RESUMO
OBJECTIVES: To establish a rapid method for the analysis of bucinnazine in blood by UPLC-MS/MS and to apply the method to the practical case. METHODS: After the internal standard was added to blood, the protein was precipitated with 900 µL mixed solution (Vacetonitrileâ¶Vwater=8â¶2). After vortex and centrifugation, the protein was measured through 0.22 µm filter membrane. The separation was performed on C18 chromatography column, with acetonitrile and 5 mmol/L ammonium acetate containing 0.1% formic acid aqueous as mobile phase gradient elution at the flow rate of 0.4 mL/min. Multiple reaction monitoring scan was performed in electrospray positive ion mode, quantitative measurement was performed by internal standard method, and methodological verification was carried out. RESULTS: The linear relationship of bucinnazine in blood was good in the range of 0.5-200 µg/L, the correlation coefficient (r) was 0.999 7, the limit of detection was 0.1 µg/L, the limit of quantitation was 0.5 µg/L, and the recovery was 78.3%-83.8% at 1, 10 and 100 µg/L mass concentration levels. The matrix effect was 69.4%-73.8%, the intra-day precision was 1.9%-2.8%, and the inter-day precision was 2.8%-3.2%, the accuracy was 3.1%-3.5%. The stability test results of 1 and 100 µg/L mass concentrations at -25 â showed that the accuracy (bias) of 10 d was less than 4.5%. CONCLUSIONS: This method has the advantages of simple pre-treatment process, fast sample processing speed, high sensitivity of instrument analysis, good stability of content determination and reliable identification results, and can meet the needs of case identification.
