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Experimental Details
Sample Preparation
(1) Put Cleanert® PPT and 96-well plate onto the SPE manifold
(2) Add 50 μL of 20 ng/mL hydrochlorothiazide into each well
(3) Quickly add 250 μL ACN and stand for 3 min
(4) Suction by vacuum and dry with N2, then resolve with 200 μL mobile phase.
Instrumentation
Instrumentation: LC-MS/MS, API 4000
Column: Venusil® ASB C18, 2.1×150 mm, 5 μm;
Column temperature: 25℃
Mobile phase: 90% ACN, 10% 0.01 mol/L ammonium acetate aq (0.1% formic contained)
Flow rate: 0.2 mL/min;
Sample injection: 5 μL;
Ion source: ESI - Negative
Scan mode: MRM
Compound |
Parent ion |
Daughter ion |
Hydrochlorothiazide |
295.7 |
269.0 |
Table 1 MS/MS transitions information of Hydrochlorothiazide
Results
Figure 2 Chromatogram of Hydrochlorothiazide standard solution
Figure 3 Chromatogram of plasma sample spiked with Hydrochlorothiazide
Concentration (ng/mL) |
Recovery (%) |
Average recovery (%) |
RSD (%) |
|||||
1 |
2 |
3 |
4 |
5 |
6 |
|||
20 |
79.4 |
75.8 |
76.5 |
74.9 |
75.1 |
78.4 |
76.7 |
2.39 |
Table 2 Recovery data of spiked sample
Ordering Information
Products |
Specification |
Cat.No |
Cleanert® PPT |
2 mL/well |
96CD2025-Q |
Venusil® ASB C18 |
2.1×150 mm, 5 μm, 150 Å |
VS951502-0 |
96-well collection plate |
2.2 mL Squaral well |
96SP2036-2 |
96-well vacuum manifold |
adapt to 96 well plate |
VM96 |
NV-96G for 96 Well Plates |
adapt to 96 well plate |
NV96-G |
Acetonitrile |
HPLC, 4 L |
AH015-4 |
1.5 mL vials |
1.5 mL short thread vial, amber glass, label and filling lines |
AV1111-0 |
1.5 mL vials caps |
9 mm screw neck cap, center hole; red silicone/ white PTFE septa |
AV2100-0 |
Micro-insert clear glass |
300 μL micro-insert, 31×6 mm |
AV1132-6 |