FOOD TESTING ACQUITY QDa Simplify matrix complexity and extend detection capabilities
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1 FOOD TESTING ACQUITY QDa Simplify matrix complexity and extend detection capabilities
2 LC-MS 3 ACQUITY UPLC H-Class ACQUITY QDa 13 ACQUITY UPLC H-Class ACQUITY QDa 19 ACQUITY UPLC H-Class /ACQUITY QDa 23 ACQUITY QDa 28 Myco6in1 + ACQUITY QDa ACQUITY UPLC H-Class ACQUITY QDa 38 ACQUITY UPLC I-Class /ACQUITY QDa 42 Alliance HPLC
3 ACQUITY UPLC H-Class ACQUITY QDa Mark E. Benvenuti, Dimple Shah, and Jennifer A. Burgess Waters Corporation, Milford, MA, USA MS ACQUITY QDa UPLC HPLC UV ACQUITY QDa LC EC 1925/2006 CFR 21 HPLC 1 LC-MS ACQUITY QDa 12 WSV ACQUITY UPLC H-Class ACQUITY UPLC QDa ACQUITY UPLC H-Class ACQUITY QDa WSV QDa 3
4 1 WSV SIR m/z Analyte RT (Min) SIR m/z Cone voltage (V) Ascorbic Acid (C) Thiamine (B1) Nicotinic Acid (B3) Pyridoxal (B6) Nicotinamide (B3) Pyridoxine (B6) Ca_Pantothenate (B5) Cyanocobalamin (B12) Folic Acid (B9) B2-5-Phosphate Biotin (B7) Riboflavin (B2) SIR Standard Individual B complex vitamins (mg/l ) Vitamin C (mg/l) WSV 1 mg/ml B2 B7 B9 1N NaOH 200 µl C ph C 1.25 ml ml 25 ml C 50 ppm 1 ppm g 100 ml 0.2 µm PVDF g 100 ml µm PVDF µm PVDF 2. B C 4 ACQUITY UPLC H-ClassACQUITY QDa
5 UPLC LC ACQUITY UPLC H-Class 17.5 ACQUITY UPLC HSS T3 1.8 µm mm A 10 mm 0.1% B 10 mm 0.1% 5 µl ml/ %A %B 1. Initial ACQUITY UPLC PDA 210~400 nm 270 nm 10 / 2 ACQUITY QDa ESI kv MS 50~800 m/z SIR 5 Hz 15 V 1 SIR SIR m/z 2 3 ACQUITY UPLC H-ClassACQUITY QDa 5
6 MS m/z 2 2 D F m/z=442m/z =678 2 UV 2A 2H UV 3 MS Ascorbic Acid C 2- Thiamine B1 3- Nicotinic acid B3 4- Pyridoxal B6 5- Nicotinamide B3 6- Pyridoxine B6 7- Ca_Pantothenate B5 8- Cyanocobalamin B12 9- Folic acid B9 10- Riboflavin 5 phosphate 11- Biotin B7 12- Riboflavin B SIR 6 ACQUITY UPLC H-ClassACQUITY QDa
7 Compound name: Biotin (B7) Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None A Compound name: Thiamine (B1) Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None B Response Response ppb ppb Compound name: Riboflavin (B2) Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None C Compound name: Folic Acid (B9) Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None D Response Response ppb ppb Compound name: Riboflavin 5 phosphate Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None E Compound name: Cyanocobalamin ( B12) Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None F Response Response ppb Conc Compound name: Nicotinic Acid (B3) Coefficient of Determination: r 2 = Calibration curve: * x * x Response type: External Std, Area Curve type: 2nd Order, Origin: Exclude, Weighting: 1/x, Axis trans: None G Compound name: Ca_Pantothenate (B5) Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None H Response Response ppb ppb ACQUITY UPLC H-ClassACQUITY QDa 7
8 MS UV 3 5 ppb 5 µg/l SIR UV 3A 270 nm 3A UV MS 5.0e-4 AU 0.0 A: UV at 270 nm % RT 2.46, S/N = 63 B: Pyridoxine SIR m/z % RT 1.75, S/N = 32 C: Pyridoxal SIR m/z % RT 1.24, S/N = 19 D: Nicotinic Acid SIR m/z % RT 2.43, S/N = 33 E: Nicotinamide SIR m/z Time µg/ml 270 nm UV SIR SIR 4 BCD E 8 ACQUITY UPLC H-ClassACQUITY QDa
9 4 2 B5 UV UV B1 UV 1 5 B1 C C UV B1 5A ACQUITY QDa SIR B1 AU A B C % % MS Vitamin water 1 PDA Vitamin water 1 MS Vitamin water 2 7: SIR of 1 Channel ES+ 242 (Ca_Pantothenate (B5)) 14: Diode Array 7: SIR of 1 Channel ES+ 242 (Ca_Pantothenate (B5)) AU D 2.0 PDA Vitamin water 2 14: Diode Array Time 4. 2 B5 5.9 MS A CUV B D 100 Thiamine, 1.01 RT 1.01 A: SIR of thiamine (B1) % Ascorbic acid, 0.91 RT 0.91 B: SIR of ascorbic acid(c) 0% e-1 Ascorbic acid, 0.90 RT 0.90 C: UV at 270 nm AU 1.0e Time A B1 SIR B C SIR C C 270 nm UV B1 UV ACQUITY UPLC H-ClassACQUITY QDa 9
10 2 4 UPLC 6 UV 6B B12 ACQUITY QDa LC A: SIR of cyanocobalamin : SIR of 1 Channel ES+ 678 (Cyanocobalamin (B12) % B: UV at 270 nm (1) PDA Ch1 270 nm at 1.2 nm 1.0e-2 8.0e-3 AU 6.0e-3 4.0e-3 2.0e Time B12 A UV B 1 6 µg B12 16 mg B3 LC-MS 7 7A 7B B2 100 SIR 7C B12 SIR 270 nm UV 7D 12.5 ppm 41 ppb 68 % 96% 2 10 ACQUITY UPLC H-ClassACQUITY QDa
11 A 100 % Diluted x SIR at m/z 377 Riboflavin (B2) A B12 C B UV D B 100 Undiluted 7.71 SIR at m/z 377 Riboflavin (B2) % C 100 % Undiluted SIR at m/z 678 Cyanocobalamin (B12) D UV at 270nm 2.0e-1 Undiluted AU 1.0e Time B %RSD=0.6% mg/l %RSD=10% Analyte %RSD for retention time Thiamine (B1) 0.6 Nicotinic Acid (B3) 0.19 Pyridoxal (B6) 0.23 Nicotinamide (B3) 0.22 Pyridoxine (B6) 0.26 Ca_Pantothenate (B5) 0.04 Cyanocobalamin ( B12) 0.03 Folic Acid (B9) 0.03 Riboflavin 5 phosphate 0.03 Biotin (B7) 0.02 Riboflavin (B2) mg/l mg/l 10 Analyte %RSD for area Thiamine (B1) 6.78 Nicotinic Acid (B3) 2.35 Pyridoxal (B6) 2.62 Nicotinamide (B3) 2.24 Pyridoxine (B6) 2.65 Ca_Pantothenate (B5) 4.60 Cyanocobalamin ( B12) 7.00 Folic Acid (B9) Riboflavin 5 phosphate Biotin (B7) 2.77 Riboflavin (B2) mg/l 10 B ACQUITY UPLC H-ClassACQUITY QDa 11
12 UV ACQUITY QDa SIR ACQUITY QDa 1. G F M Ball (Editor). Bioavailabilty and Analysis of Vitamins in Foods. Chapman and Hall. (1998). Ch2, pp E Riches. The Rapid Simultaneous Analysis of 12 Water Soluble Vitamin Compounds. Waters Application Note No en, June, Heudi et al. Separation of water-soluble vitamins by reversed-phase high performance liquid chromatography with ultra-violet detection: Application to polyvitaminated premixes. J Chrom A. (2005) 1070: 49 56, pp AOAC Method : Vitamin B12 in Infant Formula and Adult Nutritionals. (2012). UV LC-MS Empower 3 Mass Lynx 2 LC ACQUITY QDa TEL FAX F TEL FAX Waters ACQUITY UPLC UPLC Empower MassLynx QDa The Science of What s Possible Waters Corporation 2014 Waters Corporation. Printed in Japan JA PDF 12
13 ACQUITY UPLC H-Class ACQUITY QDa Mark Benvenuti, Dimple Shah, and Jennifer A. Burgess Waters Corporation, Milford, MA, USA ACQUITY QDa 10 MS ACQUITY QDa UV K FDA 1 A 2010 EFSA EU94/35/EC 4 96/83/EC 2003/115/EC 2006/52/EC 2009/163/ EU EU 1129/2011 ACQUITY UPLC H-Class ACQUITY QDa MassLynx MS K QDa HPLC UV K UV MS 1 ACQUITY QDa MS ACQUITY QDa 1 Waters UPLC H-Class ACQUITY QDa 13
14 UPLC UPLC ACQUITY UPLC H-Class ACQUITY UPLC HSS T mm 40 2 µl 0.4 ml/min A 0.1% B 0.1% 50/50 / v/v mg/l g 100 ml /90 / v/v 10/90 / v/v g1.244 g 100 ml UPLC Time Flow rate (min) (ml/min) %A %B Curve Initial ACQUITY QDa ESI kv 5 / ACQUITY UPLC PDA nm 214 nm 254 nm 10 / m/z g 100 ml 0.22 µm PVDF ACQUITY QDa SIR m/z 2 2. SIR m/z Compounds SIR m/z Cone Voltage (V) RT (min) Acesulfame K Aspertame Cyclamate Neotame Rebaudioside A Saccharin Stevioside Sucralose SIR 5 40 V 2 14 ACQUITY UPLC H-Class ACQUITY QDa
15 Sodium Cyclamate Saccharin Sucralose Aspartame Acesulfame K Stevioside Neotame Rebaudioside A (Reb A) 1. 2 SIR A m/z 293 m/z 395 MS A m/z m/z 803.3ACQUITY QDa Acesulfame K, RT 1.04 SIR m/z : 162 Sucralose, RT 3.20 SIR m/z : 395 Saccharin, RT 1.34 SIR m/z : 182 Rebaudioside A, RT 4.11 SIR m/z : Cyclamate, RT 1.77 SIR m/z : 178 Stevioside, RT 4.14 SIR m/z : Aspartame, RT 3.20 SIR m/z : 293 Neotame, RT 4.43 SIR m/z : SIR ACQUITY UPLC H-Class ACQUITY QDa 15
16 2 3 MS MS Compound name: Reb A Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None A 4000 Response mg/l Compound name: Stevioside Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None B Response mg/l 3. A B UV UV MS UV ACQUITY QDa mg/ L 254 nm UV SIR A : Sucralose SIR m/z 395 B: Cyclamate SIR m/z 178 C : UV at254 nm 4. 5 mg/l SIR A B 254 nm UV C 16 ACQUITY UPLC H-Class ACQUITY QDa
17 R R K 98 mg/kg 15,080 mg/kg 3798 mg/kg 2 A3105 mg/kg 630 mg/kg MS 5 2 A SIR 3. 8 R2 Sweeteners Conc. range (mg/l) Correlation coefficient R 2 Acesulfame K Aspartame Cyclamate Neotame Rebaudioside A Saccharin Stevioside Sucralose mg/l 489 mg/l 145 mg/kg 297 mg/kg 1129/ Sample Acesulfame K Aspartame Rebaudioside A Saccharin Stevioside Sucralose Unit Diet cola mg/l Candy 145 mg/kg Diet tea 379 mg/l Marmalade 297 mg/kg Pudding mg/kg Table-top sweetener mg/kg Table-top sweetener mg/kg Standard Standard Table-top sweetener 2 Table-top sweetener A SIR ACQUITY UPLC H-Class ACQUITY QDa 17
18 ACQUITY UPLC H-Class 1. D J Yang, B Chen. Simultaneous determination of non-nutritive sweeteners in foods by HPLC/ESI-MS. J Agric Food Chem. 57(8): , ACQUITY QDa ACQUITY QDa UV UV 1 UV MS TEL FAX F TEL FAX Waters ACQUITY UPLC UPLC MassLynx QDa The Science of What s Possible Waters Corporation 2014 Waters Corporation. Printed in Japan JA PDF 18
19 ACQUITY UPLC H-Class /ACQUITY QDa Mark E. Benvenuti, Dimple Shah, and Jennifer A. Burgess Waters Corporation, Milford, MA, USA ACQUITY QDa ACQUITY QDa RI ELS ACQUITY UPLC H-Class ACQUITY QDa ACQUITY UPLC BEH Amide Sep-Pak C 18 UV HPLC 1,2 RI ELS RI RI ELS ESI MS Waters ACQUITY QDa ACQUITY QDa PDA LC MS QDa HPLC MS ACQUITY QDa ACQUITY UPLC H-Class QDa myo - 19
20 UPLC ACQUITY UPLC H-Class ACQUITY UPLC BEH Amide 1.7 µm mm myo mg/l mg/l / 10 mm NH 4 HCO 3 /0.1%NH 4 OH ml / 0.7 µl Chavaz-Servin 3 Sep-Pak C 18 Chavaz-Servin MS MS ACQUITY QDa ESI 0.8 V 4.0 V Hz m/z SIR m/z 1 1 SIR (m/z) [M+Cl - ] [M+Cl - ] [M-H + ] [M+Cl - ] [M+Cl - ] [M-H + ] -. myo- SIR myo- m/z myo- 180 ESI myo- m/z 179 m/z 215 [M+Cl-]- [M-H+]- MS 4 myo- m/z179 m/z 341 m/ z 377 m/z ppm SIR 1 [M-H+]- [M-H+]- 20 ACQUITY UPLC H-ClassACQUITY QDa
21 Fructose [M+Cl - ] - m/z Sucrose [M-H + ] - m/z Myo-inositol [M-H + ] - m/z ppm SIR m/ z 179, 215, Sucrose [M+Cl - ] - m/z 377 Glucose [M+Cl - ] - m/z Myo-inositol [M+Cl - ] - m/z Maltose [M+Cl - ] - m/z Lactose [M+Cl - ] - m/z Fructose [M-H + ] - m/z Glucose [M-H + ] - m/z Maltose [M-H + ] - m/z Lactose [M-H + ] - m/z Minutes 2A. m/z 377 2B. m/z 179 myo- myo myo- RI ELS MS myo- m/z 2 myo- SIR m/z 377, m/z A. m/z 377 2B. myo- m/z 179 MS m/z myo- LC MS 3 2 ACQUITY UPLC H-Class / ACQUITY QDa 21
22 3. m/z 377 A. 20 ppm B. C UV ACQUITY UPLC H-Class ACQUITY QDa UPLC MS RI ELS MS UV m/z ACQUITY QDa 1 1. L C Nogueiraa, F Silvab, I M P L V O Ferreirab, L C Trugoa. Separation and quantification of beer carbohydrates by high-performance liquid chromatography with evaporative light scattering detection. J Chrom A (2): , February I M P L V O Ferreira, A M P Gomes, M A Ferreira. Determination of sugars, and some other compounds in infant formulae, follow-up milks and human milk by HPLC-UV/RI. Carbohydr Polym. 37 (3): , J L Chávez-Servín, A I Castellote, M C López-Sabater. Analysis of mono- and disaccharides in milk-based formulae by high-performance liquid chromatography with refractive index detection. J Chrom A (2): , E Rogatsky, H Jayatillake, G Goswami, V Tomuta, D Stein. Sensitive LC-MS Quantitative Analysis of Carbohydrates by Cs+ Attachment. JASMS. 16 (11): , H E Indyk, D C Woollard. Determination of free myo-inositol in milk and infant formula by high-performance liquid chromatography. Analyst. 119: , M Kizito, E Iheanacho. Comparative Studies of the Nutritional Composition of Soy Bean (glycine max) and Lima Bean (phaseolus lunatus). Scientia Africana. 9 (2): 29-35, TEL FAX F TEL FAX Waters ACQUITY UPLC UPLC Sep-Pak QDa The Science of What s Possible Waters Corporation 2014 Waters Corporation. Printed in Japan JA PDF 22
23 ACQUITY QDa Jinchuan Yang, Rich DeMuro, and Joe Romano Waters Corporation, Milford, MA, USA ACQUITY QDa UV/Vis PDA ACQUITY QDa 1 2 PMF 2 3, 4 1 ACQUITY QDa UV/Vis ACQUITY UPLC H-Class ACQUITY UPLC PDA ACQUITY QDa MassLynx v. 4.1 MS ACQUITY QDa 1. 23
24 DMSO / LC LC ACQUITY UPLC H-Class ACQUITY QDa MS ESI+ SIR ACQUITY UPLC HSS T3 1.8 µm mm µl 5 V kv 5 Hz A B 0.6 ml/ 0.1% 0.1% 13 MassLynx 4.1 MS SCN 888, Build 274 A B ( ) (%) (%) ACQUITY QDa
25 UV/Vis UV/Vis PDA PDA UV/Vis PDA PDA MSPDA QC ACQUITY UPLC H-Class ACQUITY QDa ACQUITY QDa UV SIR 1 m/z m/z m/z 2 UV 325 nm ACQUITY QDa 2 E H UV/Vis PDA ACQUITY QDa 2 m/z m/z m/z (Da) ACQUITY QDa 25
26 C UV (325 nm) 3.0e-2 AU 2.0e-2 1.0e A B D E F G Time H 90 ACQUITY QDa SIR E F G H C % A B D -10 Time UV SIR A B CDE F G H ACQUITY QDa UV/Vis 1/ QC 2. n =10 % RSD % ACQUITY QDa
27 ACQUITY QDa Waters ACQUITY UPLC H-Class ACQUITY QDa 1. HS Lee, BW Widmer. Phenolic compounds. In: LML Nollet, ed. Handbook of Food Analysis. New York: Marcel Dekker, 1996, G Gattuso, D Barreca, C Gargiulli, U Leuzzi and C Caristi. Flavonoid composition of citrus juices. Molecules 2007, 12, AOAC Official Method Naringin and Neohesperidin in Orange Juice, Official Method of Analysis, AOAC. 4. W Widmer. Determination of Naringin and Neohesperidin in orange juice by liquid chromatography with UV detection to detect the presence of grapefruit juice: collaborative Study. J. AOAC Int. 83(5): 1155, TEL FAX F TEL FAX Waters ACQUITY UPLC MassLynx The Science of W hat s Possible Waters Corporation ACQUITY QDa Waters Corporation 2016 Waters Corporation. Produced in Japan JA PDF 27
28 Myco6in1 + ACQUITY QDa 12 Dominic Roberts, 1 Sara Stead, 1 Veronica Lattanzio, 2 Biancamaria Ciasca, 2 Eimear McCall, 1 Jennifer Burgess, 3 and Stephen Powers 4 1 Waters Corporation, Wilmslow, UK; 2 National Research Council of Italy, Institute of Sciences of Food Production, Bari, Italy 3 Waters Corporation, Milford, MA, USA; 4 VICAM, Milford, MA, USA 12 in vivo EU DONB1 B2 B1 B2 G1 G2 A 1881/ /2007 T2 HT / /2006 DON NIV CORTECS UPLC C 18 ACQUITY UPLC I-Class ACQUITY QDa VICAM Myco6in1 IAC LC-MS QDa 28
29 UPLC LC ACQUITY UPLC I-Class A B 12 CORTECS C µm mm 0.1% 2 mm 0.1% 2 mm 0.4 ml/ 10 µl A B (min) (%) (%) ml 60 ml 5 ml 2 ml 5mL VICAM Myco6in1+ IAC 10 ml 2mL 3 ml 10:90 v/v = : A 0.2 ml Myco6in1 + IAC :90 v/v = : A MS MS ACQUITY QDa ESI kv 5 Hz SIR 1 Myco6in1+ ACQUITY QDa 12 29
30 Waters ACQUITY QDa SIR m/z V µg/kg 1. NIV [NIV-H 2 O+H + ] to DON [DON+H + ] to G2 AFG2 [AFG2+H + ] to G1 AFG1 [AFG1+H + ] to B2 AFB2 [AFB2+H + ] to B1 AFB1 [AFB1+H + ] to HT2 HT2 [HT2+Na + ] to B1 FB1 [FB1+H + ] to T2 T-2 [T-2+NH 4+ ] to A OTA [OTA+H + ] to Zear [ZEA-H] to B2 FB2 [FB2+H + ] to AFG1 DON 1 DON µg/kg AFG µg/kg Compound name: AFG1 Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None A Compound name: DON Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None B 5.0 Residual Conc Residual Conc Response Response Conc -0 Conc A G µg/kg µg/kg 30 Myco6in1+ ACQUITY QDa 12
31 2 - S/N 4 8 MS LC 8. FB1 9. T-2 S/N NIV DON AFG NIV 10. OTA AFG AFB AFB DON 3.AFG2 4.AFG1 5.AFB2 7. HT-2 6.AFB1 11. Zear 12. FB2 HT FB T OTA Zear FB EU S/ N 12 ACQUITY QDa TEL FAX F TEL FAX Waters UPLC, ACQUITY UPLC The Science of W hat s Possible Waters Corporation 2016 Waters Corporation. Produced in Japan JA PDF 31
32 ACQUITY UPLC H-Class ACQUITY QDa Brian Tyler and Jennifer A. Burgess Waters Corporation, Milford, MA, USA ACQUITY QDa UPLC HPLC SIR Selected Ion Recording RADAR ACQUITY QDa 1 FDA maximum residue level, MRL 50 µg/kg 1 EU 25 µg/kg 10 µg/kg 2 LC UV 273 nm 1 5- HMF ACQUITY QDa ACQUITY UPLC H-Class MassLynx TargetLynx Oasis HLB ACQUITY QDa
33 UPLC LC ACQUITY UPLC H-Class ACQUITY UPLC BEH Shield RP mm 1.7 µm 40 MS MS ACQUITY QDa ESI 0.8 kv µl 150 A B 0.60 ml/ 0.1% 0.1% 50:50 2 V SIR HMF SIR Hz 3 50:50 90:10 Time Flow rate %A %B Curve (min) (ml/min) Initial UPLC Oasis HLB 3 cc/60 mg 3 ml 3 ml 2.5 ml 1 1% 1 g/100 ml 3 ml 2 0.1% 1 ml : -t- MTBE 10: ml 500 µl µg/l µg/l ACQUITY UPLC H-Class ACQUITY QDa 33
34 UV HMF MS 3 MS MS ACQUITY QDa ACQUITY QDa MS ACQUITY UPLC ACQUITY QDa LC 5- HMF SIR Single ion recording RADAR UV 273 nm UV 2 2 m/z 153 HMF m/z 125 MS UV MS HMF UV MS 3 2 SIR MS 1 µg/l S/N 10 5 µg/l EU FDA 1/10 EU 1/2 A. 1 µg/l patulin B. 5 µg/l patulin Peak-to-peak S/N = 4 Peak-to-peak S/N = SIR A.1 µg/l B. 5 µg/l S/N 34 ACQUITY UPLC H-Class ACQUITY QDa
35 µg/l r µg / L RADAR % ACQUITY UPLC H-Class ACQUITY QDa 35
36 µg/l MS SIR UV m/z RADAR SIR ESI BPI base peak intensity HMF SIR 5 A. BPI B. SIR C. HMF SIR 5. A BPI base peak intensity B50 µg/l SIR C HMF SIR RADAR 36 ACQUITY UPLC H-Class ACQUITY QDa
37 5 µg/l 50 TargetLynx Trend Plot 6 %RSD 9% EU 2002/657/EC µg/l µg/l ACQUITY UPLC H-Class ACQUITY QDa 1/10 EU 1/2 RADAR SIR 1. US FDA website: NaturalToxins/ucm htm 2. Commission Regulation (EC) No. 1881/2006 Section J Morphet, Rapid Analysis of Patulin Contaminination in Apple Juice. Waters Application Note no en, Commission Directive 2002/657/EC, Off. J. of the European Communities No. L221/8. ACQUITY QDa MS MS TEL FAX F TEL FAX Waters ACQUITY ACQUITY UPLC UPLC MassLynx Oasis The Science of What s Possible Waters Corporation RADAR TargetLynx QDa Waters Corporation 2013 Waters Corporation. Printed in Japan JA PDF 37
38 ACQUITY UPLC I-Class/ACQUITY QDa Eimear McCall, 1 Sara Stead, 1 Dominic Roberts, 1 and Jennifer A. Burgess 2 1 Waters Corporation, Manchester, UK 2 Waters Corporation, Milford, MA, USA LC MS Oasis HLB EU EU Commission Decision 2003/181/ EC MRPL 0.3 µg/ kg ELISA ELISA 16% 1 MS LC ACQUITY QDa LC ACQUITY UPLC HSS C 18 ACQUITY UPLC I-Class ACQUITY QDa ACQUITY QDa 38
39 UPLC LC ACQUITY UPLC I-Class 5 : ACQUITY UPLC HSS C µm mm 10 mm / µl ACQUITY QDa ESI - SIR V 600 / kv 5 / 5 g 25 ml 0.3 µg/kg 5 ml 70% 15 ml 3,900 G ml 10% 15 ml Oasis HLB 3 cc 60 mg 2 ml 4 ml 15 ml 5% 6 ml 100% 6 ml 10% 300 µl LC-MS µg/kg 6 MS MS 15 V EU MRPL µg/kg R 2 >0.995 Compound name: CAP Correlation coefficient: r = , r 2 = Calibration curve: * x Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: Null, Axis trans: None 20.0 Residual ng/ml Response ng/ml µg/kg 2 ACQUITY UPLC I-Class/ACQUITY QDa 39
40 2 Slope sample ( x 100 ) Slope standard 177% n=2 3A EU MRPL n=4 1 2 n=8 3B n=8 S/N B. MM milk y = x R² = A. Blank milk sample Peak area A. Solvent y = x R² = Concentra on (ppb) B. Chloramphenicol in milk at 0.3 g.kg µg/kg 2A10% 2B 3. Selected ion recording SIR m/z 321 3An=2 3B MRPL 0.3 µg/kg n=8 S/N ACQUITY UPLC I-Class/ACQUITY QDa
41 3B % EU Commission Decision 2002/657/EC n=8 RSD 4.6% 1. n=8 % %RSD V Gaudin, N Cadieu, O Maris O. IVth International Symposium on Hormone and Veterinary Drug Residue Analysis, Belgium, EU MRPL 0.3 µg/kg TEL FAX F TEL FAX Waters ACQUITY UPLC ACQUITY QDa The Science of What s Possible Waters Corporation 2014 Waters Corporation. Printed in Japan JA PDF 41
42 Alliance HPLC 2-4- Mark E. Benvenuti and Jennifer A. Burgess Waters Corporation, Milford, MA, USA HPLC-MS SPE CORTECS HILIC 4-4-MEI IARC 1 EC 250 mg/kg 2 Proposition MEI No Significant Risk Level: NSRL 29 µg/ PDA 4-MEI LC-MS 4 SPE 4 CORTECS HPLC 4-MEI 2-MEI Alliance HPLC ACQUITY QDa CORTECS QDa HILIC 42
43 LC HPLC Alliance 20.5 CORTECS HILIC µm mm A 10 mm ph4 B 0.1% 5 µl Time Flow %A %B 1. Initial MS ACQUITY QDa Performance ESI+ 0.8 kv 15 V Schlee mg/l 2-MEI 4-MEI 10 ppm 2-MEI 4-MEI 10 ppm 10 ppm ppb 100 µl 900 µl 2-4-MEI (ppb) (IS)(ppb) (MEI) SIR m/z SIR 2-4-MEI 83.1 m/z 2 Hz Alliance HPLC
44 1 m/z MEI m/z MEI 2 4-MEI R ppb % MEI 2-MEI MEI 2 4-MEI m/z MEI 2-MEI Intensity Imidazole (IS) Intensity m/z Minutes ppb Area Ratio R Amount 2. 4-MEI 44 Alliance HPLC 2-4-
45 Intensity Cola 1 4-MEI A Intensity Cola 2 4-MEI B Intensity Cola 3 4-MEI C Intensity Orange Flavored Soft Drink D Intensity Whiskey 1 E Intensity Whiskey 2 4-MEI 2-MEI F Minutes 3. 6 m/z83.1 SIR A-C: 1-3 D: E-F: 1 2 ; D-F 4-MEI (ppb) 2-MEI (ppb) 2. ppb 4-MEI spike (ppb) 2-MEI spike (ppb) %R 4-MEI (ppb) %R 2-MEI (ppb) Cola ND Cola ND Cola ND Orange SD ND ND Whiskey 1 ND ND Whiskey Alliance HPLC
46 4-MEI 1 kg 80 mgnoael EU MEI 110%-123% %RSD RT %RSD ppb %RSD 4-MEI MEI ppb 2-4-MEI % 7 %RSD 4-MEI 0.82% 2-MEI 1.02% 4-MEI 4-MEI 2-MEI 4-MEI 2-MEI Alliance HPLC ACQUITY QDa CORTECS HILIC 4-MEI MEI 4-MEI 1. IARC Working Group on the Evaluation of Carcinogenic Risks to Humans, IARC Monogr. Eval. Carcinog. Risks Hum., Monographs/vol101/mono pdf. 2. European Food Safety Authority (2004) Scientific opinion on the re-evaluation of caramel colours (E 150 a,b,c,d) as food additives. EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS). EFSA Journal 2011: 9 (3), pp.5 doi: /j.efsa Specific Regulatory Levels Posing No Significant Risk: 4-Methylimidazole (4-MEI). Proposition 65. OEHHA. 7 Oct Retrieved 15 Aug C Schlee, M Markovaa, J Schranka, F Laplagnea, R Schneidera, DW Lachenmeiera. Determination of 2-methylimidazole, 4-methylimidazole and 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole in caramel colours and cola using LC/MS/MS; J Chrom B. 27: , UV SIR TEL FAX F TEL FAX Waters CORTECS Alliance ACQUITY QDa The Science of W hat s Possible Waters Corporation 2015 Waters Corporation. Produced in Japan JA PDF 46
47 Mark E. Benvenuti, Michael S. Young, Gareth E. Cleland, and Jennifer A. Burgess Waters Corporation, Milford, MA, USA CA DCD 3 LC-MS/MS LC UV CA DCD 3 1 mg/kg 2.5 mg/kg TDI bw 1kg 0.64 mg 4 TDI 1kg 0.2 mg 5 DCD 1kg 1 mg 6 MS/MS HILIC 7 CA DCDWaters ACQUITY QDa ACQUITY UPLC H-Class ACQUITY UPLC H-Class ACQUITY QDa ACQUITY UPLC BEH Amide Sep-Pak MassLynx Empower 3 DCD CA DCD DCD CA 1. CA DCD 47
48 LC LC ACQUITY UPLC H-Class Empower ACQUITY UPLC BEH Amide 1.7 µm mm 35 A 10 mm 0.125% 50% B 10 mm 0.125% 90% 0.6 ml/ 5 µl : : : SIR : SIR : 0.8 kv 0.6 kv m/z V m/z V ml/ %A %B 1. Initial mg/l 2 mg/l 100 mg/l 100 mg/l 90% µg/l 1000 µg/l 1000 µg/l 1 90% 9 MS MS : ACQUITY QDa Performance : ESI+/- SIR m/z 85.1 : 10 V : 5 Hz : m/z 50 to 300 : 15 V 1 g 2% 10mL 1 ml 9 m L 2% 1 ml 9 ml g rcf 20 1mL 90% 6mL Sep-Pak 6 cc p/n % 4mL 5 1 g 1 mg/ L 20 mg/l
49 µg/l mg/l 20 mg/l 3 A 0.30 HILIC 2 AU 0.15 HILIC HILIC BEH HILIC BEH Amide A HILIC BEH ACQUITY UPLC BEH Amide 2B 2B B AU Amide A. ACQUITY UPLC BEH HILIC mm SIR 280 nm UV 2B. ACQUITY UPLC BEH Amide mm SIR 280 nm UV 49
50 3 1 UV * 20 µg/ L 1000 µg/ L SIR 3 UV SIR 3 AU UV nm m/z , m/z , 5 SIR MS SIR m/z104.1 Sep- Pak Sep-Pak 4 4A SIR 4B m/z B 4C SIR 4D m/z m/z , UV * SIR x10 7 2x Sep-Pak m/z m/z A SIR B m/z 104 C SIR D m/z 104 * UV 3 PDA 2 50
51 m/z /250 SIR SIR MS A R g/l 5A-C % A 1 mg/l 6B 6C 20 mg/l 5 ) 6A 2.5 B C g/l R g/l R A B C 51
52 RSD 3 3 A B SIR m/z SIR m/z m/z C SIR m/z 85.1 m/z mg/l 20 mg/l 20 mg/l SIR A SIR m/z B SIR m/z C SIR m/z
53 5 3 75% 123% ACQUITY QDa BEH Amide ACQUITY UPLC H-Class 3 1. Import alert #99 29, Detention without physical examination of all vegetable protein products from China for animal or human food use due to the presence of melamine and/or melamine analogs. U.S. Food and Drug Administration. 27 April Culprit in pet food deaths may be combination of contaminants. Michigan State University. November 29, Government downplays DCD risk. News NZ. January 27, US FDA, Interim Melamine and Analogues Safety/ Risk Assessment May 25, 2007, http// melamra.html 5. The EFSA Journal 2010, 8, The EFSA Journal (2004) 36, Draher J et al. Validation of a rapid method of analysis using ultrahigh-performance liquid chromatography tandem mass spectrometry for nitrogen-rich adulterants in nutritional food ingredients Journal of Chromatography A. 1373: ; TEL FAX F TEL FAX Waters ACQUITY UPLC ACQUITY QDa ACQUITY UPLC Sep-Pak MassLynx Empower Empower The Science of W hat s Possible Waters Corporation 2016 Waters Corporation. Produced in Japan JA PDF 53
54 NOTES 54
55 NOTES 55
56 Austria Australia Belgium and Luxembourg Brazil Canada China Czech Republic Denmark Finland France Germany Hong Kong Hungary India Ireland Israel Italy Japan Korea Malaysia Mexico The Netherlands Norway Poland Portugal Puerto Rico Singapore Spain Sweden Switzerland Taiwan UAE UK US TEL FAX F TEL FAX Waters ACQUITY ACQUITY UPLC Alliance CORTECS Empower MassLynx Oasis QDa Sep-Pak UPLC The Science of What s Possible Waters Corporation RADAR TargetLynx TrendPlot Waters Corporation VICAM Myco6in1 + VICAM A Waters Business 2016 Waters Corporation. Printed in Japan JA 03A H
1 WSV SIR m/z Analyte RT (Min) SIR m/z Cone voltage (V) Ascorbic Acid (C).91 177 2 Thiamine (B1) 1.1 265 5 Nicotinic Acid (B3) 1.27 124 15 Pyridoxal (
ACQUITY UPLC H-Class ACQUITY QDa Mark E. Benvenuti, Dimple Shah, and Jennifer A. Burgess Waters Corporation, Milford, MA, USA MS ACQUITY QDa UPLC HPLC UV ACQUITY QDa LC EC 1925/26 CFR 21 HPLC 1 LC-MS ACQUITY
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ACQUITY UPLC I-Class/Xevo TQD Mark Roberts and Michelle Wood Waters Corporation, MS Technologies Centre, Manchester, UK ACQUITY UPLC I-Class/Xevo TQD 2 2 MRM 178 / 950 MS ACQUITY UPLC/ACQUITY TQD 2 1-3
Waters Sigma-Aldrich 1.0 mg/ml LC Waters Alliance HPLC ACQUITY UPLC H-Class Bio 30 cm Alliance HPLC UV ACQUITY UPLC TUV mm nm XBridge Protein B
HPLC 3. µm BEH Stephan Koza, Susan Serpa, Hua Yang, Edouard Bouvier, and Kenneth J. Fountain Waters Corporation, Milford, MA, USA HPLC 200 0 HPLC 2 2010 UPLC 200 SEC 1 UPLC SE-UPLC 2 µm BEH BEH 100% SE-UPLC
LC LC ACQUITY UPLC H-Class ACQUITY UPLC CSH C mm 1.7 µm : THF/ 2/8 A 1 mm / 4/6 THF 2 mg/ml THF/ 2 THF 2 ACQUITY UPLC CSH C 18
ACQUITY UPLC H-Class/QDa Nihon Waters K.K., Tokyo, Japan TAG 1 3 1 1 8 9 TAG TAG TAG TAG TAG UV UV APCI ESI MS TAG Sn-2 Sn-3 Sn-1 C18: ACQUITY UPLC CSH C 18 ACQUITY UPLC H-Class 1. ACQUITY QDa TAG ACQUITY
平成26年度 化学物質分析法開発報告書
2,2-2,2 -Iminodiethanol Diethanol amine CAS 111-42-2 C 4 H 11 NO 2 105.1356-105.0790-28 C 1) 1.0881 g/cm 3 1)
HILIC UPLC/MS UPLC LC/ MS LC/MS HPLC 2-4 HPLC 4,5 HILIC / HILIC 80 ACQUITY UPLC Xevo QTof MS ACQUITY BEH HILIC HILIC TOF ESI 1.7 µm BEH HILIC UPLC HIL
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平成26年度 化学物質分析法開発報告書
2,2-2,2 -Iminodiethanol Diethanol amine CAS 111-42-2 C 4 H 11 NO 2 105.1356-105.0790-28 C 1) 1.0881 g/cm 3 1)
LC ACQUITY UPLC H-Class Bio TUV Sigma, mg/ml 0.01 N 276 nm μl 0.4 ml/min L g/l / 99% / 65/15/20 v/v/v 10% 10% 0.01 N ACQUITY
Ultra Performance Liquid Chromatography Stephan Koza, Paula Hong, and Kenneth J. Fountain Waters Corporation, 34 Maple Street, Milford, MA, USA & SEC 2010 2 8 5 2030 4 3 8 1982 2009 131 USP EPHMW USP34
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ACQUITY UPC 2 ACQUITY UPC 2 A A+D3 D ml E 10 ml K μm K μm PDA ACQUITY UPC 2 BEH mm, 1.7 μm ACQUITY UPC 2 HSS C 18 SB 3.
UPC 2 Andrew Aubin Waters Corporation, Milford, MA, USA Waters ACQUITY UPC 2TM 6 tert- UltraPerformance Convergence Chromatography TM UPC 2 ACQUITY UPC 2 1 A A D3 E D3 K1 K2 UPC 2 A E K1 ACQUITY UPC 2
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[ cortecs columns ] 可能性の限界に挑戦するµm 以下の ソリッドコアパーティクルカラム C8 C8+ HILIC 究極のカラム効率と 性能を実現する ソリッドコアパーティクル カラム µm 以下のパーティクルの性能を最大 限に発揮させるための独自の研究成果に より ウォーターズは粒子径 µm 以下 の UltraPerformance LC カラムファミリー に 新製品を追加しました.6
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1 Agilent 12 Infinity 1 Siji Joseph Agilent Technologies, Inc. Bangalore, India 12 1 2 4 5 3 2 1 4 1 12 14 4 2 Agilent 12 Infinity 2 4 5 3 2.5 1. 1.5 2. 2.5 3. 2 (RP-HPLC) Agilent Poroshell EC-C1 Agilent
平成26年度 化学物質分析法開発報告書
N,N- N,N-Dimethylacetamide Acethyldimethylamine CAS 127-19-5 C 4 H 9 NO 87.12 ~ 87.14 87.68413 163-165 C 1) - 18.59 C 2) 1 mg/l 25 C 3) 3.3 hpa 2 C 4) log P ow -.77 2).9429 2/4 C 1) 3.1 4) 1.31E - 8 atm-m
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3--2-3-Iodo-2-propynyl butylcarbamate (IPBC) Carbamic acid, butyl-, 3-iodo-2-propynyl ester Iodocarb CAS 55406-53-6 C 8 H 12 NO 2 I C (g/cm 3 ) (mmhg) log P ow 281.09 (280.9910) 64 68 1.51 1.57 (20 C)
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2005 10 13 Dr. RGA 19602000 10 Australian Institute of of Health and and Welfare 2004 ABS Cat. Cat. No. No. 3201.0; AIHW National Population Database 1875 2 211997 8 122164 http://www.wowzone.com/calment.htm
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1 2 3 4 Alteration of program parameters Effect Increase hybridization temperature Can prevent false positive signals, decrease background, can also increase dynamic range Decrease hybridization temperature
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Hydrocortisone Sodium Phosphate 3.5 1.0 0.5 1 μg/ml 3.0 2.5 25 ng/ml 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 min 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 min Fig.1 Chromatograms of Hydrocortisone
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Tl Thallium CAS 7440-28-0 CH 3 COOTl Thallium Acetate CAS 563-68-8 Tl 2 CO 3 Thallium Carbonate CAS 6533-73-9 Tl 2 SO 4 Thallium Sulfate CAS 7446-18-6 TlNO 3 Thallium Nitrate CAS 10102-45-1 TlClO 3 Thallium
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1 9924 2260 CAS 107186 H 2 C CH CH 2 OH 2- -1- C 3 H 6 O 58.08 1 99 1. 2, 3 3, 4, 5-129 96-97 2 21c.c. 24o.c. 2 6, 7 378 6, 7 2.5-18 d 20 4 0.8540 2, 5 2.00 = 1 2.7 kpa 20 mmhg204.3 kpa 32 mmhg30 4 log
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EC declaration of conformity We, Grundfos, declare under our sole responsibility that the products SPK, CRK and MTR, to which this declaration relates
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