Elution: 1.5mL/min

Gradient: 30-33% Acetonitrile in 0.1%

Trifluoroacetic acid over 15 min


0 1 2 Min.

1

2

3

4

5

6

7

Expedite™ Columns for Low Volume High Throughput LC

Column: ProZap™ C18, 1.5µm, 2.1 x 10mm Expedite™

Mobile Phase: A: 0.1% TFA in Water

B: 0.085% TFA in Acetonitrile



Injection Vol.: 5µL

1. Ribonuclease (37ng) 2. Bovine Insulin (37ng) 3. Cytochrome C (37ng) 4. Lysozyme (4ng) 5. Bovine Serum Albumin (37ng) 6. Carbonic Anhydrase (37ng) 7. Ovalbumin (37ng)

10736


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Column: Astec™ Chirobiotic® T™, 250 x 4.6mm (Part No. 12053)

Mobile Phase: Ethanol:Water (80:20)



Phenylalanine Enantiomers


Mobile Phase: A: 5mM Heptafluorobutyric Acid, 0.7% Trifluoroacetic Acid

B: Acetonitrile


%B: 0 0 15 35


Detector: ELSD


(Part No. 32214)




10. Arg 11. Val 12. Met 13. Tyr 14. Ile 15. Leu 16. Phe 17. Trp

1. Gly 2. Ser/Asn 3. Asp 4. Gln 5. Ala/Thr 6. Glu 7. Cys/Lys 8. His 9. Pro

Amino Acids

Amino Acids without Derivatization

Thyroid Chemicals

8770

8282

1. DL-Tyrosine 2. 3-Iodo-Tyrosine 3. 3,5- Diiodo-D-Tyrosine

4. DL-Thyroxine 5. 3,3,5-Triiodo-L-Thyronine

Column: Standard Platinum™ 100Å C18, 3µm,53 x 7mm Rocket™ Column

Mobile Phase: A: 0.15% TFAB: Acetonitrile

Gradient: Time: 0 3 12 15%B 3 3 65 65

Flowrate: 2.5mL/minDetector: Alltech ELSD

Drift Tube Temp: 40°CNitrogen Flowrate: 1.75SLPM

1. Noradrenaline (norepinephrine)2. Adrenaline (epinephrine)3. Dopamine (3-Hydroxytyramine)4. 3,4-Dihydroxyphenylalanine (DOPA)5. Phenylalanine6. Homovanillic Acid (HVA)

0 1

1

2

2

3

3

4

4

5

5

6

6

7 Min.

8903

Catecholamines



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Amino Acids

0

1

2

3

4

56

7

8

9

10

11

13

14

1512

5 10 15 20 25 30 Min.

1. Aspartic Acid 2. Glutamic Acid 3. Serine 4. Histidine 5. Glycine 6. Threonine 7. Arginine 8. Alanine 9. Tyrosine 10. Methionine 11. Valine 12. Phenylalanine 13. Isoleucine 14. Leucine 15. Lysine

Column: OPA-HR, 5µm, 4.6 x 150mm (Part No. 28072)

Mobile Phase: A: Methanol B: Tetrahydrofuran

C: 20mM Sodium Acetate buffer, pH 5.5


%A: 5 65

%B: 5 2

%C: 90 23


Detector: Fluorescence

OPA-Derivatized Amino Acids9234

Prevail™ Bonded Phases are Stable in 100% Aqueous Conditions

Catecholamines

1. Norepinephrine2. Epinephrine3. 5-Hydroxydopamine

Mobile Phase: 25mM KH2PO4, pH



9450

9452

After 24 hrs.

After 8 hrs.

After 20 min.

9451

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0 5 10 15 20 Min.

3

45

1

2

0 2 4 6 8 10 12 Min.

1

23

4

5

0 2 4 6 8 Min.

1

2

3

4

5

0 2 4 6 8 Min.

1

23

4

5

10347

1. Cytidine 2. Uridine 3. Xanthosine 4. Guanosine 5. Adenosine


(Part No. 32246)



%B: 5 5 30




Column: Alltech® Alltima HP EPS C18, 5µm, 150 x 4.6mm

(Part No. 87715)

Mobile Phase: A: 0.03M Potassium Phosphate, pH3.2

B: Acetonitrile


%B: 5 5 30



8245

Nucleic Acids

NucleosidesNucleosides



Flow Rate: 1mL/min

Detector: UV at 254





Detector: UV at 254

1. Cytosine 2. Uracil 3. Guanine 4. Adenine 5. Thymine

94119410

Nucleosides Nucleic Acid Bases



scie

nce

® ®

1

23

4

5

6

7

8

9

10

11

12

0 4 8 12 16 20 24 28 Min.

0 3

1

2

3

4

5 6

6 9 12 15 18 21 24 Min.

Column: Alltech® Platinum™ EPS C18, 3µm, 53 x 7mm Rocket™

(Part No. 50573)




1. Uracil 2. Benzamide 3. Benzyl Alcohol 4. Phenol 5. Methylbenzoate 6. Acetophenone 7. N,N-Diethyl-m-toluamide 8. Toluene 9. Benzophenone 10. Biphenyl

8982

Monoaromatics

7373

54409538





Column: Alltech® OA-1000, 9µm, 300 x 6.5mm (Part No. 9046)


Flow Rate: 0.8ml/min

Column Temp: 65°C

Detector: RI

1. Glycerol 2. Methanol 3. Ethanol 4. Isopropanol 5. tert-Butanol 6. Propanol 7. sec-Butanol 8. Isobutanol 9. tert-Pentanol 10. Butanol 11. Isopentanol 12. Pentanol

1. 3-Amino-4-hydroxybenzoic Acid 2. 4-Hydroxybenzoic Acid 3. Benzoic Acid 4. 4-Phenylbutyric Acid 5. 5-Phenylbutyric Acid


(Part No. 88640)




10314

1. 1,2-Phosphatidyl-ethanolamine 2. Phosphatidyl-DL-glycerol 3. Phosphatidylinositol 4. 1,2-Phosphatidyl-L-serine 5. Phosphatidylcholine 6. Sphingomyelin

1. DL-4-Hydroxy-3-methoxy-mandelic Acid 2. Homovanillic Acid 3. Vanillic Acid 4. Salicylic Acid 5. Benzoic Acid


Mobile Phase: A: Isopropanol B: Hexane C: Water


%B: 40 40 40

%C: 2 8 8


Detector: ELSD

Phospholipids

Miscellaneous

Benzyl Acids

Aromatic Acids

Alcohols



scie

nce

®

0 1

1

2

3

4 5

6

7

2 3 4 5 Min

0 3

1

2

3

4

6 9 12 Min

0 2 4 6 8 10 12 14 16 18 20 Min.

2

13

45

6

7

10346

Column: Alltech® Platinum™ EPS C18, 1.5µm, 33 x 7mm Rocket™ (Part No. 50575)Mobile Phase: 0.025M KH2PO4, pH3.0:Acetonitrile (65:35)Flow Rate: 2.0mL/minDetector: UV at 254nm

Column: Alltech® Alltima HP EPS C18, 5µm, 150 x 4.6mm (Part No. 87715)Mobile Phase: Water:Methanol (60:40)Flow Rate: 1.0mL/minDetector: UV at 214nm

8671

Column: Alltech® Platinum™ C18, 5µm, 150 x 4.6mm (Part No. 32043)Mobile Phase: 0.025M K2HPO4, pH7.0:Methanol (25:75)Flow Rate: 1.0mL/minDetector: UV at 220nm

1. Methoxychlor 2. Dieldrin 3. Heptachlor 4. 4,4-DDT

8437

1. Simazine 2. Simetryn 3. Prometon 4. Atrazine 5. Ametryn 6. Prometryn 7. Terbutryn

8439

1. Diquat 2. Paraquat

Column: Alltech® Platinum™ Silica 100Å 3µm, 30mm x 4.6mm (Custom)Mobile Phase: 0.01M Tetramethylammonium Hydroxide and 0.15M Ammonium Sulfate in Water, pH3.0 with 5M Sulfuric AcidFlow Rate: 0.5mL/minDetector: UV at 310nm until 3 minutes UV at 260nm after 3 minutes

10355

Column: Alltech® Alltima HP CN, 5µm, 150 x 4.6mm (Part No. 87769)Mobile Phase: Water:Acetonitrile (60:40)Flow Rate: 1.0mL/minDetector: UV at 254nm

1. Fenuron 2. Monuron 3. Diuron 4. Linuron

1. Methyl Parathion 2. Dichlorvos 3. Ethoprophos 4. Azinphos

10739

1. Fenuron 2. Monuron 3. Fluometuron 4. Diuron 5. Methiocarb 6. Linuron 7. Neburon

Column: GraceSmart™ C18, 5µm, 150 x 4.6mm (Part No. 5138812)Mobile Phase: A: 0.025M KH2PO4, pH3.2 B: AcetonitrileGradient: Time: 0 20 %B: 30 60Flow Rate: 1.0mL/minDetector: UV at 240nm

Pesticides

Organophosphates Organochlorines

Triazine Herbicides Herbicides

Bipyridylium HerbicidesCarbamate and Urea Pesticides


hplc applicationsenvironmental

®

®

n n

n n

0 2 4 6 8 10 12 14 Min.

1

2

0 1 2 3 4 5 6 7 8 9 Min.

1

2

34

56

7 89

10740

1. Methoxychlor 2. 4,4-DDT


(Part No. 5138812)

Mobile Phase: Methanol:0.025M K2HPO4, pH7.0 (75:25)



Organochlorine Pesticides

10580

Column: Vydac® Denali® 238DE C18, 3µm, 100 x 4.6mm

(Part No. 238DE3410)

Mobile Phase: A: 25mM Potassium Phosphate, pH6.4

B: Acetonitrile

Gradient: Time: 0 0.5 7.5 8.0

%B: 40 40 63 63



1. Simazine 2. Atraton 3. Simetryn 4. Atrazine 5. Prometon 6. Ametryn 7. Propazine 8. Prometryn 9. Terbutryn

Triazine Pesticides

Column: Vydac® Denali®, 3µm, 100 x 4mm (Part No. 238DE3410)

Mobile Phase: A: 0.5% Phosphoric Acid Buffer, pH1.8; B: Acetonitrile

Gradient: Time: 0 5

%B: 50 90



Separation of Chlorophenoxy Acid Herbicide Mixtures10713

Benzenes

1. Thiourea 2. Toluene 3. Propylbenzene 4. Butylbenzene


(Part No. 5139605)

Mobile Phase: 20mM Potassium Phosphate, pH2.5:Acetonitrile (45:55)



Benzenes

1. Picloram 2. Chloramben 3. Dicamba 4. Bentazon 5. 2,4-D 6. Dichlorprop 7. 2,4,5-TP 8. Acifluorphen

0 0.5 1 1.5 2 Min.


enviro

nmental

hplc applications

®

1

2

3

4

5 Min.

12 3

4

5

6

78

910

11

12

13

1415

16

0 1 2 3 4 5 6 Min.




%B: 72 72 100


Detector: UV at 254

1. Benzene 2. Naphthalene 3. Acenaphthylene 4. Fluorene 5. Acenaphthene 6. Phenanthrene 7. Anthracene 8. Fluoranthene 9. Pyrene

10. Benz[a]anthracene 11. Chrysene 12. Benzo[b]fluoranthene 13. Benzo[k]fluoranthene 14. Benzo[a]pyrene 15. Dibenz[a,h]anthracene 16. Benzo[g,h,i]perylene 17. Indeno[1,2,3,c,d]pyrene

Column: Alltech® Alltima HP C18 HiLoad, 3µm, 53 x 7mm

(Part No. 87690)


Gradient: Time: 0 0.1 5

%B: 40 40 100

Flow Rate: 5mL/min


9400

1. Naphthalene 2. Acenapthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene

7. Fluoranthene 8. Pyrene 9. Benzo[a]anthracene 10. Chrysene 11. Benzo[b]fluoranthene 12. Benzo[k]fluoranthene

10757

PAHsPolyaromatic Hydrocarbons

Fast Polyaromatic Hydrocarbons

1. Naphthalene 2. Acenaphthylene 3. 1-Methylnaphthalene 4. 2-Methylnaphthalene 5. Acenaphthene 6. Fluorene 7. Phenanthrene 8. Anthracene

9. Fluoranthene 10. Pyrene 11. Benzo[c]phenanthrene 12. Cyclopenta[c,d]pyrene 13. Benz[a]anthracene 14. Chrysene 15. Benzo[b]naptho[2,1-d]thiopen 16. 7,12-Dimethylbenz[a]anthracene

Beyond the EPA 16 Priority-Pollutant PAHs




%B: 50 100


13. Benzo[a]pyrene 14. Dibenzo[a,h]anthracene 15. Benzo[g,h,i]perylene 16. Indeno[1,2,3,c,d]pyrene

17. Benzo[e]pyrene 18. Benzo[b]fluoranthene 19. Benzo[k]fluoranthene 20. Benz[a]pyrene 21. Dibenz[a,h]anthracene 22. Benzo[g,h,i]perylene 23. Indeno[1,2,3,c,d]pyrene

Detector: UV at 254


hplc applicationsenvironmental

®

PAHs

10358

Column: Alltech® Alltima HP CN, 5µm, 150 x 4.6mm

(Part No. 87769)

Mobile Phase: Hexane:Isopropanol (85:15)



1. Nitrobenzene 2. 2,6-Dinitrotoluene 3. 1,3-Dinitrobenzene 4. 4-Nitrophenol

Nitroaromatics

Nitroaromatics (Normal-Phase)

1. Naphthalene 2. Acenaphthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene

Column: Vydac® C18, 5µm, 4.6 x 150mm

(Part No. 201TP5415)


Mobile Phase: A: Water B: ACN

Gradient: Hold 50% B for 3min, then 50 to 100% B in 7min


Priority Pollutants PAHs

In Accordance with

EPA Methods 505, 550.1, 610, and 8310

9. Benz[a]anthracene 10. Chrysene 11. Benzo[b]fluoranthene 12. Benzo[k]fluoranthene 13. Benzo[a]pyrene 14. Dibenz[ah]anthracene 15. Benzo[ghi]perylene 16. Indeno[1,2,3-cd]pyrene

enviro

nmental


hplc applications

®

0 5 10 15 20 25 30 35 40 Min.

12

34

5

6

7

89

10

11

12 13 14

0

1

3

4

5

67

8

9

10

111213

15

16

14

17

18

192

10 20 30 40 50 Min.

2

1

34

5

0 2 4 6 8 10 12 14 Min.

0 2 4 6 8 10 12 14 16 18 20 22 Min.

12

3

4

5

6

7

89

10 11 12

13


(Part No. 32246)

Mobile Phase: A: 0.03M Monosodium Phosphate, pH2.5

B: Acetonitrile:Methanol:Water (40:50:10)

Gradient: Time: 0 3 13 20 25 30 40 45 55

%B: 5 5 10 10 20 40 75 80 80



1. 4-Amino-5-hydroxy-2,7-naphthalene-disulfonic Acid 2. Hydroquinone 3. Benzylamine 4. 3'-Aminoacetophenone 5. Phenol 6. 4-Hydroxyphenylacetic Acid 7. 1-Naphthalene Sulfonic Acid 8. 2,4-Dihydroxybenzoic Acid 9. 1-Naphthalene Sulfonic Acid Degradant 10. 4-Hydroxy-3-methoxybenzoic Acid 11. Phenylacetic Acid 12. m-Cresol 13. 4-Hydroxycinnamic Acid 14. trans-Cinnamaldehyde 15. trans-Cinnamaldehyde Degradant 16. trans-Cinnamaldehyde Degradant 17. 2-Naphthalenethiol Degradant 18. 2-Naphthalenethiol 19. 2-Naphthalenethiol Degradant

Column: Alltech® Adsorbosphere™ UHS C18, 5µm, 150 x 4.6mm

(Part No. 288118)




1. HMX

2. RDX

3. 1,3,5-TNB


(Part No. 5138810)

Mobile Phase: A: 1% Glacial Acetic Acid in Water

B: 1% Glacial Acetic Acid in Methanol


%B: 45 45 60 80



4. 1,3-DNB 5. Tetryl 6. Nitrobenzene (NB) 7. 2,4,6-TNT

10511

8447

10707

Phenols

1. Phenol

2. 4-Nitrophenol

3. 2-Chlorophenol

4. 2-Nitrophenol

5. 2,4-Dichlorophenol

PhenolsSeparation of EPA 604 Phenols Mixture

1. Phenol 2. 4-Nitrophenol 3. 2-Chlorophenol 4. 2-Nitrophenol 5. 2,4-Dinitrophenol 6. 2,4-Dimethylphenol 7. 4-Chloro-3-Methylphenol 8. 2,4-Dichlorophenol 9. 2-Methyl-4,6-Dinitrophenol 10. 2,4,6-Trichlorophenol 11. Pentachlorophenol


Mobile Phase: A: 1% Acetic Acid in Water

B: 1% Acetic Acid in Acetonitrile


%B: 35 63 95 95




(Part No. 32064)




1. HMX

2. RDX

3. TNB

4. DNB

5. Tetryl

6. NB

7. TNT 8. 4-A-DNT 9. 2,6DNT 10. 2,4DNT 11. 2-NT 12. 3-NT 13. 4-NT

8279

8. 4-Amino-2,6-Dinitrotoluene (4-AM-DNT) 9. 2-Amino-4,6-Dinitrotoluene (2-Am-DNT) 10. 2,6-DNT 11. 2,4-DNT 12. 2-NT 13. 4-NT 14. 3-NT

Water PollutantsSemipolar & Highly Hydrophilic Waste Water Contaminants

Explosive Analysis, EPA Method 8330

Explosives

Nitroaromatic and Nitroamine Explosives

environmental

57

hplc applications


Water Pollutants

8

Fullerenes

Column: Vydac® C4, 5µm, 250 x 10mm (Part No. 201TP510)

Mobile Phase: Acetonitrile:Toluene (50:50)

Flow Rate: 5mL/min

Buckminsterfullerenes

10310 1. n-octyl Glucopyranoside (50ng on column) 2. n-decyl Glucopyranoside (50ng on column) 3. n-dodecyl Glucopyranoside (50ng on column)




Detector: ELSD

Microbore Analysis of Glucopyranosides

Surfactants

0 1

1

2

2

3

3

4 5 6 7 8 Min.


(Part No. 88056)



Detector: ELSD

8636

Polymers

0 4 8 12 16 20 Min.

Dimethicone

0 5 10 15 20 25 30 Min.

7809

Tween 80

Column: Alltech® Adsorbosphere™ C8, 5µm, 250 x 4.6mm

(Part No. 287112)

Mobile Phase: A: Methanol B: Water


%B: 60 20


Detector: ELSD

8442

Polyethylene Glycol (MW=600)

0 5 10 15 20 25 30 35 Min.

miscella

neous


hplc applications

®


Mobile Phase: A: Acetonitrile B: Chloroform


%B: 25 75


Detector: ELSD

®®

Column: Alltech® Alltima C18, 3µm, 20 x 2.1mm Expedite™

(Part No. 43803)



Gradient: Time: 0 2

%B: 5 100



Columns: Alltech® Alltima C18, 3µm, 20 x 4.6mm Expedite™

(Part No. 43851)



Gradient: Time: 0 2

%B: 5 100



9462

9467

1. Uracil2. Theophylline3. Acetophenone4. Propiophenone5. Butyrophenone6. Benzophenone7. Valerophenone8. Heptanophenone9. Octanophenone

1. Uracil2. Theophylline3. Acetophenone4. Propiophenone5. Butyrophenone6. Benzophenone7. Valerophenone8. Heptanophenone9. Octanophenone

Phenones/Fast Gradient Column Formats Phenone Homologs

Phenone Homologs

1

0 1 2 3 Min.

2 3

4 5

6

7

89

0 1 2 3 Min.

1

2

3

4 5

6

7

8

9

Column: Alltech® Prevail™ C18, 3µm, 20 x 2.1mm, Expedite™

(Part No. 43827)




%B: 0 100 100



9460

1. Uracil 2. Theophylline 3. Acetophenone 4. Propiophenone 5. Butyrophenone 6. Benzophenone 7. Valerophenone 8. Heptanophenone 9. Octanophenone

Phenone Homologs

miscellaneous

59

hplc applications


® ®

0 105 25 Min.2015

12

3

5

4

6

9

10

1213

11

78

0 105 25 Min.2015

1

2

3

5

6

7

8

9

10

11 12 1314

15

4

0 5 10 15 20 25 30 Min.

1

9

8

76

5

4

32

1011

0 105 25 Min.2015

1 15

14

13

12

11

10 9

7

8

6

3

4

5

2

0 105 25 Min.2015

1

2

34

5

6

78

910

Biofuels

10644


(Part No. 87698)



%B: 0 15 70 70 0


Detector: ELSD


(Part No. 87698)



%B: 30 70 70 30 30


Detector: ELSD

10748

1. LLnLn 2. LLLn 3. LLL 4. OLLn 5. PLLn 6. LLO 7. PLL 8. LLO

9. LLS 10. PLO 11. PLP 12. OOO 13. SLO 14. POO 15. POP

10749


(Part No. 87698)



%B: 30 70 70 30 30


Detector: ELSD


(Part No. 87698)



%B: 30 70 70 30 30


Detector: ELSD

Column: Alltech® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm (Part No. 87698)



%B: 30 70 70 30 30


Detector: ELSD

10750

1. PLL 2. LOO 3. PLO 4. PLP 5. MOP 6. OOO 7. POO 8. POP 9. SOO 10. PPP 11. POS 12. PPS 13. SOS

10751

1. LLnLn 2. LLLn 3. OLnL 4. LLO 5. LOO 6. PLO 7. OOO 8. POO 9. POP 10. PPP

1. LLnLn 2. LLLn 3. LLL 4. OLLn 5. PLLn 6. LLO 7. PLL 8. LOO

9. LLS 10. PLO 11. PLP 12. OOO 13. SLO 14. POO 15. POP

1. Mono-acylglycerol (MAG) 2. MAG 3. MAG

4. Fatty Acid Methyl Ester (FAME) 5. MAG 6. FAME 7. FAME 8. MAG 9. FAME 10. Di-acylglycerol (DAG) 11. Tri-acylglycerol (TAG)

Biodiesel in Spec with EN14105 Method

Triacylglycerols in Soy Oil

Triacylglycerols in Canola Oil Triacylglycerols in Red Palm Oil

Triacylglycerols in Used Chinese Frying Oil

miscella

neous


hplc applications

® ®

®

® ®

0 105 25 Min.2015

1

2

3

4

0 2 4 6 8 Min.

1

5

7

64

3

2

0 2 4 6 8 Min.

2

4

3

1

1. Hexabromo Cyclododecane 2. Tetrabromo Bisphenol A (Allyl Ether)




Detector: ELSD

10261

Fire Retardants

Brominated Flame Retardants

10752

Column: ® Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm

(Part No. 87698)



%B: 30 70 70 30 30


Detector: ELSD

1. PPO 2. PPP 3. POS 4. SSS

Triacylglycerols in Lard

Biofuels

8760

1. Nicotine 2. Quinine 3. Diphenhydramine 4. Nortriptyline 5. 2,4, Dichlorophenoxy Acetic Acid 6. 2,4, Dichlorophenoxy Propionic Acid 7. 2,4, Dichlorophenoxy Butyric Acid




10695

1. Uracil 2. Amitriptyline 3. Phenylvaleric Acid 4. Toluene




StandardsStandards Mix Acids, Bases and Neutrals

Column:

Mobile Phase: 50mM KH2PO4, pH3.0:CH3CN (40:60)

Flowrate: 1.0mL/min


1. Uracil

2. Pyridine

3. Phenol

4. N,N-Dimethylaniline

5. 4-Butylbenzoic Acid

6. Toluene

Acids, Bases, and Neutrals

1

2

3

4

6

5

0 1 2 3 4 5 Min.

1. Phenylephrine

2. Diphenhydramine

3. Amitriptyline

0 1

1

2

2

3

3

4 5 Min.

Column: VisionHT C18-B, 1.5µm, 2.0 x 50mm (Part No. 5141908)

Mobile Phase: 50mM Ammonium Formate pH 7:Methanol (20:80)

Column Temp: 40°C



miscellaneous

61

hplc applications


® ®

®

Alltech

® Alltima C18, 3µm, 100 x 4.6mm®Alltech

®

Separate Highly Basic Components at

Neutral pH with VisionHT C18-B

Standards

Column:




Final Acid washed with 24,000 column volumes of Acetonitrile:Dilute Sulfuric Acid, pH 1.0 (50:50) at 60°C.

Final Base washed with 24,000 column volumes of Acetonitrile Ammonium Hydroxide, pH 10 (50:50) at 20°C.

Retention Times

1. Uracil 0.56 0.55 0.55

2. Phenol 0.82 0.81 0.81

3. N,N-Diethyl-m-Toluamide 1.08 1.08 1.08

4. Toluene 1.89 1.88 1.89

1

2 3 4

1

23 4

0 1

1

2

34

2 3 Min.

Initial

Final Base

Final Acid

10252

Basic Compound Stability at pH1 and pH10

1025010249

0 2 4 6 8 Min.

1 2

3

6

4

5

EPS C18

C18 HiLoad

C18

12

3

4

5

612

3

4

5

6

1. Uracil 2. Pyridine 3. Phenol 4. N,N-diethyl-m-toluamide 5. 4-Butyl Benzoic Acid 6. Toluene

Column:

Mobile Phase: 50mM KH2PO4, pH 3.0:CH3CN (40:60)

Column Temp: Ambient (22°C)


Injection Vol.: 5µL


Acid, Base, and Neutral Test Mix

0 1 2 3 4 5 Min.

1

2

3

4

5

1. Toluene2. Hydantoin3. Uracil4. Caffeine5. Allantoin

10461

Column:

(Part No. 86464)

Mobile Phase: Acetonitrile:0.10M Ammonium Formate,

pH 3.0 (95:5)



Mixed Analyte HILIC

miscella

neous


hplc applications

® Alltima HP C18, 5µm, 4.6 x 50mm®Alltech

® Alltima HP , 4.6 x 150mm®Alltech ® Alltima HP HILIC, 5µm, 4.6 x 150mm®Alltech

0 2

2

1

4 6 Min.


Mobile Phase: 0.025M K2HPO4, pH7.0:Tetrahydrofuran:

Methanol (50:25:25)



8584

1. 2,2’-Dipyridyl

2. 8-Hydroxyquinoline

Chelators

Metal Chelators

10429


(Part No. 87685)

Mobile Phase: Acetonitrile:Dichloromethane (60:40)


Detector: ELSD

Paraffin Analysis

0 2 4 6 8 10 12 14 16 18 20 22 Min.

Paraffin C20-C30

0 2 4 6 Min.

1

4

5

6

2+3

C18-HL

2

3

1

4

5

6

C18

1

2

5

4

6

C18-P

3

C18-B

1

3

2

4

5+6

VisionHT

1. Uracil 2. Pyridine 3. Phenol 4. Methyl Alanine 5. 4-Butyl Benzoic Acid 6. Toluene

Column: 2.0 x 50mm

Mobile Phase: 50mM KH2PO4, pH 7.0:CH3CN (60:40)


Standards

miscellaneous

63

hplc applications


Paraffins

Reversed-Phase Comparison®

® ®

0 4 8 12 16 20 24 Min.

1

2

3

4

5

67

8

9

10

0 2 4 6 8 10 12 14 16 18 20 22 24 26 Min.

1

2

3

4 5

6 7

8

9 10 11

1

2

3

0 2 4 6 8 10 12 14 16 18 20 22 24 Min.

0 2

3

2

1

4

4

6 8 10 Min.

0 5

1

10 Min.

2

ic applications

Anions

10120

1. Fluoride 2. Chlorite 3. Bromate 4. Chloride 5. Nitrite 6. Bromide 7. Chlorate 8. Nitrate 9. Phosphate 10. Sulfate

1. Fluoride 2. Chlorite 3. Bromate 4. Chloride 5. Nitrite-N 6. Bromide 7. Chlorate 8. Dichloroacetate 9. Nitrate-N 10. Phosphate-P 11. Sulfate

10121

Anions in Reagent Grade Water

by EPA Method 300.1Anions in High Ionic Strength Water by

EPA Method 300.1

1. Chlorite 2. Bromate 3. Nitrite

10123

Trace Bromate with Post Column Derivatization

and UV Detection, EPA Method 317.0

1. Aspartame 2. Chloride 3. Nitrate 4. Sulfate

1. Fluoride 2. Chloride 3. Phosphate 4. Monofluorophosphate

8458 8464

1. Arsenite 2. Arsenate

9178

Diet Cola Toothpaste

Arsenite and Arsenate

Column: Alltech® Novosep™ A-2 Anion, 250 x 4mm (Part No. 51410)

Mobile Phase: 3.6mM Sodium Carbonate


Column Temp: 43°C

Detector: Suppressed Conductivity




Column Temp: 43°C





Column Temp: 40°C

Reagent: o-Dianisidine


Column: Alltech® Allsep™ Anion, 100 x 4.6mm (Part No. 51207)

Mobile Phase: 4mM Phthalic Acid, pH4.2 with Lithium Hydroxide


Detector: Conductivity


Mobile Phase: 0.85mM Sodium Bicarbonate/0.9mM Sodium Carbonate




Mobile Phase: 4mM p-Hydroxybenzoic Acid, pH8.5

with Lithium Hydroxide




anions

Anions


ic applicationsanions

0 2

1

2

3

45

6

4 6 8 10 Min.

1

2

3

4

5

6

7

0 2 4 6 8 10 12 14 16 18 20 22 24 Min. 0 1 2 3 4 5 6 7 8 9 Min.

1

2

1. Fluoride 2. Chloride 3. Nitrite 4. Bromide 5. Nitrate 6. Phosphate 7. Sulfate

1. Fluoride 2. Chloride 3. Nitrite 4. Bromide 5. Nitrate 6. Sulfate

1. Chloride 2. Nitrate 3. Phosphate 4. Sulfate 5. Oxalate

13033

84328451

Anions

Anions with Phthalic Acid Mobile Phase Oxalate in Urine

13040

1. Iodate 2. Bromate

Bakery Additives

8610 1. Fluoride 7. Selenite

ungstate Molybdate

Selenite, Selenate, Arsenate,

Tungstate and Molybdate










Mobile Phase: 4mM Phthalic Acid, pH4.2 with Lithium Hydroxide


Column Temp: 35°C



Mobile Phase: 0.85mM NaHCO3/ 0.9mM NaHCO3







0

2

1

3

4 56

78

910

11 12

10 20 30 40 Min.

2. Chloride 3. Nitrite 4. Bromide 5. Nitrate 6. Phosphate

8. Sulfate 9. Arsenate 10. Selenate 11. T 12.

Column: Alltech® Anion/R, 10µm, 250 x 4.1mm (Part No. 269029)

Mobile Phase: 4mM Sodium Hydroxide/0.1mM Sodium Benzoate


1. Borate 2. Silicate 3. Formate 4. Sulfide 5. Chloride 6. Cyanide

5697

Weak Acid Anions1

23

4

5

6

0 9 18 Min.

Column: Alltech® Sarasep™ AN1™, 9µm, 250 x 4.6mm

(Part No. 38140)

Mobile Phase: 0.75mM Sodium Bicarbonate, 2.2mM Sodium Carbonate



Fluoride Chloride Nitrite Bromide

Nitrate SelenitePhosphate

Sulfate Selenate

Column: Alltech® ICSep™ AN1™, 9µm, 250 x 4.6mm

(Part No. 38140)

Mobile Phase: 1.8mM Sodium Bicarbonate, 1.8mM Sodium Carbonate



FluorideChloride

Bromide Nitrate Phosphate Sulfite

Sulfate

8611 6316

Selenite, Selenate, and Other Anions Separation of Sulfite and Sulfate

1. Fluoride 2. Bicarbonate 3. Chloride 4. Nitrite 5. Bromide 6. Nitrate 7. Phosphate 8. Sulfate

Column: Alltech® Anion/R, 10µm, 250 x 4.1mm (Part No. 269029)

Mobile Phase: 5mM p-Hydroxybenzoic Acid, pH8.5

with Lithium Hydroixde



6159Anion Standards

13038

Column: Alltech® OA-1000, 9µm, 300 x 6.5mm (Part No. 9046)

Mobile Phase: 1.0% Phosphoric Acid


Column Temp: 35°C

Detector: Conductivity (Non-Suppressed)

Haloacetic Acids Anions

1

2

3

4

5

6

7

8

0 9 18 Min. 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.

1

2

1. Chloro-difluoro Acetic Acid 2. Difluoro Acetic Acid

0 4

1

23

4 5

6

7

8

9

8 12 16 Min.

1. 2. 3. 4. 5. 6. 7. 8. 9.

1

2

34

5 6

7

8

9

0 2 4 6 8 10 12 14 16 Min.

1. Water 2. 3. 4. Nitrite 5. 6. 7. 8. 9.

0 4 8 12 16 20 24 Min.

1

2

3 4

1. Nitrite 2. EDTA 3. Bromide 4. Nitrate

Column: Anion/S, 10µm, 4.6 x 100mm (Part No. 269012)

Mobile Phase: 1.5mM Sulfuric Acid



EDTA and Other Anions

8284


anions

ic applications


cations

ic applications

0 5

1

2

3

4

5

10 15 20 Min.

8462

Column: Alltech® Universal Cation, 7µm, 100 x 4.6mm

(Part No. 27106)

Mobile Phase: 2.5mM Oxalic Acid



1. Sodium 2. Ammonium 3. Potassium 4. Magnesium 5. Calcium

Trace Level Ammonium in Brine

1

2

3

4

5

6

78

0 10 20 30 40 Min.

0 4

1

23

4 5 6

7

8 12 16 Min.

13013

6292

Column: Alltech® Universal Cation HR, 3µm, 53 x 7.0mm

(Part No. 23109)

Mobile Phase: 3mM Methanesulfonic Acid



1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Magnesium 6. Calcium


(Part No. 27106)

Mobile Phase: 5mM Hydrochloric Acid



1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Rubidium 6. Magnesium 7. Calcium 8. Barium

Rapid Analysis of Monovalent

and Divalent Cations

Rubidium and Barium


(Part No. 27106)




1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Morpholine 6. Magnesium 7. Calcium


(Part No. 27106)




1. Monoethanolamine 2. Diethanolamine 3. Triethanolamine 4. Magnesium 5. Calcium

8632

6315

Morpholine and Group I & II Cations

Ethanolamines

Cations

1

2

3

45

0 1 2 3 4 5 Min.

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Min.

1

23

4

5

6

7

8

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 Min.

1

2

3

4

5 67

89

0 2 4 6 8 10 12 14 16 18 20 Min.

1

23

4

5

6

78


(Part No. 27106)




1. Potassium 2. Magnesium 3. Calcium

Column: Hamilton® PRP™-X200, 10µm, 150 x 4.1mm

(Part No. 79441)

Mobile Phase: 4mM Nitric Acid in 30% Methanol



1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Cesium

6303 5204

Cations in Coffee Extract Alkali Metals

13035


(Part No. 27106)

Mobile Phase: 2mM Tartaric Acid/1mM Oxalic Acid



1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Nickel 6. Zinc 7. Cobalt 8. Magnesium 9. Calcium


(Part No. 27106)




1. Lithium 2. Sodium 3. Ammonium 4. Potassium 5. Cesium 6. Magnesium 7. Calcium 8. Strontium


(Part No. 27106)

Mobile Phase: 3mM Tartaric Acid/2mM Oxalic Acid



1. Copper 2. Sodium 3. Ammonium 4. Potassium 5. Zinc 6. Lead 7. Magnesium 8. Calcium

10318 13042

Metal Ions

Cesium and Strontium

Transition Metals

Cations


catio

ns

ic applications

1

2

18 Min.0 4 8 12 14 162 6 10

0 2 4 6 8 10 12 14 16 18 20 Min.

1

2 3

4

Before SPE Clean-up

12

3

4

After SPE Clean-up

SPE Extraction of Salicylic and Acetylsalicylic Acids from Synthetic Urine

Column: Alltima HP C18 HiLoad, 5µm, 250 x 4.6mm

(Part No. 87698)

Mobile Phase: 5mM Sodium Acetate, pH5.2:Methanol (80:20)



1. Salicylic Acid 2. Acetylsalicylic Acid

10642

1. Oxazepam 2. Clonazepam 3. Temazepam 4. Diazepam

Benzodiazepines from Human Plasma

10606

10608

Analgesics

Sedatives

Procedure using GracePure™ C18-Aq, 500mg:

Sample Treatment – Spike 500µL plasma with 10µL of a 0.1mg/mL standard solution.

Combine with 100µL of 0.1M sodium carbonate buffer. Vortex and centrifuge for

10 minutes.

Conditioning – Rinse device 2mL water.

Sample Application – Apply entire sample.

Wash – Wash with 2mL water and then 50µL methanol.

Elution – Elute with 600µL methanol.

Column: Alltima HP C18, 5µm, 250 x 4.6mm (Part No. 87680)



%B: 30 80

Flow Rate: 1mL/min


Procedure using GracePure™ C18-Max, 100mg:

Sample Treatment – Spike 2mL synthetic urine with 100ppm salicylic acid and 100ppm

acetylsalicylic acid.

Conditioning – Rinse device with 3mL methanol followed by 3mL DI water.

Sample Application – Apply 2mL spike urine.

Wash – Wash with 2mL 50mM phosphate buffer monobasic, pH 2.

Elution – Elute with 2mL methanol:water (50:50).


spe applicationsp

ha

rma

ce

uti

ca

ls

®

®

0 1 2 3 4 5 6 7 8 9 10 Min.

12

3

4

5

6

78

After SPE

Concentration

Before SPE

Concentration

0 1

1

2

2

3 Min.

0 1

1

2 3 Min.

spe applications

ww.discoverysciences.com

nutra

ceutic

als

1. Acetazolamide 2. Hydrochlorothiazide 3. Hydroflumethiazide 4. Clopamide 5. Trichloromethiazide 6. Indapamide 7. Bendroflumethiazide 8. Spironolactone

Diuretics from Urine

10289

10290

Diuretics

Column: Alltima™ C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)

Mobile Phase: Methanol:0.2% Phosphoric Acid (95:5)


Detector: VIS at 585nm

1. Pseudohypericin 2. Hypericin

9107

Hypericins from St. John’s Wort

Column: Alltima™ C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)

Mobile Phase: Acetonitrile:20mM K2HPO4, pH7.3 (65:35)

Flow Rate: 3mL/min

Detector: UV 340nm

Inj. Vol.: 5µL

1. Apigenin-7-Glucoside

9133

Antioxidants from Chamomile

Procedure Using GracePure™ C18-Max, 1000mg:

Sample Treatment – Pulverize 300mg St. John’s Wort powder into five, 3mL aliquots

of methanol:water (80:20). Combine and filter extracts to produce 15mL filtrate. Dilute

to 30mL with water.

Conditioning – Rinse device with 5mL methanol followed by 5mL deionized water.

Sample Application – Apply 2mL filtrate to the top of the SPE device and draw through

the C18 packed bed.

Wash* – Wash device with 3mL of deionized water.

Elution – Elute with 2mL methanol.*Repeat load and wash steps consecutively until filtrate is consumed.


Sample Treatment – Pulverize 1g commercial chamomile tea grounds into

6mL dioxane:methanol (50:50). Filter extract and dilute to 20mL with water.

Conditioning – Rinse device with 5mL methanol followed by 5mL water.

Sample Application* – Apply 1mL filtrate.

Wash* – Wash with 1mL water.

Elution – Elute first with 2mL methanol:water (50:50). Elute second, fraction

and use for this analysis, with 2mL methanol.*Before elution step, repeat load and wash steps until filtrate is consumed.

St. John’s Wort


Sample Treatment – Spike synthetic urine with 8 diuretics to a concentration of

1.25µg/mL each.

Conditioning – Rinse device 5mL methanol, followed by 5mL water.

Sample Application – Apply 15mL spiked urine sample at 1mL/min.

Wash – Wash with 5mL water.

Elution – Elute with 2mL methanol. Evaporate solvent and reconstitute in 250µL

HPLC mobile phase.

Chamomile

Column: Alltima™ C18, 3µm, 100 x 4.6mm (Part No. 81382)

Mobile Phase: A: 25mM Ammonium Acetate, 0.1%Trifluoroacetic Acid

B: Acetonitrile, 0.1%Trifluoroacetic Acid


%B: 20 90


Detector: ELSD

0 1

1

2

2

3

3

4

4

5

5

0 1

1

2 3 4 5 Min.

0 1

1

2

3

2 Min.

Column: Alltima C18, 3µm, 53 x 7mm Rocket™ (Part No. 50605)

Mobile Phase: Acetonitrile:Isopropyl Alcohol:0.2% Acetic Acid (17:23:60)


Detector: UV 220nm

Inj. Vol.: 5µL

1. Methysticin 2. Yangonin 3. Kavain 4. Dihydrokavain 5. Dihydromethysticin

9125

Sedatives from Kava Kava

Column: Alltima C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)

Mobile Phase: Methanol:0.2% Acetic Acid (65:35)


Detector: UV 270nm

Inj. Vol.: 5µL

1. Z-ligustilide

9132

Vasodilators from Dong Quai

Column: Alltima C18, 3µm 53 x 7mm Rocket™ (Part No. 50605)

Mobile Phase: Acetonitrile:10mM K2HPO4, pH2.6 (20:80)



1. Echinicoside 2. Chlorogenic Acid 3. Caffeic Acid

9110

Phenolic Acids from Echinacea


Sample Treatment – Pulverize 300mg echinacea powder into five, 3mL aliquots of

methanol. Combine and filter extracts to produce 15mL filtrate. Dilute to 30mL with

water.

Conditioning – Rinse device with 5mL methanol followed by 5mL deionized water.

Sample Application – Apply 2mL filtrate to the top of the SPE device and draw through

the C18 packed bed.

Wash* – Wash device with 2mL of deionized water.

Elution – Elute with 5mL of methanol:water (50:50).*Repeat load and wash steps consecutively until filtrate is consumed.


Sample Treatment – Pulverize 300mg commercial kava kava root powder into four, 2mL

aliquots of methanol:water (80:20). Combine and filter extracts to produce 8mL filtrate.

Dilute to 10mL with water.




Elution – Elute with 3mL methanol.*Before elution step, repeat load and wash steps until filtrate is consumed.

Echinacea

Kava Kava

Dong Quai


Sample Treatment – Pulverize 300mg commercial kava kava root powder into four, 2mL

aliquots of methanol:water (80:20). Combine and filter extracts to produce 8mL filtrate.

Dilute to 10mL with water.




Elution – Elute with 3mL methanol.*Before elution step, repeat load and wash steps until filtrate is consumed.


spe applicationsn

utr

ace

uti

ca

ls

®

®

®

0 10 20 30 35 40 45 50 605 15 25 55 65 Min.

Before SPE Clean-up

1

2

6

34

5

After SPE Clean-up

Column: Heliflex® AT™ AquaWax-DA, 30m x 0.25mm x 0.25µm

Capillary GC Column (Part No. 14537)

Temp: 200°C (5 min hold) to 230°C (4 min hold) at 5°C/min

Carrier Gas: Helium at 0.75mL/min (25cm/sec)

Detector: FID at 250°C

1. Sorbic Acid 2. Benzoic Acid

2576

Preservatives from Fruit Punch

Additives

Procedure using GracePure™ Anion-X, 500mg:

Sample Treatment – Use 8mL fruit punch and adjust pH to 10 using potassium

hydroxide.

Conditioning – Rinse device 10mL water.

Sample Application – Apply 8mL pH adjusted fruit punch sample.

Wash – Wash with 20mL water.

Elution – Elute with 1mL 1.0N hydrochloric Acid followed by 1mL methanol.

1. Carbendazim 2. Thiabendazole 3. Triadimenol 4. Triadimefon 5. Flusilazole 6. Diniconazole

Fungicides from Red Wine

10637

10638

Carbohydrates


Sample Treatment – Add 0.167mg/mL each of carbendazim, thiabendazolem,

triadimenol, triadimefon, flusilazole, diniconazole into

1mL water. Combine with 1mL Beaujolais red wine.


Sample Application – Apply 2mL red wine mixture.

Wash – No wash.

Elution – Elute with 3mL methanol.

Column: Alltima HP C18 Amide, 5µm, 250 x 4.6mm

(Part No. 87734)



%B: 15 45 45 15

Flow Rate: 1mL/min



spe applicationsfo

od

& b

eve

rag

e

®

Melamine

0 2 4 6 10 128 14 Min.

0

40

80

120

160

mAU

Melamine from Biscuit

SPE Column Type: ™ 5138770)

Sample Pretreatment: weight biscuit 2g

melamine standard solution

Mobile Phase:

Column: ®

Detector:

Flow Rate:

Column Temperature:

Injection:

SPE Column Type: ™ 5138770)

Sample Pretreatment: weight pork 2g


SPE recovery*=103%

HPLC Mobile Phase:

Column: ®

Detector:

Flow Rate:

Column Temperature:

Injection:

SPE Steps:

Column Conditioning: 2 column volumes methanol, then 2 column volumes water

Sample Application:

Column Washing: 3 column volumes water, then 2 column volumes methanol

Eluent:

Melamine standard

Spiked pork before SPE

Spiked pork after SPE

SPE Steps:


Sample Application:


Eluent:

Melamine from Pork

0 2 4 6 10 128 14 Min.

0

40

80

120

160

mAU

SPE recovery*=80%

Melamine standard

Spiked biscuit before SPE

Spiked biscuit after SPE

0 2 4 6 10 128 14 Min.

0

50

100

150

200

mAU

Melamine from HamSPE Column Type: ™ 5138770)

Sample Pretreatment: weight ham 2g


SPE recovery*=92%

Mobile Phase:

HPLC Column:

Detector:

Flow Rate:

Column Temperature:

Injection:

SPE Steps:


Sample Application:


Eluent:

Melamine standard

Spiked ham before SPE

Spiked ham after SPE


spe applicationsfo

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& b

eve

rag

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®

1

2

3

0 1 2 3 4 5 6 7 8 9 Min.

1. Chloride 2. Ribonuclease A 3. Cytochrome C

10688

Prevail™ SPE Protein Desalting, Cytochrome C and Ribonuclease A Mix in 10mM Sodium Chloride

Protein Extraction of Ribonuclease and Myoglobin

1. Ribonuclease 2. Myoglobin

Proteins & Peptides/Reversed Phase

Procedure using Vydac® SPE:

A 3mL SPE cartridge was conditioned with 1mL of Acetonitrile followed by 1mL of

5% Acetonitrile/0.1% Trifluoroacetic Acid. Ribonuclease and myglobin (100mg each)

were then loaded in 30% Acetonitrile/0.1% Trifluoroacetic Acid . The cartridge was

washed with 1mL of 5% Acetonitrile/0.1% Trifluoroacetic Acid to remove weakly bound

compounds, then 1mL of 30% Acetonitrile/0.1% Trifluoroacetic Acid followed by 1mL of

60% Acetonitrile/0.1% Trifluoroacetic Acid. HPLC analysis of the 5% Acetonitrile wash

(A) revealed only a small amount of ribonuclease. Most of the ribonuclease eluted in the

30% Acetonitrile wash (B). The myoglobin eluted almost entirely in the 60% Acetonitrile

wash (C).

Procedure using Prevail™ C18, 500mg:

Sample Treatment – Cytochrome-C 1mg/mL, Ribonuclease-a 1mg/mL, Blank = water,

Salt = 10mM NaCl.

Conditioning – 1 tube volume of methanol (do not dry). 2 tube volumes of 0.025%

ammonium hydroxide. Dry tube via vacuum.

Sample Application – Pipette 1mL of sample (either proteins in water blank or proteins

in salt solution) onto top of sorbent bed in SPE tube. Run vacuum @ approximately

-10 kPa, loading sample in a tight band at top of sorbent column, continue to apply

vacuum (@ -10 kPa) until tube is dry. Larger (2mL +) volumes can be de-salted and

concentrated. Liquid collected will contain the NaCl

Elution – Pipette 500µL 0.4% TFA and 500µL Acetonitrile containing 0.4% TFA onto top

of sorbent bed in spe tube. Run vacuum @ approximately -10 kPa, eluting sample from

sorbent column, continue to apply vacuum until tube is dry.

Column: ProSphere™ 300 C4-300, 5µm, 150 x 4.6mm

(Part No. 33989)

Mobile Phase: A: 0.2% Trifluoroacetic Acid B: Acetonitrile


%B: 25 70

Flow Rate: 1mL/min

Detector: ELSD


Mobile Phase: A: 0.1% Trifluoroacetic Acid B: Acetonitrile


%B: 15 70

spe applicationslife

scie

nce

s


Before SPE Clean-up

After SPE Clean-up

0 10 20 305 15 25 35 Min.

123

45

6

7 8

9101211

13

15

14

16

Before SPE Clean-up

After SPE Clean-up

0 10 20 305 15 25 35 Min.

12

3

4

5 6

78

109 1112

131415+16

After SPE Clean-up

0 5 10 15 20 25 30 35 40 Min.

Column: AT™ 5ms, 0.25µm, 30m x 0.25mm (Part No. 15807)

Temp: 160°C (0 min hold) at 10°C/min, then to 275°C at 5°C/min


Detector: ECD at 310°C

Chlorinated Pesticides in Vegetables2780

2789

Chlorinated Pesticides in Soil

2779

2788

1. Aroclor 1254

PCBs in Transformer Oil


Temp: 160°C (0 min hold) at 10°C/min, then to 275°C at 5°C/min







2784

Pesticides

Procedure using GracePure™ Florisil®, 1000mg:

Sample Treatment – Dissolve 0.25grams of transformer oil spiked with Aroclor 1254 (at

a concentration of 50mg/kg) in 20mL n-hexane.

Conditioning – 2 x 5mL n-hexane, making sure the cartridge does not dry.

Sample Application – Attach 25mL reservoir (Part No. 210425) with a syringe adapter

(Part No. 210705) and run sample through conditioned Florisil® tube, aspirating all the

solution from the tube. Evaporate the volume down to 4mL and analyzed by GC/ECD.

Wash – No wash.

Elution – No elution, collect sample load.


Sample Treatment – Homogenize 5 grams green bell pepper with 25mL methanol, spike

with 0.5µg each of 16 chlorinated pesticides. Filter with 75mL filter column (Part No.

210775). Dilute to 200mL with DI water and shake.

Conditioning – 2 x 5mL methanol, then 5mL DI water.


(Part No. 210705) and apply entire sample.

Wash – No wash, apply vacuum for 5 min.

Elution – Elute chlorinated pesticides with 2 x 1mL n-hexane/CH2Cl2 (70:30),

evaporated this to dryness and reconstituted with 1mL n-hexane.


Sample Treatment – Weighed out 5 grams dry soil, spike with 1µg each of 16

chlorinated pesticides, add added 25mL methanol and homogenize. Filter with 75mL

filter column (Part No. 210775). Dilute to 200mL with DI water and shake.

Conditioning – 2 x 5mL methanol, then 5mL DI water.


(Part No. 210705) and apply entire sample.

Wash – No wash.

Elution – Elute chlorinated pesticides with 2 x 1mL n-hexane/CH2Cl2 (70:30), evaporate

to dryness and reconstitute with 1mL iso octane.

1. -BHC 2. -BHC 3. -BHC 4. -BHC

5. Heptachlor 6. Aldrin 7. Heptachlor Epoxide 8. Endosulfan I 9. p,p’-DDE

10. Dieldrin 11. Endrin 12. Endosulfan II 13. p,p’-DDD 14. Endrin Aldehyde 15. p,p’-DDT 16. Endosulfan Sulfate

1. -BHC 2. -BHC 3. -BHC 4. -BHC 5. Heptachlor

6. Aldrin 7. Heptachlor Epoxide 8. Endosulfan I 9. p,p’-DDE 10. Dieldrin

11. Endrin 12. Endosulfan II 13. p,p’-DDD 14. Endrin Aldehyde 15. p,p’-DDT 16. Endosulfan Sulfate


spe applicationsenvironmental

Before SPE Clean-up

After SPE Clean-up

0 10 20 305 3515 25 40 Min.

0 9 18 Min.

12

3

45

6

7

89

10

11

12

13

14

15

16 17

18

0 5 10 15 20 25 Min.

Before SPE

Concentration

6

5

4

321

After SPE

Concentration

1. Aroclor 1242

PCBs in Salmon

2781

2787





Pesticides

Procedure using GracePure™ C18-Fast, 1000mg:

Sample Treatment – Weigh out 2.5 grams salmon, spike with 2.5µg Aroclor 1242, add

25mL acetonitrile, homogenize for 30 seconds. Filter with 75mL filter tube (Part No.

210775). Rinse salmon again with 5mL acetonitrile. Bring up to 100mL with deionized

water.

Conditioning – Rinse with 6mL hexanes, 6mL acetone, 2 x 6mL methanol, 2 x 6mL

water.

Sample Application – Attach 75mL reservoir, apply entire sample.

Wash – 2 x 5mL water. Dry columns for 10 minutes at full vacuum.

Elution – Elute PCBs with 2 x 3mL 3% toluene/97% n-hexane. Evaporate

solvent and reconstitute in 1mL n-hexane.

Chlorinated Pesticides from Water

Column: AT™ Pesticide 20m x 0.53mm x 0.60µm Capillary GC

Column (Part No. 16846)

Temp: 210°C

Carrier Gas: Helium, 35cm/sec

Detector: ECD


Conditioning – Rinse device with 5mL of methanol followed by 5mL deionized water.

Sample Application – Pass 100mL–500mL (containing 1% methanol) of water sample

through the device at 20mL/minute.

Wash – Wash device with 10mL of deionized water then 10mL of methanol:deionized

water (20:80). Remove excess by passing air through the device for two minutes.

Elution – Elute with 2mL of hexane:ethyl acetate (70:30). Pass extract through 2g–3g

sodium sulfate to remove residual water.

1668

Recovery (as % Standard) 1. -BHC (104%) 2. Lindane (109%) 3. -BHC (106%) 4. Heptachlor (92%) 5. -BHC (110%) 6. Aldrin (98%) 7. Heptachlor Epoxide (107%) 8. -Endosulfan (105%) 9. p,p'-DDE (79%) 10. Dieldrin (102%) 11. Endrin (104%) 12. p,p'-DDD (92%) 13. -Endosulfan (103%) 14. p,p'-DDT 15. Endrin Aldehyde (102%) 16. Endosulfan Sulfate (103%) 17. Methoxychlor (99%) 18. Endrin Ketone (104%)

Carbamate Pesticides from Water


Sample Treatment – Spike 500mL tap water with 125µL carbamate solution for final

concentration of 25ppb.

Conditioning – Rinse with 3mL acetonitrile:water (80:20) followed by 3mL water. Dry

with vacuum.

Sample Application – Apply 500µL sample.

Wash – 2 x 3mL water.

Elution – Elute with 4 x 1mL acetonitrile:water (80:20)

10599

10600

1. Aldicarb sulfoxide 2. Methomyl 3. 3-Hydroxycarbofuran 4. Propoxur 5. Carbaryl 6. Methiocarb

Column: Platinum™ EPS C18, 5µm, 250 x 4.6mm (Part No. 32246)

Mobile Phase: A: DI water B: Acetonitrile


%B: 25 25 50 50 25

Flow Rate: 1mL/min



enviro

nmental

spe applications

Before SPE Clean-up

After SPE Clean-up

0 10 20 30 35 405 15 25 45 50 Min.

1

23

4

56 7 8

910

11

1213

151416

Before SPE Clean-up

After SPE Clean-up

0 10 20 30 35 405 15 25 45 50 Min.

1

23

456 7

8

109 1211 13

1514

16

PAHs in Water2771

2772

PAHs in Toasted Oat Cereal

2775

2791


9. Benzo(a)anthracene 10. Chrysene 11. Benzo(b)fluoranthene 12. Benzo(k)fluoranthene 13. Benzo(a)pyrene 14. Indeno(1,2,3-cd)pyrene 15. Dibenzo(a,h)anthracene 16. Benzo(ghi)perylene


Temp: 80°C (1 min hold) to 160°C (0 min hold) at 25°C/min,

then to 300°C (0 min hold) at 3°C/min, then to 325°C

(3 min hold) at 25°C/min

Carrier Gas: Helium at 1mL/min (28cm/sec)










PAHs

Procedure using GracePure™ C18-Max:

Sample Treatment – 500mL of water and add 75mL of 2-propanol,

spiked with 20uL of PAH mixture at 2000ug/mL of each PAH, which corresponds to

40ug/500mL water, or 80ppb of each PAH.

Conditioning – 2 x 5mL of methanol, then 5mL of 85% water/

15% 2-propanol.

Sample Application – Attach 75mL reservoir (Part No. 210575) with a

syringe adapter (Part No. 210705) and apply entire sample. Dry

columns for 30 minutes at full vacuum.

Wash – No wash.

Elution – Elute PAHs with 3 x 1mL CH2Cl2 and adjusted the volume

to 1mL by evaporating under a gentle nitrogen stream.

Procedure using GracePure™ C18-Max:

Sample Treatment – 500mL of water and add 75mL of 2-propanol, spiked with 20uL of

PAH mixture at 2000ug/mL of each PAH, which corresponds to 40ug/500mL water, or

80ppb of each PAH.

Conditioning – 2 x 5mL of methanol, then 5mL of 85% water/15% 2-propanol.


(Part No. 210705) and apply entire sample. Dry columns for 30 minutes at full vacuum.

Wash – 2 x 500µL of 85% water/15% isopropyl alcohol. Vacuum 10 min.

Elution – Elute PAHs with 3 x 1mL CH2Cl2 and adjusted the volume to 1mL by

evaporating under a gentle nitrogen stream.


environmental

spe applications

0 5 10 15 20 25 30 35 Min.

Before SPE

Concentration

1

2

3

4

56

After SPE

Concentration

0 5 10 15 20 25 30 35 40 45 50 55 Min.

1

2

3 4 56 7

89 10

1112

13

14

1516

Before SPE

Concentration

After SPE

Concentration

PAHs in Cooking Oil2776

2792









PAHs

Nitroaromatics

10597

10598

Nitroaromatics and Naphthols from Soil

Column: Adsorbosphere™ UHS C18, 5µm, 150m x 4.6mm

(Part No. 288118)





Sample Treatment – Spike 100g soil with 7.5µg/g of each analyte. Combine soil with

1000mL deionized water, shake for 10 minutes, and filter.


Sample Application – Aspirate 1000mL water sample through SPE at flow rate 1-

5mL/min.

Wash – No wash. Air dry for 15 seconds.

Elution – Elute with three 1mL aliquots of methanol:water (50:50). Air dry

for 15 seconds between each elution.

1. 2,4-Dinitrotoluene 2. 1-Naphthol 3. 2-Naphthol 4. 2-Nitrotoluene 5. 4-Nitrotoluene 6. 3-Nitrotoluene

Procedure using DVB Extract-Clean™, 500mg/4mL:

Sample Treatment – Weigh out 1 grams canola oil, spike with 20µg of each PAH, add

10mL n-hexane.

Conditioning – 10mL toluene, applied vacuum until toluene removed, then 5mL n-hexane.

Sample Application – Aspirate the entire canola oil/hexane/PAH solution through the tube.

Wash – 5mL n-hexane.

Elution – Elute PAHs with 4 X 2.5mL toluene and adjusted the sample volume to 0.5mL

(500µL) by evaporating under dry nitrogen stream and gentle heat.


enviro

nmental

spe applications

0 2 4 6 8 10 12 14 16 18 20 Min.

Before SPE Clean-up

After SPE Clean-up

1.0

1.2

1.4

1.6

1.8

0.0

0.2

0.4

0.6

0.8

2.0

2.2

2.4

2.8

2.6

3.0

1.0

1.2

1.4

1.6

1.8

0.0

0.2

0.4

0.6

0.8

2.0

2.2

2.4

2.8

2.6

3.0

Resp

on

se -

Mil

liV

olt

s

1

2

34

5

1. Simazine

2. Atrazine

3. Propazine

4. Terbuthylazine

5. Cyanazine

GC Conditions

Injected 1.6uL of above sample onto AT™ 5ms column, 30m x 0.25mm i.d., 0.25μm film thickness (Part #15807) under the following conditions:

Oven: 80°C (0 minutes)-280°C (0 minutes) at 10°C/min

Injector: 250°C

FID: 310°C, 14.5psig helium

Linear Velocity: 29cm/sec

Flow: 1.1mL/minute

Split Ratio: 50:1

SVF: 55mL/minute (all per Agilent 6890)

SPE Conditions

Procedure using GracePure™ C18-Fast columns, 500mg/3mL, (Part #5138758)

Sample Treatment: Spiked 500mL of water

spiked with 10ug each triazine herbicide

(simazine, atrazine, propazine, terbuthylazine,

and cyanazine).

Conditioning: 2 x 2mL acetone, 2 x 2mL

methanol, 2 x 2mL water. Column must not

run dry!

Sample Application: Attached a 75mL

reservoir (Part #210575) to the SPE tube with

a syringe adapter (Part # 210705) and aspirate the sample through the tube.

Elution: Elute triazine herbicides with 3 x 1mL

acetone. Adjust sample volume to 1mL by evaporating off acetone with a warm nitrogen

stream.

Triazine Herbicides in Water

Triazine


environmental

spe applications

GRACE® is a trademark, registered in the United States and /or other countries, of W. R. Grace & Co.-Conn.

GRACEPURE™ and GRACESMART™ are trademarks, registered in the United States and / or other countries, of

W. R. Grace & Co.-Conn. ALLTECH®, ALLTIMA®, DENALI®, EVEREST®,GENESIS®, HELIFLEX®,

VISIONHT® and VYDAC® are trademarks, registered in the United States and/or other countries,

of Alltech Associates, Inc. ADSORBOSPHERE™, ALLSEP™, ALLSPHERE™, AT™, EXPEDITE™,

EXTRACT-CLEAN™, NOVOSEP™, PLATINUM™, PREVAIL™, PROSPHERE™, PROZAP™ and ROCKET™

are trademarks, registered in the United States and/or other countries of Alltech Associates, Inc.

ASTECH™ is a trademark, registered in the United States and/or other countries, of ATEX Explosionschutz GmbH.

CHIROBIOTIC® is a trademark, registered in the United States and/or other countries, of Sigma-Aldrich

Biotechnology LP.

Conquistador® is a trademark, registered in the United States and/or other countries, of Heaven Hill Distilleries, Inc.

FLORISIL® is a trademark, registered in the United States and/or other countries, of U.S. Silica Company.

Hamilton® is a trademark, registered in the United States and/or other countries, of Hamilton Company.

ICSep™ is a trademark, registered in the United States and/or other countries, of Transgenomic, Inc.

Kellogg’s Frosted Flakes®, is a trademark, registered in the United States and/or other countries, of Kellogg North

America Company.

Pepsi One® is a trademark, registered in the United States and/or other countries, of PepsiCo, Inc.

ROBITUSSIN® is a trademark, registered in the United States and/or other countries, of Wyeth LLC.

SaraSep™ is a trademark, registered in the United States and/or other countries, OF Transgenomic, Inc.

SPLENDA® is a trademark, registered in the United States and/or other countries, of McNeil Nutritionals, Inc.

Taxol® is a trademark, registered in the United States and/or other countries, of Bristol-Myers Squibb Company.

ZirChrom® is a trademark, registered in the United States and/or other countries, of Zirchrom Separations, Inc.

This catalog is an independent publication and is not affiliated with, nor has it been authorized, sponsored,

or otherwise approved by any of the companies referenced above. This trademark list has been compiled using

available published information as of the publication date of this brochure and may not accurately reflect current

trademark ownership or status. ©Copyright 2015 Alltech Associates, Inc. All rights reserved.

grace.com

Global Headquarters W. R. Grace & Co.-Conn7500 Grace DriveColumbia, Maryland 21044 USA

North America2051 Waukegan RoadDeerfield, IL 60015United StatesTel: +1 847 948 8600Fax: +1 847 948 1078Email: [email protected]

China19th Floor, K. Wah Center1010 Huai Hai Zhong RoadShanghai 200031 ChinaTel: +86 21 31582888 Email: [email protected]

Hong KongUnits 1001-3, 10 / F AXA Centre151 Gloucester RoadWanchai Hong KongTel: +852 2590.2729Fax: +852 2856.0712

JapanKohken New River Bldg 3F2-21-18, ShinkawaChuo-ku, Tokyo 104-0033 JapanTel: +81 3.3537.6006 Fax: +81 3.3537.6007

BelgiumBrandstraat 12 B-9160 Lokeren BelgiumTel: +32 (0)9 340 65 65 Fax: +32 (0)9 340 65 60 Email: [email protected]

The NetherlandsPB 6596 NL-4802 HN BredaNetherlandsTel: +31 (0)76 571 75 76 Fax: +31 (0)76 571 46 56Email: [email protected]

FranceBâtiment DS - 31 Route de Gallardon – BP 3928234 EpernonFranceTel: +33 (0) 2 37 18 86 70Fax: +33 (0) 2 37 18 86 69Email: [email protected]

GermanyIn der Hollerhecke 167547 WormsGermanyTel: +49( 0) 6241/403-2037Fax: + 49 (0) 6241/403-8193Email: [email protected]

ItalyVia Trento 720017 Passirana di Rho MIItalyTel: +39 02 935 37 1Fax: +39 02 935 37 650Email: [email protected]

United KingdomUnit 6 Boundary WayKellet Road Industrial EstateCarnforth, LA5 9XP, UKTel: +44 (0)1524 734451 Fax: +44 (0)1524 733599Email: [email protected]

Russian Federation18, Yamskogo Polya 3rd StreetMoscow, 125040RussiaTel: +7 495 9374839Email: [email protected]

Australia40 Scanlon DriveEpping Vic 3076AustraliaTel: +61 3 8401 6369Fax: + 61 3 8401 6301 Email: [email protected]

New ZealandPO Box 100352 North Shore Mail CentreNorth Shore City, 0745New ZealandTel: +64 9 447 1614Fax: +64 9 447 1613Email: [email protected]

India17, Commerce Centre, Paud Road, KotharudPune 411 038IndiaTel: +91 20 66449900Email: [email protected]

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