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Application Notes
Aracus
membraPure GmbH
Wolfgang Küntscher Str. 14
16761 Hennigsdorf / Berlin
Mail: [email protected]
Tel.: 03302 / 201 20 0
Fax: 03302 / 201 20 21
___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Index
• Physiological Extended 54 amino acid
Clinical Applications
• Physiological (125x3/iVD) 42 amino acid
• Physiological (150x4)
• Diagnostic short (MSUD)
Food and Feedstuff Applications
• Hydrolysate (125x3)
• Hydrolysate (125x4)
• Hydrolysate PLUS (TRP)
• Feedstuff
• 24 Amino Acids Feedstuff-short
Biogene amine and others
• Biogenic Amine
• Glyphosate and AMPA
• Stress factors plants
Definitions of abbreviations
1 – 2
5 – 6
7 – 8
9 – 10
13 – 14
15 – 16
17 – 18
19 – 20
21 – 22
25 – 26
27 – 28
29 – 30
31 – 32
Seite 1 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Physiological Extended
Equipment .................... : ARACUS
Column ......................... : Lithium system, 150 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent A to Eluent F
Runtime ........................ : 128 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 240 µl/min
Column Temperature ... : 35 °C to 74 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 120 µl/min
Reactor Temperature ... : 125 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Physiological standard Solution
Standard Solution with 100 nmol/ml (Channel A / 570nm)
[min.]Time
0 20 40 60 80 100 120 140
[mV]
Voltage
0
100
200
300
400
500
3,7
3,8
SULFO
6,0
TAU
7,1
PEA
9,6
Ure
a
13,7
GLUA
21,3
ASP
24,5
HYP
28,2
THR
30,3
SER
34,2
ASN
36,5
GLU
39,3
GLN
42,5
SAR
45,6
AAA
48,9
50,8
GLY
53,7
ALA
56,3
CIT
59,5
ABU
65,9
VAL
67,6
SAC
68,0
CYS2
68,6
ASA
69,2
PIP
69,7
CYSTA
70,2
HCI
71,2
MET
71,9
AIL
E
73,5
ILE
75,4
LEU
77,5
NO
RLEU
80,0
TYR
86,3
PHE
91,0
HCY2
95,5
BALA
96,8
ANHYD A
SA
97,2
BAIB
99,5
ALEVA
100,1
GABA
101,2
KYN
102,7
HIS
104,0
1M
HIS
106,2
3M
HIS
108,9
TRP
112,7
CAR
113,7
HCAR 117,1
ANS
119,6
AEC
120,3
HYL
121,8
ORN
123,6
LYS
126,6
Am
monia
128,9
ETN
135,1
ARG
[min.]Time
65 70 75 80
[mV]
Voltage
0
50
100
150
200
65,9
0 V
AL
67,5
8 S
AC
67,9
7 C
YS2
68,5
8 A
SA
69,1
7 P
IP
69,7
0 C
YSTA
70,2
3 H
CI
71,2
1 M
ET
71,9
1 A
ILE
73,5
3 ILE
75,4
4 L
EU
77,4
6 N
ORLEU
80,0
5 T
YR
Seite 2 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Physiological Extended +
number retention time
code
number retention time
code [min]
[min]
1 3,8 SULFO
27 71,2 AILE
2 6 TAU
28 73,5 ILE
3 7,1 PEA
29 75,4 LEU
4 9,6 Urea
30 77,5 NORLE
5 13,7 GLUA
31 80 TYR
6 21,3 ASP
32 86,3 PHE
7 24,5 HYP
34 91 HCY2
8 28,2 THR
35 95,8 BALA
9 30,3 SER
36 96,8 ANHYD ASA
10 34,2 ASN
37 97,2 BAIB
11 36,5 GLU
38 99,5 ALEVA
12 39,3 GLN
39 100,1 GABA
13 39,3 SAR
40 101,2 KYN
14 45,6 AAA
41 102,7 HIS
15 48,9 PRO
42 104 1MHIS
16 50,8 GLY
43 106,2 3MHIS
17 53,7 ALA
44 108,9 TRP
18 56,3 CIT
45 112,7 CAR
19 59,5 ABU
46 113,7 HCAR
20 65,9 VAL
47 117,1 ANS
21 67,6 SAC
48 119,6 AEC
22 68 CYS2
49 120,3 HYL
23 68,6 ASA
50 121,8 ORN
24 69,2 PIP
51 123,6 LYS
25 69,7 CYSTA
52 126,6 Ammonia
26 70,2 MET
53 128,9 ETH
54 135,1 ARG
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
Seite 3 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Seite 4 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Clinical
Applications
Seite 5 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Physiological (125x3/iVD)
Equipment .................... : ARACUS
Column ......................... : Lithium system, 125mm BL x 3.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent A to Eluent F
Runtime ........................ : 103 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 180 µl/min
Column Temperature ... : 35 °C to 70 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 90 µl/min
Reactor Temperature ... : 115 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Physiological standard Solution
Standard Solution with 100 nmol/ml (Channel A / 570nm)
3.9
95
' P
-Ser
5.3
29
' T
au
6.2
55
' P
EA
7.9
23
' U
rea
18
.00
8'
Asp
20
.63
8'
Hyp
ro
24
.34
6'
Th
r
26
.46
0'
Ser
29
.74
7'
Asn 31
.45
1'
Glu
34
.46
6'
Sar
35
.95
5'
a-A
AA
37
.80
6'
Pro
39
.55
8'
Gly
40
.96
6'
Ala
42
.61
6'
Cit
43
.95
3'
a-A
BA
47
.23
9'
Val
49
.21
0'
(Cy
s)2
50
.73
3'
Cy
sth
a5
1.3
95
' M
et
52
.79
4'
Ile
53
.97
8'
Leu
55
.23
6'
Nle
56
.91
7'
Ty
r
60
.27
2'
Ph
e
66
.88
1'
H-C
yst
ine
69
.97
4'
b-A
la7
0.4
20
' b
-AiB
A
71
.81
9'
g-A
BA
74
.17
4'
His
75
.00
2'
1M
ehis
76
.36
8'
3M
ehis
78
.81
5'
Trp
80
.34
9'
Car
82
.98
1'
Ans
90
.41
6'
Hyly
s
91
.64
2'
Orn
93
.12
5' L
eu
95
.50
0'
NH
4
97
.34
6'
EO
HN
H2
10
1.5
66' A
rg
Seite 6 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Physiological (125x3/iVD)
number retention time
code
number retention time
code [min]
[min]
1 42616 P-Ser
22 52.8 Ile
2 42434 Tau
23 53.9 Leu
3 42406 PEA
24 55.2 Nle
4 42620 Urea
25 56.9 Tyr
5 18.0 Asp
26 60.2 Phe
6 42541 Hypro
27 66.9 H-Cystin
7 42453 Thr
28 69.9 b-Ala
8 42516 Ser
29 70.4 b-Aiba
9 42580 Asn
30 71.8 g-ABA
10 42521 Glu
31 74.1 His
11 34.5 Sar
32 75.0 1Mehis
12 35.9 a-AAA
33 76.4 3Mehis
13 37.8 Pro
34 78.8 Trp
14 39.6 Gly
35 80.3 Car
15 40.9 Ala
36 82.9 Ans
16 42.6 Cit
37 90.4 Hylys
17 43.9 a-ABA
38 91.6 Orn
18 47.2 Val
39 93.1 Lys
19 49.2 (Cys)2
40 95.5 NH4
20 50.7 Cystha
41 97.3 EOHNH2
21 51.4 Met
42 101.6 Arg
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
Seite 7 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Physiological (150x4)
Equipment .................... : ARACUS
Column ......................... : Lithium system, 150 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent A to Eluent F
Runtime ........................ : 128 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 240 µl/min
Column Temperature ... : 35 °C to 74 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 120 µl/min
Reactor Temperature ... : 125 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Physiological standard Solution
Standard Solution with 100 nmol/ml (Channel A / 570nm)
4.1
28
' P
-Ser
6.1
35
' T
au7
.30
9'
PE
A
9.6
91
' U
rea
21
.02
8'
Asp
24
.00
3'
Hyp
ro
28
.17
3'
Th
r
30
.29
2'
Ser
34
.37
7'
Asn
36
.68
4'
Glu
39
.42
0' G
ln
42
.48
7'
Sar
45
.29
7'
a-A
AA
48
.44
6'
Pro
50
.24
1'
Gly
52
.86
5'
Ala
55
.06
0'
Cit
58
.05
3'
a-A
BA
63
.93
8'
Val
66
.46
8'
(Cy
s)2
67
.99
9'
Cy
sth
a6
9.3
40
' M
et
71
.45
0'
Ile
73
.14
5'
Leu
75
.01
0'
Nle
77
.46
0'
Ty
r
82
.66
2'
Ph
e
85
.74
6'
H-C
yst
ine
90
.95
0'
b-A
la
93
.85
4'
b-A
iBA
97
.10
2'
g-A
BA
10
0.4
10' H
is
10
1.7
46' 1
Meh
is
10
4.2
38' 3
Meh
is
10
5.5
99' T
rp
11
1.0
49' C
ar
11
3.9
05' A
ns
11
5.9
27' H
yly
s1
17
.18
1' O
rn
11
8.5
68' L
ys
12
1.7
72' N
H4
12
3.7
54' E
OH
NH
2
12
9.7
96' A
rg
Seite 8 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Physiological (150x4)
number retention time
code
number retention time
code [min]
[min]
1 4.1 P-Ser
23 71.5 Ile
2 6.1 Tau
24 73.1 Leu
3 7.3 PEA
25 73.1 Nle
4 9.7 Urea
26 75.0 Tyr
5 21.0 Asp
27 77.5 Phe
6 24.0 Hypro
28 82.7 H-Cystin
7 28.2 Thr
29 85.7 b-Ala
8 30.3 Ser
30 90.9 b-Aiba
9 34.4 Asn
31 93.9 g-ABA
10 36.7 Glu
32 97.1 His
11 39.4 Gln
34 100.4 1Mehis
12 42.5 Sar
35 101.7 3Mehis
13 45.3 a-AAA
36 104.2 Trp
14 48.4 Pro
37 105.6 Car
15 50.2 Gly
38 111.0 Ans
16 52.9 Ala
39 113.9 Hylys
17 55.1 Cit
40 115.9 Orn
18 58.1 a-ABA
41 117.2 Lys
19 63.9 Val
42 121.8 NH4
20 66.5 (Cys)2
43 123.8 EOHNH2
21 67.9 Cystha
44 129.8 Arg
22 69.3 Met
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
Seite 9 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Diagnostic short (MSUD)
Equipment .................... : ARACUS
Column ......................... : Lithium system, 150 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent B, C and Eluent F
Runtime ........................ : 34 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 240 µl/min
Column Temperature ... : 57 °C to 70 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 120 µl/min
Reactor Temperature ... : 125 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Diagnostic standard solution
Standard Solution with 100 nmol/ml (Channel A / 570nm)
17
.46
3'
Val
19
.54
6'
Met
20
.48
1'
a-Il
e
21
.60
4'
Ile
22
.68
9'
Leu
24
.48
2'
Nle
25
.92
7'
Ty
r
31
.39
3'
Ph
e
Seite 10 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Diagnostic short (MSUD)
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
number retention time
[min] code
1 17.5 Val
2 19.5 Met
3 20.5 a-Ile
4 21.6 Ile
5 22.7 Leu
6 24.5 Nle
7 25.9 Tyr
8 31.4 Phe
Seite 11 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Seite 12 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Food and
Feedstuff
Applications
Seite 13 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Hydrolysate (125x3)
Equipment .................... : ARACUS
Column ......................... : Lithium system, 125 mm BL x 3.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent A to Eluent F
Runtime ........................ : 60 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 220 µl/min
Column Temperature ... : 49 °C to 70 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 110 µl/min
Reactor Temperature ... : 115 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Hydrolysate standard solution
Standard Solution with 100 nmol/ml (Channel A / 570nm)
12
.74
7' A
sp
18
.36
9'
Th
r
19
.58
6'
Sar
20
.86
6'
Glu
24
.70
8'
Pro
26
.13
9'
Gly
27
.43
1'
Ala
28
.44
2' (
Cy
s)2
28
.94
4'
Val
30
.54
2'
Met
32
.14
1'
Ile
32
.91
3'
Leu
34
.49
1'
Ty
r
35
.73
2' P
he
39
.04
8'
His
44
.67
4'
Ly
s
47
.07
9'
NH
4
52
.15
4' A
rg
Channel A
Seite 14 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Hydrolysate (125x3)
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
number retention time
[min] code
1 12.7 Asp
2 18.3 Thr
3 19.6 Ser
4 20.9 Glu
5 24.7 Pro
6 26.1 Gly
7 27.4 Ala
8 28.4 (Cys)2
9 28.9 Val
10 30.5 Met
11 32.1 Ile
12 32.9 Leu
13 34.5 Tyr
14 35.7 Phe
15 39.0 His
16 44.7 Lys
17 47.1 NH4
18 52.2 Arg
Seite 15 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Hydrolysate (125x4)
Equipment .................... : ARACUS
Column ......................... : Lithium system, 125 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent A to Eluent F
Runtime ........................ : 60 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 280 µl/min
Column Temperature ... : 49 °C to 70 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 140 µl/min
Reactor Temperature ... : 125 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Hydrolysate standard solution
Standard Solution with 100 nmol/ml (Channel A / 570nm)
15
.43
7'
Asp
23
.11
4'
Th
r
24
.53
5'
Ser
25
.81
6'
Glu
31
.09
6'
Pro
32
.57
8'
Gly
34
.72
0'
Ala 3
5.4
17
' (C
ys)
2 36
.22
2'
Val
38
.09
7'
Met
40
.09
7'
Ile
41
.06
6'
Leu 4
2.6
68
' T
yr
44
.35
4' P
he
45
.64
2'
His
50
.56
9' L
ys
53
.92
2'
NH
4
59
.55
4'
Arg
Channel A Channel B
Seite 16 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Hydrolysate (125x4)
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
number retention time
[min] code
1 15.4 Asp
2 23.1 Thr
3 24.5 Ser
4 25.8 Glu
5 31.1 Pro
6 32.6 Gly
7 34.7 Ala
8 35.4 (Cys)2
9 36.2 Val
10 38.1 Met
11 40.1 Ile
12 41.1 Leu
13 42.7 Tyr
14 44.4 Phe
15 45.6 His
16 50.6 Lys
17 53.9 NH4
18 59.6 Arg
Seite 17 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Hydrolysate PLUS (TRP)
Equipment .................... : ARACUS
Column ......................... : Lithium system, 125 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent A to Eluent F
Runtime ........................ : 80 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 220 µl/min and 280µl/min
Column Temperature ... : 33 °C to 70 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 120µl/min and 140 µl/min
Reactor Temperature ... : 125 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Hydrolysate standard solution with additional:
L-Asparagine
L-Tryptophan
L-Ornithine
Standard Solution with 100 nmol/ml (Channel A / 570nm)
25
.19
2'
Asp
33
.73
4'
Th
r
36
.17
2'
Ser
40
.60
6'
Asn
41
.67
0'
Glu
47
.17
8'
Pro
48
.72
2'
Gly
50
.15
2'
Ala
50
.95
3'
(Cy
s)2
51
.71
4'
Val
54
.33
4'
Met
56
.19
8' I
le
57
.29
8'
Leu
58
.65
6'
Ty
r
60
.61
2'
Ph
e
61
.77
4'
His
64
.78
7'
Trp
65
.98
1'
Orn
68
.83
2'
Ly
s
72
.86
4'
NH
4
79
.05
5'
Arg
Channel A
Seite 18 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Hydrolysate PLUS (TRP)
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
number retention time
[min] code
1 25.2 Asp
2 33.7 Thr
3 36.2 Ser
4 40.6 Asn
5 41.6 Glu
6 47.2 Pro
7 48.7 Gly
8 50.2 Ala
9 50.9 (Cys)2
10 51.7 Val
11 54.3 Met
12 56.2 Ile
13 57.3 Leu
14 58.7 Tyr
15 60.6 Phe
16 61.8 His
17 64.8 Trp
18 65.9 Orn
19 68.8 Lys
20 72.9 NH4
21 79.1 Arg
Seite 19 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Feedstuff
Equipment .................... : ARACUS
Column ......................... : Natrium system, 125 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Natrium system - Eluent A to Eluent F
Runtime ........................ : 52 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 250 µl/min
Column Temperature ... : 58 °C to 75 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 125 µl/min
Reactor Temperature ... : 115 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Feedstuff standard
Standard Solution with 100 nmol/ml (Channel A / 570nm)
3.8
22
' C
ya
14
.89
1'
Me
tSO
N
16
.58
2'
Asp
18
.24
6'
Th
r
19
.90
5'
Ser
23
.58
7'
Glu
24
.63
7'
Pro
27
.23
3'
Gly
28
.25
2'
Ala
30
.79
0'
(Cy
s)2
32
.01
3'
Val
33
.70
1'
Me
t
35
.85
8'
Ile
36
.80
4'
Leu
38
.85
1'
Ty
r
40
.28
2'
Ph
e
43
.49
5'
His
46
.21
4'
Ly
s
47
.55
2'
NH
4
52
.50
9'
Arg
Chann
Seite 20 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Feedstuff
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
number retention time
[min] code
1 3.8 Cya
2 14.9 MetSON
3 16.6 Asp
4 18.2 Thr
5 19.9 Ser
6 23.6 Glu
7 24.6 Pro
8 27.2 Gly
9 28.3 Ala
10 30.8 (Cys)2
11 32.0 Val
12 33.7 Met
13 35.9 Ile
14 36.8 Leu
15 38.9 Tyr
16 40.3 Phe
17 43.5 His
18 46.2 Lys
19 47.6 NH4
20 52.5 Arg
Seite 21 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
24 Amino Acids Feedstuff-short
Equipment .................... : ARACUS
Column ......................... : Natrium system, 125 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Natrium system - Eluent A to Eluent F
Runtime ........................ : 26.5 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 450 µl/min
Column Temperature ... : 50 °C and 75 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 215 µl/min
Reactor Temperature ... : 130 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Feedstuff standard solution with additional:
Diaminopimelic acid
γ-Amino-n-butyric Acid
L-Ornithine and
L-Tryptophan
Standard Solution with 100 nmol/ml (Channel A / 570nm)
1.9
59
' C
ya
7.8
51
' M
etS
ON
8.4
47
' A
sp
8.9
96
' T
hr
9.7
58
' S
er
11
.49
5'
Glu
11
.74
5'
Pro
13
.14
3'
Gly
13
.57
2'
Ala
14
.29
4'
(Cy
s)2 1
5.5
20
' V
al
16
.11
7'
DA
P
16
.47
4'
Met
17
.55
3'
Ile
18
.07
6'
Leu
19
.14
2'
Ty
r
20
.53
8'
Ph
e
21
.66
1'
g-A
BA
22
.84
8'
His
23
.41
2'
Orn
23
.88
6'
Ly
s
24
.41
7'
NH
4
25
.67
6'
Trp
26
.44
5'
Ph
e
Channel A
40
80
120
160
200
240
280
320
360
400
440
480
mV
3 6 9 12 15 18 21 24 27 min
Seite 22 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
24 Amino Acids Feedstuff-short
Concentration: 100 nmol amino acid / ml ((Cys) 2: 50 nmol / ml)
number retention time
[min] code
1 1.6 Cya
2 7.9 MetSON
3 8.4 Asp
4 9.0 Thr
5 9.8 Ser
6 11.5 Glu
7 12.2 Pro
8 13.1 Gly
9 13.6 Ala
10 14.3 (Cys)2
11 15.5 Val
12 16.1 DAP
13 16.5 Met
14 17.6 Ile
15 18.1 Leu
16 19.1 Tyr
17 20.5 Phe
18 21.7 g-ABA
19 22.8 His
20 23.4 Orn
21 23.9 Lys
22 24.4 NH4
23 25.7 Trp
24 26.4 Arg
Seite 23 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Seite 24 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Biogene
amine and
others
Seite 25 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Biogenic Amine
Equipment .................... : ARACUS
Column ......................... : Potassium system, 118 mm BL x 2.0 mm ID
Inline Filter, 40 mm x 2.0 mm ID
Eluents .......................... : Potassium system - Eluent A to Eluent D & Eluent F
Runtime ........................ : 40 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 220 µl/min
Column Temperature ... : 75 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 110 µl/min
Reactor Temperature ... : 130 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Biogenic Amine Standard solution
Standard Solution with 100 nmol/ml (Channel A / 570nm)
8.5
88
' D
op
amin
e
10
.89
8'
His
tam
ine
15
.51
1'
Pu
tre
scin
e
19
.46
4'
Cad
ave
rin
e
21
.59
6'
Sp
erm
idin
e
27
.32
6'
Sp
erm
ine
33
.11
5'
Ag
ma
tin
e
35
.12
4'
Ty
ram
ine
Seite 26 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Biogenic Amine
number retention time
[min] code
1 8.6 Dopamine
2 10.9 Histamine
3 15.5 Putrescine
4 19.5 Cadaverine
5 21.6 Spermidine
6 27.3 Spermine
7 33.1 Agmatine
8 35.1 Tyramine
Seite 27 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Glyphosate and AMPA
Equipment .................... : ARACUS
Column ......................... : Lithium system, 118 mm x 3.0 mm ID and
Ammonium Absorber, 40 mm x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent A and Eluent F
Gradient ........................ : Program in Steps
Eluent Flow ................... : 180 µl/min
Column Temperature ... : 60 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 90 µl/min
Reactor Temperature ... : 130 °C
Detection ...................... : Visible – 570 nm
Sample .......................... : Glyphosate
AMPA (Aminomethylphosphonic acid)
Standard Solution with 100 nmol/ml
Gly
ph
osa
te
AM
PA
0
20
40
60
80
100
120
140
160
180
200
mV
0.9 1.8 2.7 3.6 4.5 5.4 6.3 7.2 8.1 9 9.9 min
Seite 28 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Glyphosate and AMPA
Standard Solutions with 100 nmol/ml, 25 nmol/ml and 5 nmol/ml GA_5
GA_2 5
GA_1 00
20
40
60
80
10 0
12 0
14 0
16 0
18 0
20 0
mV
0.9 1.8 2.7 3.6 4.5 5.4 6.3 7.2 8.1 9 9.9 min
Gly
ph
osa
te
AM
PA
Standard Solution of Amino Acids with 100 nmol/ml
P-S
er
Tau
PE
A
Ure
a
30
60
90
120
150
180
210
240
270
300
mV
0.9 1.8 2.7 3.6 4.5 5.4 6.3 7.2 8.1 9 9.9 min
Seite 29 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Stress factors plants
Equipment .................... : ARACUS
Column ......................... : Lithium system, 125 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm BL x 2.0 mm ID
Eluents .......................... : Lithium system - Eluent B to Eluent F
Runtime ........................ : 23 min
Gradient ........................ : Program in Steps
Eluent Flow ................... : 450 µL/min
Column Temperature ... : 50 °C and 70 °C
Injection Volume .......... : 20 µl
Reagent ........................ : Ninhydrin (Post-Column Derivatization)
Reagent Flow ............... : 225 µL/min
Reactor Temperature ... : 125 °C
Detection ...................... : Visible – 570 nm (Channel A) / 440 nm (Channel B)
Sample .......................... : Physiological Standard
Equipment .................... : ARACUS
Column ......................... : Lithium system, 125 mm BL x 4.0 mm ID
Ammonium Absorber, 40 mm BL x 2.0 mm ID
Seite 30 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
A P P L I C A T I O N N O T E
Stress factors plants
Standard Solution with 100 nmol/ml (Channel A / 570nm an Channel B /440nm)
number retention time
[min] amino acid code
1 11.0 L- Proline Pro
2 22.0 L-Arginine Arg
10
.98
3'
Pro
22
.02
1'
Arg
33
.99
7'
Pro
Channel A Channel B
Seite 31 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Definitions of abbreviations
amino acid code
S-Sulfocysteine SULFO
Taurine TAU
o-Phosphoethanolamine PEA
Urea Urea
D-Glucosaminic Acid GLUA
L-Aspartic Acid ASP
Hydroxy-L-Proline HYP
L- Threonine THR
L-Serine SER
L- Asparagine ASN
L-Glutamic Acid GLU
L-Glutamine GLN
L- Sarcosine SAR
L- α-Aminoadipic Acid AAA
L- Proline PRO
Glycine GLY
L-Alanine ALA
L-Citrulline CIT
L-α-Amino-n-butyric Acid ABU
L- Valine VAL
L-Saccharopine SAC
L-Cystine CYS2
Argininosuccinic acid ASA
L-Pipecolic Acid PIP
Cystathionine CYSTA
L-Methionine MET
L-Allo-Isoleucine AILE
L-Isoleucine ILE
L-Leucine LEU
Norleucine(internal standard) NORLE
L-Tyrosine TYR
L-Phenylalanine PHE
H-Cystine HCY2
ß-Alanine BALA
Seite 32 ___________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________
membraPure GmbH · Wolfgang-Küntscher-Straße 14 · D-16761 Hennigsdorf
Tel.: +49 (0)3302 201 20-0 · Email: [email protected]
Succinic anhydride ANHYD ASA
ß-Aminoisobutyric Acid BAIB
Amino-Levulinic Acid ALEVA
γ-Amino-n-butyric Acid GABA
Kynurenine KYN
L-Histidine HIS
1-Methyl-L-Histidine 1MHIS
3-Methyl-L-Histidine 3MHIS
L-Tryptophan TRP
L-Carnosine CAR
Homocarnosine HCAR
L-Anserine ANS
S-Aminoethyl-L-cysteine AEC
Hydroxylysine HYL
L-Ornithine ORN
L-Lysine LYS
Ammonium Chloride Ammonia
Ethanolamine ETH
L-Arginine ARG