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目的建立测定昆明小鼠血浆中RX31肽的在线固相萃取-液质联用法(on-line SPE HPLC-MS/MS)。方法血浆样品经乙腈沉淀蛋白后,取上清经on-line SPE,以甲醇和0.1%甲酸为流动相,资生堂CAPCELL PAK MGⅢ柱分析;采用电喷雾电离源,以多反应监测(MRM)方式进行正离子检测,采用外标法对RX31肽样品进行定量分析。结果与结论定量分析的离子反应为m/z 640.3→m/z 303.3。测定昆明小鼠血浆中RX31肽的线性范围为10～500 ng/ml,最低定量限(LLOQ)为10.0 ng/ml,方法学符合药代动力学研究要求。将该法应用于昆明小鼠单次皮下注射RX31肽(10、30和90 mg/kg)后的药代动力学研究中,利用单一的固相萃取柱完成了全部血浆样品的定量分析。此方法为RX31肽的非临床药代动力学研究提供了可靠的数据,对其临床研究具有重要的参考价值。
OBJECTIVE To establish an on-line SPE HPLC-MS / MS method for the determination of RX31 peptide in the plasma of Kunming mice. Methods The plasma samples were precipitated with acetonitrile and the supernatant was collected on on-line SPE using methanol and 0.1% formic acid as the mobile phase and analyzed by a Shiseido CAPCELL PAK MGIII column. Electrospray ionization (ESI) was used to conduct MRM (Multiple Reaction Monitoring) Positive ion detection, external standard method for quantitative analysis of RX31 peptide samples. RESULTS AND CONCLUSIONS The ion reaction for quantitative analysis was m / z 640.3 → m / z 303.3. The linear range of RX31 peptide in plasma of Kunming mice was 10-500 ng / ml and the lowest limit of quantification (LLOQ) was 10.0 ng / ml. The method was in accordance with the requirements of pharmacokinetic study. This method was applied to the pharmacokinetic studies after a single subcutaneous injection of RX31 peptide (10, 30, and 90 mg / kg) in Kunming mice using a single SPE column to perform a quantitative analysis of all plasma samples. This method provides reliable data for the non-clinical pharmacokinetic study of RX31 peptide and has important reference value for clinical research.