Agilent New 6470 Triple Quadrupole LC/MS System

Transcription

1 Agilent New 6470 Triple Quadrupole LC/MS System ROCK SOLID Performance For Confident Quantitation Na Pi Parra, Ph.D Product Manager for Triple Quadrupole LC/MS 1

2 Overview of Topics The New 6470 QQQ LC/MS System Technology innovations Quantitation performance Key Applications Food safety Environmental Pharmaceutical Summary 2

3 Introducing the 6470 Triple Quadrupole LC/MS Rock Solid Performance Improved Sensitivity Lower IDL < 4.0 fg Smaller Footprint Upgradeable to 6495 Higher Throughput Higher Confidence Higher Productivity

4 6470 QQQ Technologies Enhanced Performance in a Smaller Space Agilent Jet Stream Technology Thermal gradient focusing Efficient desolvation Creates an ion rich zone Up to 10x gains in sensitivity 3 2 An Ion Detector with High Energy Conversion Dynode and Low Noise Improved ion detection A Curved and Tapered Hexapole Collision Cell Effective ion collection 1 Enhanced Q1 Ion Optics Improved ion transmission 4

5 Enhanced Q1 Ion Optics with Optimized Prefilter Improved transmission of precursor ions and additional system robustness 1. Reduced Octopole length 21. New prefilter 2 1 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 More reliable and robust performance 5

6 Curved and Tapered Hexapole Collision Cell Effective collection and transmission of product ions and smaller footprint 6460 QQQ LC/MS - Linear Collision Cell 30% smaller 6470 QQQ LC/MS - Curved Collision Cell Curved and Tapered Hexapole Assembly for efficient collection and transmission of product ions Designed for consistent collision energies across all QQQ platforms A compact, smaller benchtop footprint 6

7 New Detector with High Energy Conversion Dynode More efficient detection and quantitation of ions with low noise characteristics 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 mass range Low noise level at 20 kv % increase in response to 20kV vs. 10kV - Improved signal to noise 7

8 Performance 6470 QQQ Rock Solid, Improved Performance Robust and Reliable, More Sensitive, More Compact, and Upgradeable Robustness Proven robustness and reliability in complex matrix - Extensively tested in key applications Improved sensitivity and lower LLOQs In S/N specifications and key applications Improved precision and excellent accuracy In IDL specifications and key applications Improved scan and MRM speed, and polarity switching Proven 6 orders of linear dynamic range Footprint Smaller (30%) footprint and upgradeable to the

9 Unprecedented Robustness in Biological Matrix Quantitation of Drugs in Plasma using RapidFire QQQ Love et al. WP #496 Injection #1 Injection #10,000 Time (Sec) Injection #1 Injection #10,000 Time (Sec) Stable peak area response over 10,000 injections of plasma samples and 5 days of continuous operation Response %RSD =

10 Improved Sensitivity and Precision Lower IDLs Reserpine (+) and Chloramphenicol (-), 6470 vs Models Reserpine (+) Chloramphenicol (-) Measured at %RSD IDL Measured at %RSD IDL fg fg 20 fg fg fg fg 10 fg fg 10 fg reserpine (+) on the fg chloramphenicol (-) on the 6470 IDL = 2.9 fg %RSD = 10.2 n = 10 injections IDL = 2.5 fg %RSD = 8.7 n = 10 injections 6470 IDLs are measured at a 2x lower level 6470 achieves 2-3x lower IDLs vs. the

11 Improved Sensitivity and Precision Lower LLOQ Acetaminophen LLOQ in Water, 6470 vs LLOQ = 2 ng/l on the 6460 LLOQ = 0.5 ng/l on the 6470 n = 8 injections 6470 Peak Area = 5863 S/N = 34.6 Area %RSD = 4.3 n = 8 injections 6470 Peak Area = 1656 S/N = 10.6 Area %RSD = 8.3 %Accuracy = Peak Area = 1490 S/N = 11.1 Area %RSD = 9.5 Blank 6460 Peak Area = 423 S/N = 4.0 Area %RSD = 19.0 % Accuracy = 77.0 Improved peak area (4x) and area %RSD (2x) were observed on the 6470 compared to the 6460 Lower LLOQ (4x) was observed on the 6470 compared to the 6460 Excellent precision (%RSD) and accuracy were achieved at the LLOQ level of

12 Excellent Precision at the Lowest Levels Lower IDL Acetaminophen IDL measured at the LLOQ level, 6470 vs Models Amount measured - at LLOQ Replicates %RSD IDL ng/l n = ng/l ng/l n = 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 Inj # Peak Area S/N %RSD 8.3 Ave ng/l 0.5 ng/l Acetaminophen 6470 IDL = 0.1 ng/l 6460 IDL = 0.5 ng/l Lower LLOQ (4x) and IDL (5x) were achieved on the 6470 compared to the 6460 Blank 12

13 Attogram Sensitivity (+) 500 attograms of Clenbuterol on-column Area %RSD = 12.4 n = 3 injections %Accuracy = S/N = 5.9 LLOQ of Clenbuterol on the attogram on-column Blank Excellent precision (%RSD) and accuracy are observed at the LLOQ level of

14 Attogram Sensitivity (-) 800 attograms of Chloramphenicol on-column Area %RSD = 18.1 n = 3 injections %Accuracy = 98.2 LLOQ of Chloramphenicol on the attogram on-column Blank S/N = 18.0 Excellent precision (%RSD) and accuracy are observed at the LLOQ level of

15 5.7 Orders of Linear Dynamic Range Clenbuterol (+), 0.1 pg/ml 50 ng/ml Excellent linearity, R 2 = Superb assay accuracy and precision are achieved at all levels including the LLOQ level of 0.1 pg/ml Levels %RSD (n = 3) % Accuracy 0.1 pg/ml pg/ml pg/ml pg/ml pg/ml pg/ml ng/ml ng/ml

16 Applications for 6470 QQQ LC/MS Food Safety Analysis of multi-residue pesticides in food matrices with higher degrees of sample dilution Environmental Determination of PPCPs in surface water using direct injection Pharmaceutical High-Capacity Multisampler UHPLC High-throughput analysis of drugs & metabolites in plasma with reliable 24/7 operation 16

17 Multi-Residue Pesticide Analysis in Food 1:20 dilution of > 250 pesticides spiked into black tea at 10 ng/ml (ppb) Injection volume = 2 µl Sartain et al. WP #059 Multi-residue pesticide analysis in food products most demanding food safety applications Improved sensitivity and precision of the 6470 allows accurate quantitation of pesticides <Maximum Residue Limits (MRLs) imposed by EU, with higher degrees of sample dilution Sample dilution reduces matrix effects, improves method robustness, allows more efficient ionization and enables the use of solvent calibration with better accuracy 17

18 Sensitivity Improvements for High Relevance Pesticides 1:20 Dilution of 10 ng/ml of Pesticides in Black Tea, Peak Area 6470 vs Area Gain Oxamyl 2.8x Area Gain Acetamiprid 3.4x Methomyl 3.9x Thiacloprid 2.7x Thiamethoxam 2.5x Ethion 3.2x Improved sensitivity (average peak area gain of 3.0x) are observed on the new 6470 vs

19 Improved Precision and Lower IDLs IDLs of 30 Highly Relevant Pesticides, 6470 vs Pesticide IDL (ppt) Fold- Pesticide IDL (ppt) Fold Improvement Improvement Acephate x Fipronil x Acetamiprid x Flufenoxuron x Azinphos-methyl x Imazalil x Buprofezin x Imidacloprid x Carbendazim x Isocarbophos x Chloroxuron x Metamitron x Cycluron x Methidathion x Desmedipham x Methomyl x Diethofencarb x Monocrotophos x Dimethoate x Oxamyl x Dimethomorph x Pirimicarb x Dimoxystrobin x Pyridaben x Diniconazole x Tebuconazole x Dioxacarb x Teflubenzuron x Diuron x Triazophos x Average Improvement 3.6X Improved precision (peak area %RSD) are observed, particularly at the lowest levels The enhanced peak area response and improved precision (%RSD) leads to lower LLOQs and IDLs compared to the 6460 Lower IDLs (average improvement of 3.6x) was observed across all > 250 pesticides 19

20 Femtogram Sensitivity for Pesticides in Black Tea LLOQs and IDLs / MDLs at Low-pg/mL level on the 6470 Pesticides Oxamyl (+) Pirimicarb (+) Bentazone (-) Flubendiamide (-) Neat Solvent %Accuracy %RSD (n = 5) LLOQ (pg/ml) IDL (pg/ml) Black Tea %Accuracy %RSD (n = 5) LLOQ (pg/ml) MDL (pg/ml) Oxamyl 10 pg/ml S/N = 10.4 Bentazone 10 pg/ml S/N = 17.6 LLOQs in Black Tea Pirimicarb 10 pg/ml S/N = 12.7 Flubendiamide 20 pg/ml S/N = 195 LLOQs 100 pg/ml are required to achieve the MRL with 1:20 dilution of black tea Excellent peak area precision (%RSD) and accuracy are observed at the LLOQs in black tea 20

21 Improved Sensitivity and Precision of the 6470 Allows for Higher Degree of Sample Dilution Histograms of LLOQs for Evaluated Pesticides in Solvent and Food Matrices Tomato Orange Black Tea ppt ppt ppt 500 ppt - 1 ppb 1-10 ppb >10 ppb Majority of the pesticides achieved an LLOQ of 5% of the MRL level in food matrices Pesticides can be confidently quantified in food matrices with up to 1:20 dilution Sample dilution leads to reduced matrix effects and improved method robustness Sample dilution allows for more efficient ionization and significantly better recoveries 21

22 Number of Pesticides Improved Recoveries at Higher Degree of Dilution undiluted 1 to 2 1 to 5 1 to 10 1 to 20 Passes SANCO Requirements 0 <30% 30-50% 50-70% % >120% Recoveries 116 pesticides showed strong ion suppression in undiluted black tea Most of pesticides achieved acceptable recoveries of % and less signal suppression with 1:20 dilution, which are in full compliance with SANCO requirements Neat solvent calibration curve can be used for pesticide quantitation with better accuracies 22

23 Applications for 6470 QQQ LC/MS Food Safety Analysis of multi-residue pesticides in food matrices with higher degrees of sample dilution Environmental Determination of PPCPs in surface water using direct injection Pharmaceutical High-Capacity Multisampler UHPLC High-throughput analysis of drugs & metabolites in plasma with reliable 24/7 operation 23

24 Determination of PPCPs in Environmental Water 24X IN POS Mode Erythromycin (+) Acetaminophen (+) Fluridone (+) Atenolol (+) Gabapentin (+) Atrazine (+) Lamotrigine (+) Bupropion (+) Metoprolol (+) Caffeine (+) Propranolol (+) Carbamazepine (+) Sucralose (+) Clarithromycin (+) Sulfamethoxazole (+) Cotinine (+) Trimethoprim (+) DEET (+) Venlafaxine (+) Dextrorphan (+) 4x in NEG Mode Diazinon (+) 2,4-D (-) Diltiazem (+) Gemfibrozil (-) Diphenhydramine (+) Triclopyr (-) Diuron (+) Triclosan (-) 28 PPCPs spiked in water at 10 ng/l (ppt) Injection volume = 40 µl Li et al. TP #305 PPCPs in surface waters may have an adverse impact on wildlife and human even at very low concentrations (ng/l) The improved sensitivity and precision of the 6470 allows accurate quantitation of PPCPs at sub-ng/l level using direct injection without the need of time-consuming SPE enrichment Direct injection improves analysis efficiency, assay accuracy, and lab productivity 24

25 Improved Sensitivity for PPCPs in Positive Mode 10 ng/l of PPCPs in Water, Peak Area 6470 vs Acetaminophen 2.6x 6470 Bupropion 3.5x Clarithromycin 4.5x Dextrorphan 1.8x 6460 Atenolol 2.3x Caffeine 2.4x Cotinine 2.9x Diazinon 2.6x Atrazine 3.3x Carbamazepine 2.2x DEET 2.6x Diltiazem 2.1x Improved sensitivity (average peak area gain of 2.9x) are observed across 24 PPCPs in POS mode 25

26 Improved Sensitivity for PPCPs in Negative Mode 10 ng/l of PPCPs in Water, Peak Area 6470 vs , 4-D 4.1x Gemfibrozil 2.3x Triclopyr 3.8x Triclosan 2.7x Improved sensitivity (average peak area gain of 3.2x) are observed across 4 PPCPs in NEG mode Overlaid MRM chromatograms: 6470 vs ppt on ppt on x in POS compounds 3.2x for NEG compounds Average improvement = 3.0x 26

27 Improved Precision Comparison of Area %RSD 5 ng/l of PPCPs in Water, 6470 vs Compounds Improved precision (peak area %RSD) are observed, particularly at the lowest levels Average improvement of 2.3x was observed across 28 PPCPs analyzed %RSD (n=8) Improvement Compounds %RSD (n=8) Improvement Acetaminophen 0.6x Erythromycin 1.3x Atenolol 1.6x Fluridone 6.0x Atrazine 2.0x Gabapentin 1.7x Bupropion 2.7x Lamotrigine 3.3x Caffeine 3.6x Metoprolol 1.3x Carbamazepine 4.9x Propranolol 1.8x Clarithromycin 1.3x Sucralose 1.2x Cotinine 3.0x Sulfamethoxazole 1.6x DEET 2.2x Trimethoprim 2.1x Dextrorphan 1.8x Venlafaxine 2.0x Diazinon 2.0x 2,4-D 2.1x Diltiazem 1.4x Gemfibrozil 3.6x Diphenhydramine 4.0x Triclopyr 2.4x Diuron 1.0x Triclosan 1.2x Average Improvement 2.3x The enhanced peak area response and improved precision (%RSD) leads to lower IDLs and LLOQs compared to the

28 Lower IDLs at Sub-ng/L Level IDLs of PPCPs in water, 6470 vs IDLs of 24 PPCPs in Positive Mode 6470 IDL 6460 IDL IDLs of 4 PPCPs in Negative Mode 6470 IDL 6460 IDL PPCPs 6460 IDL 6470 IDL Improve (ng/l) (ng/l) ment PPCPs 6460 IDL (ng/l) 6470 IDL (ng/l) Improve ment Acetaminophen x Erythromycin x Atenolol x Fluridone x Atrazine x Gabapentin x Bupropion x Lamotrigine x Caffeine x Metoprolol x Carbamazepine x Propranolol x Clarithromycin x Sucralose x Cotinine x Sulfamethoxazole x DEET x Trimethoprim x Dextrorphan x Venlafaxine x Diazinon x 2,4-D x Diltiazem x Gemfibrozil x Diphenhydramine x Triclopyr x Diuron x Triclosan x Average Improvement 3.6x Lower IDLs (average improvement of 3.6x) were observed across all 28 PPCPs (POS and NEG) 28

29 Lower LLOQs Femtogram Sensitivity for PPCPs LLOQs of PPCPs in water, 6470 vs PPCPs 6470 LLOQ (ng/l) 6460 LLOQ (ng/l) PPCPs 6470 LLOQ (ng/l) 6460 LLOQ (ng/l) Acetaminophen Erythromycin Atenolol Fluridone < Atrazine Gabapentin Bupropion Lamotrigine Caffeine Metoprolol Carbamazepine Propranolol Clarithromycin Sucralose Cotinine Sulfamethoxazole DEET Trimethoprim Dextrorphan Venlafaxine Diazinon ,4-D Diltiazem Gemfibrozil Diphenhydramine Triclopyr Diuron Triclosan Improvement 2 5 X Acetaminophen 0.5 ng/l %RSD = 8.3 (n=8) %Accuracy = S/N = 22 Bupropion 0.2 ng/l %RSD = 7.3 (n=8) %Accuracy = 99.0 S/N = 35.1 Fluridone 0.1 ng/l %RSD = 3.8 (n=8) %Accuracy = S/N = 66 Triclosan 20 ng/l %RSD = 8.9 (n=8) %Accuracy = S/N = 33.4 Lower LLOQs (average improvement of 2.7x) were achieved across all 28 PPCPs (POS and NEG) LLOQs for most of the PPCPs are much lower than the ng/l (ppt) level regulation limit Excellent peak area precision (%RSD) and accuracy are observed at the LLOQs 29

30 Applications for 6470 QQQ LC/MS Food Safety Analysis of multi-residue pesticides in food matrices with higher degrees of sample dilution Environmental Determination of PPCPs in surface water using direct injection Pharmaceutical High-Capacity Multisampler UHPLC High-throughput analysis of drugs & metabolites in plasma with reliable 24/7 operation 30

31 Analysis of Drugs and Metabolites in Human Plasma 6470 QQQ with High-Capacity Multisampler UHPLC Benzodiazepines and metabolites spiked in human plasma at 5 pg/ml Midazolam Yang et al. Midazolam -OH Midazolam -Hydroxymidazolam TP #297 Alprazolam - Hydroxyalprazolam Alprazolam -OH Alprazolam PK analysis of drugs and metabolites is challenged with the need to perform large-scale-analyses with high sensitivity (pm) and sufficient dynamic range (pm nm) in biological matrices The highly sensitive 6470 enables fast and accurate quantitation of drugs and metabolites at low-pm level over a wide linear dynamic range in human plasma 6470 s robustness together with the high-capacity multisampler allows for high-throughput drug analysis with 24/7 unattended operation and extremely reliable results 31

32 High Sensitivity for Benzodiazepines in Plasma LLOQs and MDLs at Low-pg/mL level on the 6470 Compound LLOQ (pg/ml) %Accuracy %RSD (n = 7) MDL (pg/ml) -Hydroxymidazolam Midazolam Hydroxyalprazolam Alprazolam Hydroxymidazolam LLOQ = 5 pg/ml S/N = Hydroxyalprazolam LLOQ = 5 pg/ml S/N = 5.4 Midazolam LLOQ = 5 pg/ml S/N = 35.2 Alprazolam LLOQ = 5 pg/ml S/N = 9.2 Excellent peak area precision (%RSD) and accuracy are observed at the LLOQ level 32

33 Relative Responses Relative Responses Confident Quantitation over 5 Orders of Dynamic Range x E-05 1E-05 5E-06 Midazolam, 5 pg/ml 500 ng/ml 5 Orders of Linear Dynamic Range R 2 = Concentration (ng/ml) Levels %RSD (n = 7) %Accuracy 5 pg/ml pg/ml pg/ml pg/ml pg/ml ng/ml ng/ml ng/ml ng/ml ng/ml ng/ml x E-05 1E-05 5E-06 Alprazolam, 5 pg/ml 500 ng/ml 5 Orders of Linear Dynamic Range R 2 = Concentration (ng/ml) Levels %RSD (n = 7) %Accuracy 5 pg/ml pg/ml pg/ml pg/ml pg/ml ng/ml ng/ml ng/ml ng/ml ng/ml ng/ml Excellent linearity (R 2 > 0.995), accuracy and precision are achieved at all levels including LLOQ Drugs and metabolites at different levels can be quantified simultaneously and in one experiment 33

34 Peak Area Ratio Peak Area Ratio Peak Area Ratio Peak Area Ratio Proven System Robustness in Human Plasma -Hydroxymidazolam Midazolam Area Ratio %RSD=1.3 Peak area %RSD= Area Ratio %RSD=3.3 Peak area %RSD= Injection number Injection number -Hydroxyalprazolam Alprazolam Area Ratio %RSD=1.4 Peak area %RSD= Area Ratio %RSD=1.3 Peak area %RSD= Injection number Injection number Stable peak area response was observed over 1,300 injections of plasma extract and 1 week of continuous operation Average area ratio %RSD = 1.8 was observed with use of internal standard (ISTD) Average peak area %RSD = 6.6 was observed for benzodiazepines and metabolites 34

35 Summary: 6470 QQQ LC/MS Rock Solid Performance for Confident Quantitation and Highest Lab Productivity Additional Robustness Less Maintenance Cleaning Reliable Consistent Results over Longer Time (24/7) Upgradeable to the 6495 Ion Optics Improved Sensitivity Streamlined Analytical Workflow 20KV Detector Reproducible High Quality Data Curved Collision Cell Higher Throughput at UHPLC Speed Smaller Footprint Improved Precision More Instruments In Your Laboratory Faster Speed Protection of Your Investment 35

36 ASMS 2015, St Louis 6470 Marketing Launch Breakfast Seminar Agilent Booth Technical VIP Hospitality Suite Technical Posters Brochure App Note 36

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