Method for simultaneously detecting five anti-tuberculosis drugs in blood plasma via UPLC-MS/MS method
An anti-tuberculosis and plasma technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that cannot fully meet the needs of clinical measurement, long analysis time, complicated pre-treatment, etc., and achieve short processing time, short analysis time, and pre-processing Handle simple effects
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Embodiment 1
[0054] Different chromatographic columns, mobile phase ratios and gradient elution methods were used to analyze the detected standard substances.
[0055] (1) The chromatographic column Shim-pack XR-ODSⅢ (2.0mm i.d×50mm, 1.6μm); the pre-column Shim-pack GIST-HP(G) (2μm C18, 2.1×10mm) was used to separate and analyze the standard. The mobile phase AB adopts different proportions and gradient elution methods. The peaks of isoniazid and ethambutol in the analyte both elute at about 0.5 min, and the above two analytes are not retained on this chromatographic column. Wherein the chromatographic mobile phase is set as follows: A (water phase): 0.01% (w / w) formic acid aqueous solution, B (organic phase): acetonitrile solution with formic acid concentration of 0.01% (w / w).
[0056]
[0057] The chromatographic peaks of the five standard substances measured are as follows: figure 1 ,Depend on figure 1 It can be seen that ethambutol and isoniazid are almost not retained, and the pe...
Embodiment 2
[0076] Three plasma samples from patients infected with Mycobacterium tuberculosis were collected respectively, named sample 1, sample 2, and sample 3.
[0077] (1) a. Standard product handling:
[0078] Take 90 μL of blank plasma, add 10 μL of mixed standard solutions of different concentrations, vortex and mix, add 10 μL of mixed isotope internal standard working solution, then add 300 μL of acetonitrile, vortex for 1 min, and centrifuge at 12000 r / min for 5 min; absorb 300 μL of supernatant in another Add 300 μL of ultrapure water to a clean 1.5 mL centrifuge tube, vortex for 30 seconds, and filter through a 0.45 μm filter to obtain the test solution; take 2 μL of the supernatant for analysis, and record the chromatogram;
[0079] The quantitative serial concentrations of rifampicin are: 0.039, 0.078, 0.156, 0.313, 0.625, 1.25, 2.5, 5μg / mL,
[0080] The quantitative serial concentrations of rifabutin are: 0.016, 0.031, 0.063, 0.125, 0.25, 0.5, 1, 2μg / mL,
[0081] The quan...
Embodiment 3
[0091] (1) Matrix effect detection
[0092] Take 90 μL of blank plasma, add 10 μL of mixed standard solutions of different concentrations, vortex and mix, add 10 μL of mixed isotope internal standard working solution, then add 300 μL of acetonitrile, vortex for 1 min, and centrifuge at 12000 r / min for 5 min; absorb 300 μL of supernatant in another Add 300 μL of ultrapure water to a clean 1.5 mL centrifuge tube, vortex for 30 seconds, and filter through a 0.45 μm filter to obtain the test solution; prepare 6 parallel samples, and take 2 μL of the supernatant for analysis.
[0093] Use 40% acetonitrile aqueous solution to prepare low and high working solutions of two concentrations (i.e. the concentrations corresponding to the compounds in the quality control sample in the summary of the invention), and prepare 6 copies in parallel, and take 2 μL for analysis. The results are shown in Table 3.
[0094] Table 3: Matrix Effects of Protein Precipitation Experimental Methods
[009...
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