Gabani Bhavesh Babulal, Sulochana Suresh P, Kiran Vinay, Todmal Umesh, Mullangi Ramesh
Drug Metabolism and Pharmacokinetics, Jubilant Biosys, Bangalore, India.
Biomed Chromatogr. 2019 Nov;33(11):e4661. doi: 10.1002/bmc.4661. Epub 2019 Sep 1.
A liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantification of tunicamycin in rat plasma as per regulatory guideline. Chromatography of tunicamycin and the IS in the processed plasma samples was achieved on an X-Terra phenyl column using a binary gradient (mobile phase A, acetonitrile and mobile phase B, 5 mm ammonium formate) elution at a flow rate of 0.6 ml/min. LC-MS/MS was operated under the multiple reaction monitoring mode using the electrospray ionization technique in positive ion mode and the transitions of m/z 817.18 → 596.10, 831.43 → 610.10, 845.29 → 624.10, 859.23 → 638.10 and 309.24 → 163.20 were used to quantitate homologs A-D and the IS, respectively. The total chromatographic run time was 4.5 min. The correlation coefficient (r ) was >0.99 for all homologs with accuracy 90.7-107.4% and precision 0.74-15.1%. The recovery of homologs was 78.6-90.2%. No carryover was observed and the matrix effect was minimal. Tunicamycin four homologs were found to be stable on the bench-top for 6 h, for up to three freeze-thaw cycles, in the injector for 24 h and for 1 month at -80°C. The applicability of the validated method has been demonstrated in a rat pharmacokinetic study.
已根据监管指南开发并验证了一种液相色谱-串联质谱（LC-MS/MS）方法，用于定量大鼠血浆中的衣霉素。在处理后的血浆样品中，衣霉素和内标的色谱分离是在X-Terra苯基柱上进行的，使用二元梯度（流动相A为乙腈，流动相B为5 mM甲酸铵）洗脱，流速为0.6 ml/min。LC-MS/MS在多反应监测模式下运行，采用电喷雾电离技术，正离子模式，m/z 817.18→596.10、831.43→610.10、845.29→624.10、859.23→638.10和309.24→163.20的跃迁分别用于定量同系物A-D和内标。总色谱运行时间为4.5分钟。所有同系物的相关系数（r）均>0.99，准确度为90.7-107.4%，精密度为0.74-15.1%。同系物的回收率为78.6-90.2%。未观察到残留，基质效应最小。发现衣霉素的四个同系物在台式上6小时内、最多三个冻融循环中、进样器中24小时内以及-80°C下1个月内均稳定。已在大鼠药代动力学研究中证明了该验证方法的适用性。
