Quantitative Analysis of pesticides including Phenoxy-acids in surface and drinking water matrices using UPLC-ON-LINE-SPE-MS/MS

Transcription

1 Quantitative Analysis of pesticides including Phenoxy-acids in surface and drinking water matrices using UPLC-ON-LINE-SPE-MS/MS Alexandre Paccou Demo Centers and Application Support Manager, Southern Europe Food & Environment Market Development Manager, Sothern Europe Milan 12/03/ Waters Corporation 1

2 Waters UPLC with On-line SPE Technology 2013 Waters Corporation 2

3 Off Line Sample Preparation Extraction Protocol Barrels or Syringe Prepare Sample Condition/Equilibrate 3 ml methanol/3 ml water Load > 500 ml sample Wash Elute OFF-line SPE Extraction Manifold For GC analysis, dry extract over Na 2 SO 4, then adjust to 1mL For LC analysis, exchange to acetonitrile, then adjust to 1 ml 2013 Waters Corporation 3

4 Off Line Sample Preparation Extraction Protocol Volume Availability 1 ul 50 ul 100 ul 500 ul 1 ml 5 ml 10 ml 50 ml 100 ml 500 ml 1 L Condition/Equilibrate Load wash 10 x 100 x 1000 x Elute Evaporate and Reconstitute Final extract volume 1mL inject µl 1000x 100x 10 x Method LOQ Target LOQ 1 ng/l 10 ng/l 100 ng/l 1 ug/l 10 ug/l 100 ug/l 1 mg/l 10 mg/l 100 mg/l 1 g/l 10 g/l Instrument LOQ 2013 Waters Corporation 4

5 Drinking Water Analysis How It s Done Currently Collection Sample Preparation Analysis Results - Filtration - Extraction - Clean up - Concentration - Derivatization LC/MS GC/MS LC/MS/MS GC/MS/MS Qualitative Quantitative RSD s > 15 % + 1L to 20 L - Sample container - Shipping container - Manual Work - Hours to days - Consumable Disposable - Sample storage - Lab space - Additional Equipment: - Glass ware - Manifold - Evaporator Automated System - Shipping cost 2013 Waters Corporation 5

6 ON Line SPE/LC/MS/MS ON-line analysis is a technique that reduces the amount of manual labor to produce an analyzable sample for LC/MS/MS. The sample is injected on an extraction column using an automated injector. Barrels or Syringe OFF-line SPE Extraction Manifold 2013 Waters Corporation 6

7 Volume Availability 1 ul 50 ul 100 ul 500 ul 1 ml 5 ml 10 ml 50 ml 100 ml 500 ml 1 L Condition/Equilibrate Load wash Elute Final extract volume is 0 (ml, L, nl?) Analytes are trapped on Extraction Column Direct elution to HPLC Column Off Line SPE Extraction LC/MS Analysis 500 ml (starting volume) 1 ml final (elution volume) µl (injection volume) 500:1 enrichment 30 µl 1/33 th of original volume 15.0 ml 50 µl 1/20 th of original volume 25.0 ml 2013 Waters Corporation 7

8 Drinking Water Analysis Going Forward: ON-line SPE/LC/MS/MS System Collection Sample Preparation Analysis Results 20 ml - Sample container - Shipping container ON-line SPE/LC/MS/MS Integrated System - Fully Automated no manual work - Results in minutes instead of hours/days - Reusable chemistries longer cycle time - Reduction of sample storage space - Minimum lab space required - No additional equipment needed - Reduction of analytical costs Qualitative Quantitative RSD s < 15 % - Shipping cost 2013 Waters Corporation 8

9 Traditional Off-Line Analytical Approach Collection Off-line Sample Preparation Analysis Results Most time consuming steps 2013 Waters Corporation 9

10 Challenge Complex and Labor Intensive Work Flow Large volumes of dilute samples Multiple Vessel Transfers Time Consuming Steps Separations are slow Rework can be high MS is required, skilled, expensive operators Competition between CRO s is high and Costs must be reduced 2013 Waters Corporation 10

11 Waters UPLC with Online SPE Technology 2777c or 2777 Column Heater/Cooler QSM BSM Xevo TQD Xevo TQ-S 2013 Waters Corporation 11

12 Receiving and Preparing Samples Shipping Box Pre-slit Septum Cap 20 ml vial Autosampler rack Solid cap 2013 Waters Corporation 12

13 New Analytical Approach For Analysis of Compounds of Concern in Water Collection On-line Sample Preparation Analysis Results 20 ml Sample concentration Elution Analysis 2013 Waters Corporation 13

14 System Diagram A: injection valve for online SPE UPLC B: switching valves for extraction columns C: injection valve for stand-alone UPLC Parallel extraction columns increase throughput Simple switch between online SPE system and stand alone UPLC system 2013 Waters Corporation 14

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22 Example events 2013 Waters Corporation 22

23 Online SPE Method Fundamentals Loading Eluting Basic 100 % aqueous + 2 % NH 4 OH ph 10 High K - basic functionality neutral - highest trapping efficiency Organic gradient 1, 3 or 5 min % acidic additive (ph 3-5) FA, AA, NH 4 Formate or NH 4 Acetate Low K - basic functionality ionized Neutral 100 % aqueous + no additive Organic gradient 1, 3 or 5 min with or without additive (ionisation may improve with additive) Acidic 100 % aqueous + 2 % FA ph 3 High K - acidic functionality neutral - highest trapping efficiency Organic gradient 1, 3 or 5 min % basic additive (ph 10) NH 4 OH or NH 4 HCO 3 Low K - acidic functionality ionized 2013 Waters Corporation 23

24 Loading and Eluting Conditions Load High K 100% Aqueous Backflush Elution SPE UPLC SPE Sorbents: Oasis HLB 20 m ph 1-14 XBridge C18 10 mph 1-12 XBridge C8 10 mph 1-12 Waste Elute Low K ESI + MS/MS 2013 Waters Corporation. 24

25 Recommended Columns UPLC packed online SPE direct connect columns: BEH columns are recommended for elution ph tolerance Waters Corporation 25

26 Analysis of Pesticides in ESI Waters Corporation 26

27 Analysis of Pesticides in ESI Waters Corporation 27

28 Analysis of Phenoxy-acids pesticides in ESI Waters Corporation 28

29 Analysis of Phenoxy-acids pesticides in ESI Waters Corporation 29

30 Linearity Example 10 to 150 ppt 2013 Waters Corporation 30

31 Linearity Example 10 to 150 ppt 2013 Waters Corporation 31

32 Sensitivity ESI+ 10ppt 2013 Waters Corporation 32

33 Sensitivity ESI- 10ppt 2013 Waters Corporation 33

34 LOD/LOQ 2013 Waters Corporation 34

35 Repeatability 2013 Waters Corporation 35

36 % % % Lifetime Metolachlor 500 th injection MRM of 15 Channels ES > e MRM of 15 Channels ES > e5 250 th injection MRM of 15 Channels ES > e5 1st injection Time 2013 Waters Corporation 36

37 UPLC with On-line SPE Technology for Complex Matrices Collection Off-line Sample Preparation On-line Sample Concentration Analysis Results 20 ml Sample concentration Elution Analysis 2013 Waters Corporation 37

38 UPLC with On-line SPE Technology Sensitivity o Range of solutions addressing ppm to low ppt analyte levels Robustness o Column lifetime in excess of 500 injections for Drinking Water Flexibility o o Operate as On-line SPE and Standalone Multiple Detectors Reproducibility o RSDs always less than 15% Transferability/Ease of Use o Advance software suite 2013 Waters Corporation 38

39 Conclusion A UPLC-on-line-SPE-MS/MS method for the analysis of pesticides in Surface and Drinking Water samples at ppt level was developed On-line SPE and UPLC parameters were optimized in order to be able to analyze the widest range possible of pesticides Only a single 5mL Water Sample injection was required for this analysis A throughput of 3 to 4 samples per hours including preparation was typically achieved Apart from an eventual filtration, no sample pre-treatment is needed, An excellent linearity was observed across the studied range of 10 to 150ppt with coefficient of determination higher than for all studied compounds Standard deviation from the calibration curve always stayed below 10% even at 10ppt An excellent sensitivity was observed for all compounds Calculated detection limits ranged from below 0.05 to ppt Repeatability studies revealed Rsds typically lower than 10% for all compounds even at low level Excellent column life-time > 500 injections 2013 Waters Corporation 39

40 Thank You! 2013 Waters Corporation 40