Agilent 6495 Triple Quadrupole LC/MS System
Experience A New Level of
Confidence
Agilent Technologies
7/31/2014
Agilent 6495 QQQ LC/MS System
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Overview of Topics
Markets and Applications
Technology, performance, and differentiation
• Technology innovations
• New sensitivity performance standard: Instrument Detection Limit (IDL)
• Quantitation performance
• New software capabilities
Key Applications
• Food safety – Pesticides in food matrices
• Food safety – Estrogens in milk
• Environmental – Water analysis
• Peptide quantitation
Summary: Product Values
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Agilent 6495 QQQ LC/MS System
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6495 Markets and Applications
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Agilent 6495 QQQ LC/MS System
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• Biomarkers
• Peptide Quan
• High sensitivity
• Complex matrices
Protein Quant
• Vitamin D
• Endogenous steroids
Clinical Research
• Pesticides
• Mycotoxins
• Antibiotics
• Sample dilution
Food Safety
• Water
• Hormones
• Contaminants
• Direct injections
Environmental
• Research
• Chemical
• High sensitivity
Academia
• Drug discovery
• ADME/PK
• Bioanalysis
• Microdosing
Pharma
• Kits & Methods
• tMRM
Veterinary Drugs
• Drugs of abuse
• Designer drugs
• Oral fluids
Forensic Toxicology
Proven iFunnel Technology
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Agilent 6495 QQQ LC/MS System
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Agilent Jet Stream Hexabore Capillary Dual Ion Funnel
• Thermal gradient focusing
• Efficient desolvation
• Creates an ion rich zone
• Six capillary inlets
• Samples x10 times more
ion rich gas
• Removes the gas but
captures the ions
• Removes neutral noise
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6495 QQQ Technologies Continued Development
1 2
3
• New Detector with High Energy Conversion Dynode
• Improved NEG ion detection with low noise 3
• New Tapered Hexapole Collision Cell
• Effective ion collection and transmission 2
• New Enhanced Q1 Ion Optics
• Improved ion transmission 1
Proven iFunnel Technology • Agilent Jet Stream
• Hexabore Capillary
• Dual Ion Funnel
• Increased ion generation
• Enhanced ion sampling
Quantitative Applications
• Enhanced peak area response
• Improved peak area %RSD
• More sensitive and precise
• Lower Limits of Detection
(IDL)and Quantitation (LLOQ)
• More reliable and robust
New Enhanced Q1 Ion Optics Improved transmission of ions and system robustness
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1. Reduced Octopole length 1
1. New prefilter 2
1
2
• New optimized MS 1 prefilter geometry for improved precursor ion transmission
– Improved peak area response and peak area %RSD, more sensitive and precise
• New optical lens elements for reduced the probability of contamination and easier autotune
– More reliable and robust performance
New Tapered Hexapole Collision Cell Effective collection and transmission of ions
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• Curved and Tapered Hexapole Assembly
for efficient collection and transmission of
product ions
• Consistent collision cell pressure for
higher quality MS/MS spectra
• Designed for consistent collision energies
across all QQQ platforms
• Capable of higher RF voltage on rods for
higher mass transmission
New Detector with High Energy Conversion Dynode More efficient detection of ions with low noise characteristics
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• Improved ion detection efficiency with High Energy Dynode (HED) voltage up to 20 kV
- Improved peak area response and peak area %RSD in positive & negative ion mode
- Improved sensitivity and precision for a wide m/z range of product ions
• Low noise level at 20 kV
- Improved signal to noise
Gain in Signal with Higher HED Voltage: Anti-EHR2/neu mAB
Peptides
60 – 120% increase in
response to 20kV vs. 10KV Product Ion (m/z)
Resp
onse
Incre
ase
(R
ela
tive
to 1
0K
V) Gain in Signal with Higher HED Voltage:
Anti-HER2/neu mAB Peptides
6495 QQQ - Premium Performance
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Improved sensitivity and lower LLOQs – Average 3x in S/N specifications and applications
Improved precision and excellent accuracy at the lowest levels – 3x in IDL specifications
Proven 6 orders of linear dynamic range
Proven robustness in complex matrix – Food matrix and biological matrix (plasma)
Improved mass range, fast scan speed and MRM acquisition rate
Improved Sensitivity – Peak Area Response and S/N Reserpine (+) and Chloramphenicol (-), 1 pg on-column
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Agilent 6495 QQQ LC/MS System
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6495 QQQ
Avg. Area = 14,519
Avg. S/N = 393,000:1
6490 QQQ
Avg. Area = 5,581
Avg. S/N = 157,000:1
6495 QQQ
Avg. Area = 8,808
Avg. S/N = 315,000:1
6490 QQQ
Avg. Area = 3,318
Avg. S/N = 125,000:1
4 x10
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
0.55
0.6
0.65
0.7
0.75
0.8
0.85
0.9
0.95
1
Counts vs. Acquisition Time (min)
0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8
3 x10
0
0.25
0.5
0.75
1
1.25
1.5
1.75
2
2.25
2.5
2.75
3
3.25
3.5
3.75
4
4.25
4.5
4.75
5
5.25
5.5
Counts vs. Acquisition Time (min)
0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6
1 pg reserpine (+) (n=10) 1 pg chloramphenicol (-) (n=10)
2.6 x higher area response
2.5x higher S/N
2.6 x higher area response
2.5x higher S/N
• The 6495 QQQ LC/MS system shows improved peak area response and S/N, in both positive
and negative ion modes compared to previous designs
Many Ways to Manipulate S/N
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Variation in S/N measurements makes direct assessment difficult
Increase signal
• Increase the gain
• Narrow chromatographic peak width
• Increase scan averaging
Lower noise
• Select noise region
• Narrow the width of noise region
• Adjust baseline
• Apply peak smoothing & noise filtering
• Vary noise calculation algorithms: Peak-to-Peak, RMS, and ...
a. Peak-to-peak noise
b. RMS noise
c. Smoothing
d. Baseline noise filtering
Instrument Detection Limit (IDL) is Defined by Statistics
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IDLLCMS = t x SD = t x (%RSD / 100) x amount measured
Based on a well-established statistical formula, follows regulatory guidelines
• The minimum amount of analyte that is detectable and distinguishable from background noise with a confidence level
IDL
• Student “t” value, for
• 99% confidence level
• n – 1 degree of freedom t
• Relative standard deviation / precision of peak area at the amount measured
• From n replicate injections %RSD
• Limited to 2 – 5 x times higher than the Detection Limit (DL)
Amount measured
• Theoretical fitting of %RSD is based on
ion statistics
• %RSD increases at lower injected
amount
%RSD vs. Injected Amount
Why Add IDL Specs for QQQ LC/MS?
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S/N
Many factors impact S/N: - Lower noise - Increase signal
Significant variation in S/N measurements
A relatively high level is used for S/N measurement
Fails to estimate the true limits of detection and quantitation (LLOQ)
Not a good metric of sensitivity performance
IDL (%RSD)
Access sensitivity performance from area %RSD (precision) of replicate injections
Based on a well established statistical formula – follows IUPAC / EPA guidelines
An analytical low level is used for IDL measurement – determined using calibration curve
Accurate assessment of the true limits of detection and quantitation (LLOQ)
A better and more rigorous sensitivity performance metric
Improved Sensitivity and Precision – Peak Area %RSD Reserpine (+) and Chloramphenicol (-) at low levels, 5 fg on-column
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6495 QQQ
Avg. Area = 157
Area %RSD = 6.1
6490 QQQ
Avg. Area = 47
Area %RSD = 10.4
6495 QQQ
Avg. Area = 121
Area %RSD = 2.7
6490 QQQ
Avg. Area = 40
Area %RSD = 5.6
5 fg reserpine (+) (n=10) 5 fg chloramphenicol (-) (n=10)
3 x higher area response
Lower area %RSD
3 x higher area response
Lower area %RSD
• The improved sensitivity of the 6495 QQQ LC/MS system results in enhanced peak area
response and improved area precision (%RSD), especially at the low levels
• This ultimately leads to lower instrument detection limits (IDLs) compared to previous designs
2 x10
0.35
0.4
0.45
0.5
0.55
0.6
0.65
0.7
0.75
0.8
0.85
0.9
0.95
1
1.05
1.1
1.15
1.2
1.25
1.3
1.35
Counts vs. Acquisition Time (min) 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9
2 x10
0.35
0.4
0.45
0.5
0.55
0.6
0.65
0.7
0.75
0.8
0.85
0.9
0.95
1
1.05
1.1
1.15 1
Counts vs. Acquisition Time (min) 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6
Improved Sensitivity and Precision – 6495 QQQ IDL
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1 fg of reserpine used to measure IDL (+) 1 fg of chloramphenicol used to measure IDL (-)
IDL = t x (%RSD / 100) x Amount measured
= 2.821 x (7.2 / 100) x 1 fg
= 0.20 fg
IDL = t x (%RSD / 100) x Amount measured
= 2.821 x (9.7 / 100) x 1 fg
= 0.27 fg
6495 QQQ IDL Amount measured Replicates Area %RSD t (99%) IDL
Reserpine (+) 1 fg n = 10 7.2 2.821 0.20 fg
Chloramphenicol (-) 1 fg n = 10 9.7 2.821 0.27 fg
5 fg
5 fg
1.02 fg
1.25 fg
7.2
8.9
Improved precision (%RSD) and Instrument Detection Limit (IDL) achieved on the 6495 vs. the 6490
IDL for the Agilent 6495 QQQ LC/MS System
1 x10
5.4
5.6
5.8
6
6.2
6.4
6.6
6.8
7
7.2
7.4
7.6
7.8
3.6 3.7 3.8 3.9 4 4.1 4.2 4.3
… 1 x10
5.4
5.6
5.8
6
6.2
6.4
6.6
6.8
7
7.2
7.4
7.6
7.8
Acquisition Time (min) 1.45 1.55 1.65 1.75 1.85 1.95 2.05
Acquisition Time (min)
6495 IDL Levels: Accurate Assessment of the LLOQs Reserpine (+) and Chloramphenicol (-)
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Agilent 6495 QQQ LC/MS System
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6495 QQQ - Reserpine (+) IDL 6495 QQQ – Chloramphenicol (-) IDL
0.20 fg 0.27 fg
6495 QQQ: Reserpine LLOQ = 0.2 fg 6495 QQQ: Chloramphenicol LLOQ = 0.3 fg
Blank Blank
Area % RSD = 15.8
n = 10 injections
%Accuracy = 118
Area % RSD = 18.8
n = 10 injections
%Accuracy = 108
• Improved / Lower IDL levels and LLOQ levels are achieved on the new 6495 vs. the 6490
1 x10
5.3
5.4
5.5
5.6
5.7
5.8
5.9
6
3.6 3.7 3.8 3.9 4 4.1 4.2 4.3
1 x10
5.4
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
1.45 1.55 1.65 1.75 1.85 1.95 2.05
1.02 fg 1.25 fg
6490 QQQ
Chloramphenicol LLOQ = 1.0 fg
6490 QQQ
Reserpine LLOQ = 1.0 fg
6495 QQQ LC/MS Performance Specifications
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Bid Specifications Agilent
6490
Agilent
6495
Sensitivity, S/N
Positive, 1 pg Reserpine on-column 50,000:1 150,000:1
Sensitivity, S/N
Negative, 1 pg chloramphenicol on-column 50,000:1 150,000:1
Sensitivity, Instrument Detection Limit (IDL)
Positive, Reserpine 2.5 fg
Measured at 5 fg on-column
0.75 fg Measured at 1 fg on-column
Sensitivity, Instrument Detection Limit (IDL)
Negative, chloramphenicol 2.5 fg
Measured at 5 fg on-column
0.75 fg Measured at 1 fg on-column
Linear dynamic range 106 106
Mass range 5 – 1,400 Da 5 – 2,250 Da
Maximum scan speed 12,500 Da/sec 15,000 Da/sec
Maximum MRM rate 250
MRMs/sec
500
MRMs/sec
Minimum MRM dwell time 1 ms 1 ms
Polarity switching 30 ms 30 ms
• Improved sensitivity specs: - S/N - IDL
• Improved mass range
• Improved full scan and MRM speed
Verapamil (+) Sensitivity in Solvent Attogram / Zeptomole Lower Limit of Quantitation
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• Unparalleled sensitivity (LLOQ of 40 ag on-column) - better than the 6490 (100 ag)
• Excellent precision (peak area %RSD) and accuracy are observed at the lowest levels
6495 QQQ: Verapamil LLOQ = 40 ag (80 zeptomoles) on-column
Blank
Area %RSD = 15.1
n = 10 injections
%Accuracy = 116
Verapamil 100 ag on-column
Improved LLOQ, 6495 vs. 6490
6495 QQQ
Area %RSD = 11.1
6490 QQQ
Area %RSD = 19.5
Verapamil IDL – Measured at the LLOQ Level
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Amount measured Replicates %RSD t (99%) IDL
40 ag (LLOQ) n = 10 15.1 2.182 17 ag
IDL = t x (%RSD/100) x Amount = 2.182 x (15.1/100) x 1.0 fg = 17 ag
Inj # Peak Area
1 16
2 19
3 19
4 20
5 19
6 13
7 18
8 21
9 16
10 14
%RSD 15.1
• Excellent peak area precision (%RSD) are observed at the lowest level (LLOQ)
1 x10
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
3.827 16
1 x10
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
7 3.824 19
1 x10
5.4
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9 3.824
19
Acquisition Time (min)
3.65 3.7 3.75 3.8 3.85 3.9 3.95 4
1 x10
5.6
5.8
6
6.2
6.4
6.6
6.8
7
7.2 3.820 20
1 x10
5.6
5.8
6
6.2
6.4
6.6
6.8
7
7.2 3.827 19
1 x10
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
7
3.820
13
3.65 3.7 3.75 3.8 3.85 3.9 3.95 4
1 x10
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8 3.824
18
1 x10
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
7 3.824 21
1 x10
5.6
5.8
6
6.2
6.4
6.6
6.8
7 3.824
16
3.65 3.7 3.75 3.8 3.85 3.9 3.95
Acquisition Time (min) Acquisition Time (min)
1 x10
5.5
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9 3.824 14
3.6 3.65 3.7 3.75 3.8 3.85 3.9 3.95 4
Acquisition Time (min)
Inj 1
Inj 2
Inj 3
Inj 4
Inj 5
Inj 6
Inj 7
Inj 8
Inj 9
Inj 10
Bentazon (-) Sensitivity in Solvent Sub-femtogram Limits of Detection and Quantitation
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LLOQ = 0.5 pg/mL = 1.0 fg on-column in Solvent LOD = 0.2 pg/mL = 0.4 fg on-column in Solvent
• Unparalleled sensitivity (LLOQ 0.5 pg/mL and 1.0 fg on-column) – better than the 6490
• Excellent precision (peak area %RSD) and accuracy are observed at the lowest levels
Blank Blank
Injection volume = 2 µL
Area %RSD = 17.6
n = 7 injections
%Accuracy = 91.8
Avg. S/N = 5.9
Noise = Peak to Peak
Injection volume = 2 µL
Area %RSD = 12.4
n = 7 injections
%Accuracy = 85.1
Avg. S/N = 21.9
Noise = Peak to Peak
Bentazon IDL – Measured at the LLOQ Level
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Amount measured Replicates %RSD t (99%) IDL
0.5 pg/mL, 1.0 fg (LLOQ) n = 7 12.4 3.143 0.20 pg/mL, 0.39 fg
IDL = t x (%RSD/100) x Amount = 3.143 x (12.4/100) x 1.0 fg = 0.39 fg
Inj # Peak Area S/N (Peak to peak)
1 46 33.5
2 38 12
3 51 33.1
4 39 17.1
5 40 30.7
6 41 13.2
7 36 12.9
%RSD 12.4 Ave 21.9
Inj 1
Inj 2
Inj 3
Inj 4
Inj 5
Inj 6
Inj 7
• Excellent peak area precision (%RSD) are observed at the lowest level (LLOQ)
Injection volume = 2 µL
6 Orders of Linear Dynamic Range, Peptide Quantitation
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• Excellent assay accuracy and precision are
achieved at all levels including the LLOQ level
LVNEVTEFAK, 5 amol – 5 pmol, In trypsin digest
6 Orders of Linear Dynamic Range
R2 = 0.998
Levels %RSD
(n = 10)
%
Accuracy
RT %RSD
(n = 100)
5 amol 14.0 109.8
0.12
7.5 amol 16.0 108.7
15 amol 9.4 105.0
30 amol 9.0 87.1
300 fmol 1.6 85.2
3 fmol 1.2 81.4
30 fmol 0.6 86.4
300 fmol 0.7 87.4
3 pmol 2.1 105.6
5 pmol 10. 97.5
Zoom-in 5 – 30 amol
Proven System Robustness in Complex Food Matrix: Multi-Residue Pesticide Analysis in Black Tea
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• Pesticide standards analyzed before, in the middle, and after matrix injections
• No signal decrease was observed after 3 days of operation running with heavy food
matrix (black tea)
• %RSD < 6 (n = 6 data files) – Average results of > 250 pesticides analyzed
Overlaid six data files / MRM chromatograms • QC injections – Day 1
• QC injections – Day 2
• QC injections – Day 3
Unrivaled System Robustness in Biological Matrix: Protein Quantitation in Plasma
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• Selected peptides from 42 peptides in the QC sample – normalized to Day 1 response
• Peptide QC samples analyzed daily after every ~25 plasma digest injections
• No significant signal degradation observed after 853 injections of 40 µg plasma digest
per injection and 3.5 weeks of continuous operation
• Response %RSD: 6 - 15
0%
20%
40%
60%
80%
100%
120%
140%
01/06/14 01/11/14 01/16/14 01/21/14 01/26/14 01/31/14 02/05/14
Apolipoprotein E
L-selectin
Plasminogen
Albumin_serum
Kininogen
Transthyretin
Hemopexin
Day of Analysis
Resp
on
se
Ultra-Fast MRM Acquisition – 1ms Dwell Time
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Calibration Curve at 1ms dwell
1 – 1000 ng/mL
123 Pesticides, 1 ng/mL
• Fast MRM allows enough data
points across the peak for precise
and accurate multi-compound
quantitation with UHPLC speed
Carbosulfan, 18 data points
1,000 ng/mL
%Accuracy = 100.4
100 ng/mL
%RSD = 4.2
%Accuracy = 99.5 1 ng/mL
%RSD = 4.4
%Accuracy = 108.3
10 ng/mL
%RSD = 6.2
%Accuracy = 92.1
• Precision %RSD < 20
• %Accuracy 80 - 120
Improved Mass Range
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• Improved peptide quantitation can be achieved with the 6495
- Extended mass range up to 2,250 m/z
- Increased detection efficiency (HED voltage up to 20kV)
6495 QQQ MRM Spectrum for Glycopeptide:
EEQYN[+1606.6]STYR (G1F)
m/z 2227.88
6495 QQQ Key Applications
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Food Safety - Pesticides
Analysis of >250 trace level pesticides in food matrices (e.g. black tea) with extensive sample dilutions
Food Safety - Estrogens
Determination of ultra-trace level estrogens in milk product
Environmental - Water Analysis
Quantitation of ultra-trace level hormones (EDCs) in drinking water using direct injection
Peptide Quantitation
Quantitation of synthetic peptide at sub-attomole level using nanoflow and standard flow chromatography
Peptide Quantitation
Quantitative analysis of microcystins (cyclic nonribosomal peptides) in liver extract samples using stand flow chromatography
High Sensitivity Allows Extensive Sample Dilution
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1:20 dilution of > 250 pesticides spiked into black tea at 10 µg/kg (MRL), 0.1 ng/mL
1:100 dilution of pesticides in black tea at MRL, 20 pg/mL
• Multi-residue pesticide analysis in food products – most demanding food safety applications
• Improved sensitivity and precision of the 6495 allows accurate quantitation of pesticides
< Maximum Residue Limits (MRLs) imposed by EU, even with extensive sample dilution
• Sample dilution leads to less matrix effects and improved method robustness
Injection volume = 2 µL
Ultimate Sensitivity for Pesticides in Negative Mode Sub-pg/mL LLOQs and Sub-fg IDLs
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Pesticides
Fipronil
Bentazon
Teflubenzuron
Hexaflumuron
Flubendiamide
Fluazinam
Black Tea Matrix
LOD
(pg/mL)
LLOQ
(pg/mL)
IDL
pg/mL fg
1 1 0.39 0.78
0.5 1 0.36 0.72
5 5 2.13 4.25
5 5 1.09 3.19
1 5 0.54 1.08
5 5 2.13 4.25
Neat Solvent
LOD
(pg/mL)
LLOQ
(pg/mL)
IDL
pg/mL fg
0.2 0.5 0.14 0.27
0.2 0.5 0.20 0.39
0.5 1 0.37 0.73
0.5 1 0.38 0.75
0.5 1 0.27 0.54
0.5 1 1.10 2.20
Fipronil 1 pg/mL
Bentazon 0.5 pg/mL
Fluazinam 5 pg/mL
Flubendiamide 1 pg/mL
Hexaflumuron 5 pg/mL
Teflubenzuron 5 pg/mL
LOD Levels in Black Tea Matrix
Injection volume = 2 µL
• LLOQs < 20 pg/mL (40 fg on-column) is required to achieve MRL with 1:100 dilution
Bentazon Sensitivity in Black Tea Low femtogram Limits of Detection and Quantitation
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LLOQ = 1 pg/mL = 2.0 fg in 20x Diluted Black Tea LOD = 0.5 pg/mL = 1.0 fg in 20x Diluted Black Tea
• Unparalleled sensitivity in black tea matrix - LLOQ 1.0 pg/mL and 2.0 fg on-column
• Excellent precision (peak area %RSD) and accuracy are observed at the low levels in
black tea matrix, e.g. the LLOQ level.
Injection volume = 2 µL
Area %RSD = 16.9
n = 7 injections
Avg. S/N = 7.4
Noise = Peak to Peak
95.79
Injection volume = 2 µL
Area %RSD = 11.4
n = 7 injections
%Accuracy = 109.8
Avg. S/N = 16.1
Noise = Peak to Peak
Bentazon: Superb Precision & IDL in Black Tea
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Amount measured Replicates %RSD t (99%) IDL
1 pg/mL, 2.0 fg (LLOQ) n = 7 injections 11.4 3.143 0.36 pg/mL, 0.72 fg
IDL = t x (%RSD/100) x Amount = 3.143 x (11.4/100) x 2.0 fg = 0.72 fg
Inj # Peak
Area
S/N (Peak to peak)
1 82 20.9
2 96 13.2
3 70 12.7
4 86 13.4
5 92 10.3
6 72 12.7
7 83 29.4
%RSD 11.4 Ave 16.1
Injection volume = 2 µL
• Excellent peak area precision (%RSD) are observed at the lowest level in black tea matrix
1 pg/mL
1 pg/mL
1 pg/mL
1 pg/mL
1 pg/mL
1 pg/mL
1 pg/mL
Five Orders of Linear Dynamic Range
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Zoom-in 1 – 100 pg/mL
• Wide (5 orders) linear dynamic range, excellent assay accuracy and precision at all levels
Levels %RSD (n = 7) %Accuracy
1 pg/mL 10.41 82.4
5 pg/mL 5.84 87.2
10 pg/mL 6.41 82.6
50 pg/mL 6.10 97.4
100 pg/mL 3.86 100.8
500 pg/mL 1.73 108.8
1 ng/mL 2.93 112.5
5 ng/mL 2.25 119.7
10 ng/mL 2.54 116.8
50 ng/mL 2.62 102.3
100 ng/mL 2.51 95.8
Fipronil (-) in 1:20 diluted black tea
1 pg/mL – 100 ng/mL (2 fg - 200 pg)
5 Orders
R2 = 0.995
Spinosyn A (+) in 1:20 diluted black tea
1 pg/mL – 100 ng/mL (2 fg - 200 pg)
5 Orders
R2 = 0.997 Log – Log scale
Levels %RSD (n = 7) %Accuracy
1 pg/mL 15.0 110.7
5 pg/mL 10.9 85.7
10 pg/mL 6.8 97.2
50 pg/mL 3.6 86.0
100 pg/mL 5.4 100.5
500 pg/mL 7.3 96.3
1 ng/mL 6.5 105.0
5 ng/mL 2.1 105.8
10 ng/mL 3.6 111.3
100 ng/mL 1.2 96.2
Signal Improvements for High Relevance Pesticides
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6495 6490 6495 6490
Methidathion
Area Gain 3.7x
Methomyl
Area Gain 3.5x
Buprofezine
Area Gain 4.2x
Monocrotophos
Area Gain 2.9x
Thiabendazole
Area Gain 3.2x
Spiromesifen
Area Gain 2.1x
6495 6490 6495 6490
6495 6490 6495 6490
• Improved sensitivity (peak area gain of 3x) are observed on the new 6495 vs. the 6490
Improved Precision & Lower LLOQs in Black Tea
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• Improved precision (%RSD) are observed, particularly at the lowest levels (LLOQs)
• The enhanced peak area response and improved precision (%RSD) means pesticides
achieve lower LLOQs ( MRL level) using the new 6495.
Comparison of Area %RSD for 50 Pesticides at MRL # of pesticides detected at MRL
• 67% of pesticides were easily detected
with a %RSD < 20 with 1:100 dilution
• This means LLOQs < 20 pg/mL (ppt)
• %RSD < 20 meets SANCO guidelines
6495
Dilution allows More Efficient Ionization of Pesticides
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1 : 5 1 : 10 1 : 20 1 : 100 1 : 50
1 : 5 1 : 10 1 : 20 1 : 100 1 : 50
Alanycarb, 10 µg/kg in Black Tea
No
Dilution
Oxamyl, 10 µg/kg in Black Tea
No
Dilution
Recovery (%) in Black Tea with Different Dilutions
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Pesticides No dilution
(n = 5)
Dilution 1:5
(n = 5)
Dilution 1:10
(n = 5)
Dilution 1:20
(n = 5)
Dilution 1:50
(n = 5)
Dilution 1:100
(n = 5)
Acetamiprid 29.4 ± 0.8 57.3 ± 1.4 67.5 ± 3.7 79.9 ± 2.9 91.8 ± 5.2 109.5 ± 3.4
Alanycarb 10.4 ± 1.3 73.9 ± 2.2 81.5 ± 14.3 85.7 ± 11.1 87.6 ± 4.7 121.7 ± 10.8
Aldicarb 36.9 ± 1.0 69.9 ± 1.4 78.0 ± 3.5 91.0 ± 4.2 95.2 ± 8.8 104.9 ± 14.1
Carbaryl 56.9 ± 1.8 80.1 ± 3.8 80.8 ± 4.1 96.1 ± 7.2 102.6 ± 6.6 116.4 ± 9.6
Dimethoate 33.9 ± 1.7 68.6 ± 2.4 84.1 ± 5.4 89.0 ± 7.9 88.2 ± 8.8 84.7 ± 7.5
Diuron 79.7 ± 4.0 90.4 ± 7.0 91.7 ± 4.9 94.9 ± 7.2 89.2 ± 7.3 100.9 ± 13.5
Flufenoxuron 95.4 ± 1.1 88.8 ± 1.6 89.4 ± 3.8 93.3 ± 5.8 100.0 ± 6.1 119.2 ± 13.9
Monocrotophos 4.6 ± 0.3 13.9 ± 0.3 21.8 ± 0.8 33.8 ± 1.1 58.5 ± 2.0 95.1 ± 5.7
Oxamyl 20.8 ± 0.7 52.6 ± 1.9 65.0 ± 2.0 79.7 ± 3.0 91.2 ± 4.6 110.6 ± 5.2
Thiamethoxam 40.0 ± 1.4 45.9 ± 0.9 46.6 ± 3.8 52.2 ± 1.7 70.9 ± 2.9 97.3 ± 2.0
• Cells shaded in green shows %recovery of 80 – 120, which are in compliance with
SANCO requirements.
• Pesticides show full recovery and no signal suppression with 1:100 dilution
• Neat solvent calibration curve can be used for pesticides quantitation with 1:100 dilution
6495 QQQ Key Applications
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Agilent 6495 QQQ LC/MS System
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Food Safety - Pesticides
Analysis of >250 trace level pesticides in food matrices (e.g. black tea) with extensive sample dilutions
Food Safety - Estrogens
Determination of ultra-trace level estrogens in milk product
Environmental - Water Analysis
Quantitation of ultra-trace level hormones (EDCs) in drinking water using direct injection
Peptide Quantitation
Quantitation of synthetic peptide at sub-attomole level using nanoflow and standard flow chromatography
Peptide Quantitation
Quantitative analysis of microcystins (cyclic nonribosomal peptides) in liver extract samples using stand flow chromatography
Sensitive Quantitation of Estrogens in Cow Milk
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Eight estrogens spiked in cow milk at 0.1 µg/kg, corresponding to 0.1 ng/mL
• Estrogen levels in milk products are regulated by government agencies down to 1 µg/kg
• Highly sensitive and precise quantitation of estrogens was achieved using the new 6495
with a simple QuEChERS procedure without derivatization or enrichment.
• LLOQs for all estrogens (<< 0.1 µg/kg) are far below the regulatory requirement (1 µg/kg)
ID Compounds
1 Estriol (-)
2 17-α-Estradiol (-)
3 17-β-Estradiol (-)
4 Ethynyl estradiol (-)
5 Estrone (-)
6 Diethylstilbestrol (-)
7 Hexestrol (-)
8 Dienestrol (-)
2 3
4
5
6
7,8
1
--- 0.1 ng/mL estrogens in cow milk
--- Milk blank
Injection volume = 5 µL
Highest Sensitivity and Excellent Precision LLOQs and IDLs at pg/mL (fg) level
Compounds LLOQ
(pg/mL)
%RSD
n = 7
IDL
pg/mL fg
Estriol 5 11.3 1.8 8.9
17-α-Estradiol 10 8.4 2.7 13.3
17-β-Estradiol 10 11.4 3.6 18.0
Ethynyl estradiol 10 11.7 3.7 18.5
Estrone 5 9.6 1.5 7.5
Diethylstilbestrol 10 12.8 2.0 10.1
Hexestrol 5 11.1 1.7 8.7
Dienestrol 5 7.6 1.2 6.0
Estriol
5 pg/mL
17-Estradiol
10 pg/mL
17-Estradiol
10 pg/mL
Ethynyl Estradiol
10 pg/mL
Estrone
5 pg/mL
Diethylstilbestrol
10 pg/mL
Hexestrol
5 pg/mL
Dienestrol
5 pg/mL
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Injection volume = 5 µL
Injection volume = 5 µL
Estriol: Superb Precision and IDL at Low-fg Level
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Estriol Amount measured Replicates %RSD t (99%) Estriol IDL
5 pg/mL, 25 fg (LLOQ) n = 7 injections 11.3 3.143 1.8 pg/mL, 8.9 fg
IDL = t x (%RSD/100) x Amount = 3.143 x (10.4/100) x 25 fg = 8.9 fg
Inj # Peak
Area S/N
(Peak to peak)
1 44 11.1
2 54 11.1
3 41 9.8
4 46 9.8
5 50 10.5
6 54 8.6
7 52 9.9
%RSD 11.3 Ave. 10.1
Injection volume = 5 µL
5 pg/mL
5 pg/mL
5 pg/mL
5 pg/mL
5 pg/mL 5 pg/mL
5 pg/mL
• Excellent peak area precision (%RSD) are observed at the lowest levels (LLOQs)
Confident Estrogen Quantitation in Milk
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Levels (µg/kg) %RSD (n = 7) %Accuracy
0.1 5.33 106.0
0.2 2.96 101.2
0.5 2.99 94.6
1 2.75 95.3
2 0.94 100.3
5 1.68 104.1
10 1.64 98.5
Levels (µg/kg) %RSD (n = 7) %Accuracy
0.1 3.86 113.8
0.2 2.61 102.5
0.5 3.68 91.1
1 1.74 92.5
2 1.45 97.9
5 3.64 101.6
10 2.14 100.6
• Excellent linearity (R2 ≥0.998), accuracy and precision are achieved at all levels
17 -Estradiol in milk, 0.1 – 10 µg/kg
0.1 – 10 ng/mL
R2 = 0.999
Estriol in milk, 0.1 – 10 µg/kg
0.1 – 10 ng/mL
R2 = 0.999
Summary of Estrogen Quantitation in Milk Excellent linearity, precision, accuracy and matrix recovery
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Compounds R2 Precision
%RSD, n = 7 %Accuracy
%Recovery
0.2 µg/kg 1 µg/kg 5 µg/kg
Estriol 0.9988 0.94 – 5.33 94.6 – 106.0 92.9 102.8 85.8
17-α-Estradiol 0.9973 1.32 – 5.31 90.6 – 112.0 92.2 99.4 93.4
17-β-Estradiol 0.9985 1.45 – 3.86 91.1 – 113.8 92.9 106.0 96.9
Ethynyl estradiol 0.9984 1.59 – 6.61 90.1 – 114.5 90.2 104.9 94.0
Estrone 0.9987 1.45 – 3.76 92.4 – 110.7 94.0 101.7 96.3
Diethylstilbestrol 0.9985 2.27 – 5.04 94.9 – 104.6 83.5 98.6 91.0
Hexestrol 0.9981 0.88 – 4.58 90.1 – 112.8 86.1 98.4 91.8
Dienestrol 0.9995 1.24 – 4.34 95.6 – 102.7 82.9 92.8 85.3
6495 QQQ Key Applications
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Food Safety - Pesticides
Analysis of >250 trace level pesticides in food matrices (e.g. black tea) with extensive sample dilutions
Food Safety - Estrogens
Determination of ultra-trace level estrogens in milk product
Environmental - Water Analysis
Quantitation of ultra-trace level hormones (EDCs) in drinking water using direct injection
Peptide Quantitation
Quantitation of synthetic peptide at sub-attomole level using nanoflow and standard flow chromatography
Peptide Quantitation
Quantitative analysis of microcystins (cyclic nonribosomal peptides) in liver extract samples using stand flow chromatography
Analysis of Hormones in Drinking Water
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• Endocrine Disrupting Chemical (EDCs) levels in
municipal water supplies are regulated by government
agencies down to ng/L (ppt) level
• The highly sensitive 6495 allows the quantitation of
EDCs at sub ng/L using direct injection
• No need for time consuming offline SPE or enrichment
Estriol 17-β-Estradiol
Testosterone
17-α-Ethynylestradiol Equilin Estrone
Androstenedione
Estrone (-)
Estriol (-)
Testosterone (+)
Androstenedione (+)
Equilin (-)
17--Ethynyl estradiol (-)
17--estradiol (-)
Hormones in drinking water, 5 ng/L (testosterone) to 17.5 ng/L (EE)
Highest Sensitivity and Excellent Precision LLOQs and IDLs at sub-ng/L Level
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Estriol
2 ng/L
Testosterone
0.1 ng/L
Equilin
0.2 ng/L
17-
Ethynylestradiol
1.75 ng/L
17β-Estradiol
0.5 ng/L
Estrone
0.2 ng/L
Androstenedione
0.2 ng/L
Compounds LLOQ
(ng/L)
%RSD
n = 8
IDL
(ng/L)
%
Accuracy
Estriol 1 5.6 0.168 94.8
17β-Estradiol < 0.5 13.5 0.202 102.2
Testosterone 0.1 10.3 0.031 114.5
17α-Ethynylestradiol 1.75 14.8 0.78 98.4
Equilin < 0.2 3.7 0.022 115.1
Estrone < 0.2 7.2 0.043 115.1
Androstenedione < 0.2 4.3 0.026 107.2
17-Estradiol: Superb Precision and Ultra-Low IDL
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17-Estradiol Measured Replicates %RSD t (99%) IDL
0.5 ng/L (LLOQ) n = 8 injections 13.5 2.998 0.202 ng/L
MDL = t x (%RSD/100) x Amount = 2.998 x (13.5/100) x 0.5 ng/L = 0.202 ng/L
Inj # Peak
Area
S/N (Peak to peak)
1 869 9.1
2 925 10.0
3 671 6.7
4 699 6.8
5 950 11.5
6 946 16.0
7 857 8.2
8 762 12.2
%RSD 13.5 10.1
0.5 ng/L
0.5 ng/L
0.5 ng/L
0.5 ng/L
0.5 ng/L 0.5 ng/L
0.5 ng/L
0.5 ng/L
• Excellent peak area precision (%RSD) are observed at the lowest levels (LLOQs)
Confident Quantitation of EDCs in Drinking Water
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• Excellent linearity (R2 ≥0.995), assay accuracy and precision are achieved at all levels
Testosterone (+)
0.1 – 10 ng/L
R2 = 0.996
17-estradiol (-)
0.1 – 10 ng/L
R2 = 0.996
Levels (ng/L) %RSD (n = 8) %Accuracy
0.1 10.3 114.5
0.2 4.89 104.2
0.5 2.71 88.8
1 4.34 92.8
5 4.49 97.4
10 2.09 104.7
Levels (ng/L) %RSD (n = 8) %Accuracy
0.1 13.7 120.3
0.2 13.5 102.2
0.5 3.78 87.5
1 4.14 89.1
5 2.66 98.8
10 0.73 104.2
Summary of Hormone Quantitation in Drinking Water Excellent linearity, precision and accuracy at all levels
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Compounds LLOQ
(ng/L)
IDL
(ng/L) R2 Precision
(%RSD, n = 8) %Accuracy
Estriol 0.4 0.168 0.997 0.81 – 10.6 93.6 – 113.6
17β-Estradiol < 0.5 0.202 0.996 0.73 – 13.7 87.5 – 120.3
Testosterone < 0.1 0.031 0.994 2.09 – 10.3 88.8 – 114.5
17α-Ethynylestradiol 1.75 0.78 0.996 2.50 – 16.5 95.3 – 107.2
Equilin < 0.2 0.022 0.997 3.00 – 5.21 88.9 – 115.1
Estrone < 0.2 0.043 0.996 2.57 – 9.41 91.2 – 115.1
Androstenedione < 0.1 0.026 0.995 1.48 – 9.07 89.2 – 107.2
6495 QQQ Key Applications
7/31/2014
Agilent 6495 QQQ LC/MS System
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Food Safety - Pesticides
Analysis of >250 trace level pesticides in food matrices (e.g. black tea) with extensive sample dilutions
Food Safety - Estrogens
Determination of ultra-trace level estrogens in milk product
Environmental - Water Analysis
Quantitation of ultra-trace level hormones (EDCs) in drinking water using direct injection
Peptide Quantitation
Quantitation of synthetic peptide at sub-attomole level using nanoflow and standard flow chromatography
Peptide Quantitation
Quantitative analysis of microcystins (cyclic nonribosomal peptides) in liver extract samples using stand flow chromatography
Outstanding Sensitivity with Standard Flow LC
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Levels %RSD
(n = 10)
%
Accuracy
RT %RSD
(n = 100)
5 amol 14.0 109.8
0.12
7.5 amol 16.0 108.7
15 amol 9.4 105.0
30 amol 9.0 87.1
300 fmol 1.6 85.2
3 fmol 1.2 81.4
30 fmol 0.6 86.4
300 fmol 0.7 87.4
3 pmol 2.1 105.6
5 pmol 10. 97.5
Zoom-in 5 – 30 amol
LVNEVTEFAK
5 amol – 5 pmol
6 Orders
R2 = 0.998
• Low attomole sensitivity for synthetic peptide spiked in tryptic
digest using the 6495 with standard flow chromatography
• Six orders of linear dynamic range
• Excellent precision and accuracy are observed at all levels
including the LLOQ level
Blank
3 amol
5 amol
Area %RSD = 14.0, n = 10
%Accuracy = 109.8
7.5 amol
LOD
LLOQ
Injection volume = 1 µL
Excellent Precision and Low Attomole Level IDL
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Amount measured Replicates %RSD t (99%) IDL
5.0 amol (LLOQ) n = 10 injections 14.0 2.821 2.0 amol
MDL = t x (%RSD/100) x Amount = 2.821 x (14.0/100) x 5.0 amol = 2.0 amol
Injection # Peak Area
1 30.5
2 28.7
3 30.4
4 33.91
5 35.1
6 31.5
7 34.5
8 20.7
9 26.7
10 26.6
%RSD 14.0
5 amol
5 amol
5 amol
5 amol
5 amol
5 amol
5 amol
5 amol
5 amol
5 amol
• Excellent peak area precision (%RSD) are observed at the lowest levels (LLOQs)
Ultimate Sensitivity with Nanoflow LC
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• Ultimate sub-attomole sensitivity for synthetic peptide spiked
in tryptic digest using the 6495 with nanoflow chromatography
• Wide (over 5 orders) linear dynamic range
• Excellent precision and accuracy are observed at all levels
including the LLOQ level
LVNEVTEFAK
0.5 amol – 0.1 pmol
> 5 Orders
R2 = 0.99996
Levels (amol) %RSD %Accuracy
0.5 4.52 103.2
1 8.50 80.2
10 5.75 84.6
100 4.71 88.9
1000 1.23 85.6
10,000 1.09 97.6
100,000 1.69 100.4
6495 QQQ Key Applications
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Food Safety - Pesticides
Analysis of >250 trace level pesticides in food matrices (e.g. black tea) with extensive sample dilutions
Food Safety - Estrogens
Determination of ultra-trace level estrogens in milk product
Environmental - Water Analysis
Quantitation of ultra-trace level hormones (EDCs) in drinking water using direct injection
Peptide Quantitation
Quantitation of synthetic peptide at sub-attomole level using nanoflow and standard flow chromatography
Peptide Quantitation
Quantitative analysis of microcystins (cyclic nonribosomal peptides) in liver extract samples using stand flow chromatography
Analysis of Microcystins
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• Microcystins are cyanotoxins that may cause serious damage to the liver of mammals
• Microcystins are challenging compounds to analyze due to the low levels in biological matrices
MRM chromatograms of microcystins
Using the 6495 with 1290 UHPLC
Outstanding Sensitivity and Linear Dynamic Range Excellent Peak Area Precision %RSD at the LLOQ Levels (pg/mL)
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Concentration (ng/ml)
0.1 1 10 100
R e
s p
o n s e
s
6 x10
5E-05
0.0001
0.0005
0.001
0.005
0.01
0.05
0.1
0.5
1
R^2 = 0.99495440
Type:Linear, Origin:Ignore, Weight:1/x^2
RR: 2.5-50,000 ng/mL > 4 Orders of Linear Dynamic Range R2 = 0.995
1 x10
4.5
5
5.5
6
6.5
7
7.5
1 x10
4.5
5
5.5
6
6.5
7
7.5 2.297 78.51
Counts vs. Acquisition Time (min)
1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6
Blank
Concentration (ng/ml)
0.05 0.1 0.5 1 5 10 50 100
R e
s p
o n s e
s
4 x10
0.006
0.008
0.01
0.02
0.04
0.06
0.08
0.1
0.2
0.4
0.6
0.8
1
2
4 R^2 = 0.99752628
Type:Linear, Origin:Ignore, Weight:1/x
LY: 50-50,000 ng/mL 3 Orders of Linear Dynamic Range R2 = 0.998
1 x10
4.25
4.5
4.75
5
5.25
5.5
5.75
6
6.25
6.5
1 x10
4.25
4.5
4.75
5
5.25
5.5
5.75
6
6.25
6.5 5.895 56.82
Counts vs. Acquisition Time (min)
5.2 5.4 5.6 5.8 6 6.2
LY, LLOQ 50 pg/mL %RSD = 10.3 n = 3
Blank
• Excellent linearity (R2 ≥ 0.99),
assay accuracy and precision
are achieved for all seven
microcystins and at all levels
RR, LLOQ 2.5 pg/mL %RSD = 2.97 n = 3
Analysis of Microcystins in Liver Extract
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2 x10
0.6
0.8
1
1.2
1.4
1.6
1.8
2
2.2
6.7 6.8 6.9 7 7.1 7.2 7.3 7.4 7.5 7.6 7.7
2 x10
0.6
0.7
0.8
0.9
1
1.1
1.2
6.5 6.6 6.7 6.8 6.9 7 7.1 7.2 7.3 7.4 7.5
2 x10
0.5
0.6
0.7
0.8
0.9
1
1.1
1.2
1.3
1.4
5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1 6.2 6.3
2 x10
1.75
2
2.25
2.5
2.75
3
3.25
3.5
3.75
4
4.25
5.2 5.3 5.4 5.5 5.6 5.7 5.8 5.9
2 x10
0.5
0.75
1
1.25
1.5
1.75
2
2.25
2.5
2.75
3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9
2 x10
1
2
3
4
5
6
7
2.1 2.2 2.3 2.4 2.5 2.6
Retention time (min)
RR 930 pg/mL
YR 107 pg/mL
LR 155 pg/mL
LY 292 pg/mL
LW 176 pg/mL
LF 195 pg/mL
LA 179 pg/mL 2
x10
0.5
0.6
0.7
0.8
0.9
1
1.1
1.2
1.3
1.4
3 3.1 3.2 3.3 3.4 3.5 3.6 3.7
• The highly sensitive 6495 allows confident quantitation of microcystins at pg/mL levels in liver
extract samples
Summary of Microcystins Quantitation Excellent linearity, precision and accuracy at all levels
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Compounds LLOQ
(pg/mL)
Range
(pg/mL) R2 Precision
(%RSD, n = 3) %Accuracy
Liver Extract
(pg/mL)
RR 2.5 2.5 - 50,000 0.9949 0.13 - 6.24 93.9 - 111.7 930
YR 2.5 2.5 - 50,000 0.9906 1.30 - 13.51 91.6 - 111.9 107
LR 2.5 2.5 - 50,000 0.9921 0.41 - 11.35 92.2 - 107.5 155
LA 25 25 - 50,000 0.9984 0.57 - 5.40 85.8 - 101.9 179
LY 50 50 - 50,000 0.9975 2.80 – 1034 88.8 - 119.4 292
LW 25 25 - 50,000 0.9987 0.66 - 14.3 85.6 - 105.1 176
LF 25 25 - 50,000 0.9975 1.62 - 6.35 85.5 - 115.0 195
MassHunter Acquisition B.07
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Acquisition
Quant
Support for 6495 QQQ LC/MS
Faster and accurate autotune
Improved workflows
Optimizer to better support tMRM
Dynamic MRM (dMRM) and triggered MRM (tMRM)
iFunnel express, Study manager, Skyline automation, application kits and solutions
Fast and Accurate Automatic Tuning Improved System Robustness
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• New autotune reduces the time by 40 min (40%) when both POS & NEG polarities are tuned
• New checktune reduces the time by 2 min (25%) when both polarities are checktuned
• Enhanced quadrupole optics tuning algorithms results in higher ion transmission
Shortens 20 min
for each polarity
Dynamic MRM (dMRM)
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Dynamic MRM, Fast MRM Speed and Polarity Switch
• Group MRMs in RT windows
instead of time segments
• 2x shorter cycle time supports
narrow UHPLC peaks
• Supported by very fast MRM
speed (1 ms dwell) and
polarity switch (20 ms)
Triggered MRM (tMRM)
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• tMRM database accelerates
acquisition method setup
• tMRM library – Match Score
for confirmation
• tMRM is Fast and Sensitive
• UHPLC compatible
• Simultaneous Quantitation,
Screening and Confirmation
Triggered MRM, tMRM Database and Library
Match
Score
95.1 Trigger
Threshold
tMRM Product
Ion Spectrum
Apex
Enhanced Optimizer Workflow in MassHunter B.07 (to Better Support tMRM)
• Find 10 MRM transitions in Optimizer
• Injection volume flexibility
• Improved method editing
MassHunter
Optimizer B.07
iFunnel Express iFunnel Express – Source and iFunnel Optimization
Method Generation Acquisition Optimization
• Automate the source/iFunnel
optimization process
Create
Methods
& Worklist
MH Source
and Funnel
Optimizer
Acquire Data
for Each
Parameter
MassHunter
Quant
Data
Files Integrate for
Peak Data
Excel File for
Optimized
Parameter
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Study Manager and Skyline Automation
Skyline Automation
Study Manager – Fully Automated Quantitation Workflow
1. Specify input File 2. Specify Quant Method 3. Specify Report 4. Submit and go!
MH Acquisition MH Quant
• Automated workflows for peptide quantitation
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LC/MS Applicatio
n Kits
Method
Standards
On Site Trainin
g
LC Column
tMRM Databas
e & Library
tMRM Application Kits For LC/MS Targeted Screening & Confirmation with QQQ
Pesticides
Test Mix: 254 compounds
DB: 700+ compounds
Library: 200+ compounds
Veterinary Drugs
Test Mix: 146 compounds
DB: 500+ compounds
Library: 100+ compounds
Forensic Toxicology
Test Mix: 139 compounds
DB: 2500+ compounds
Library: 100+ compounds
• Agilent unique data dependent acquisition for fast and sensitive compound
screening, quantitation and confirmation.
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Summary: Value of the New 6495 QQQ LC/MS
Increased sensitivity
Lowest Limits of Quantitation
Reproducible, High Quality Data
- More Confident Results
Ease of Operation & Fast Set-up
- Get Answers Quickly!
Screen, Confirm and Quantify
with UHPLC Speed
Streamlined Analytical Workflow
- More Productive Lab
Simpler Methods & Faster
Results
Less Maintenance & Downtime
Excellent precision & accuracy
at the Lowest Levels Wide linear dynamic range
Fast MRM speed, dynamic and
triggered MRM
Ease to use, workflow driven
software, kits and solutions
Proven robustness and
stability in complex matrices
Sample dilution
Direct injection, LC…
Confident quantitation and confirmation for the most demanding applications
6495 Brochure, Video, and Application Notes
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6495 Brochure: 5991-4541EN
6495 Video
6495 App Note: 5991-4687EN
6495 App Note: 5991-4686EN
6495 App Note: 5991-4685EN
ASMS 2014, Baltimore Celebrating Achievement
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