Department of Clinical Pharmacology and Pharmacokinetics, Ranbaxy Research Laboratories, Plot: GP-5, Sec-18, HSIIDC, Old Delhi-Gurgaon Road, Gurgaon-122 015, Haryana, India.
J Pharm Biomed Anal. 2009 Dec 5;50(5):966-76. doi: 10.1016/j.jpba.2009.06.050. Epub 2009 Jul 7.
A very sensitive, reliable, reproducible and highly selective assay for the simultaneous determination of free and total (conjugated and unconjugated) propranolol and its equipotent hydroxyl metabolite, 4-hydroxy propranolol, in human plasma was developed and validated. The analytes were simultaneously extracted from 0.300 mL of human plasma using solid phase extraction and detected in positive ion mode by tandem mass spectrometry with a turbo ionspray interface. Deuterium-labeled propranolol and 4-hydroxy propranolol, propranolol-d7 and 4-hydroxy propranolol-d7, were used as internal standards. The method has a lower limit of quantitation (LOQ) of 0.20 ng/mL for both analytes with the limits of detection (LOD) 50 and 100 pg/mL for propranolol and 4-hydroxy propranolol, respectively, based on a signal-to-noise ratio of 5. The assay was linear over a range 0.20-135.00 ng/mL for free propranolol and 0.20-25.00 ng/mL for free 4-hydroxy propranolol and linear over range 1.00-500.00 ng/mL for total propranolol and 1.00-360.00 ng/mL for total 4-hydroxy propranolol, with coefficient of determination greater than 0.99 for both analytes. The extraction recoveries were >96 and >64% on an average for propranolol and 4-hydroxy propranolol, respectively. The analytes were found stable in human plasma through five freeze (-15 degrees C)-thaw (room temperature) cycles and under storage on bench-top for at least 6.5 h, and also in mobile phase at 10 degrees C for at least 48 h. The method has shown tremendous reproducibility, with intra- and inter-day precision <11.3% (RSD), and intra- and inter-day accuracy <11% of nominal values, for both analytes, and has proved to be highly reliable for the analysis of clinical samples.
建立并验证了一种灵敏、可靠、重现性好且高度专属性的同时测定人血浆中游离（结合和非结合）和总（结合和非结合）普罗帕酮及其等效力代谢物 4-羟基普罗帕酮的方法。分析物用人血浆 0.300 mL 以固相萃取同时提取，正离子模式下串联质谱仪，涡轮离子喷雾接口检测。氘代普罗帕酮和 4-羟基普罗帕酮、普罗帕酮-d7 和 4-羟基普罗帕酮-d7 作为内标。该方法对两种分析物的定量下限（LOQ）均为 0.20ng/mL，检测限（LOD）分别为普罗帕酮和 4-羟基普罗帕酮的 50 和 100pg/mL，信号噪声比为 5。游离普罗帕酮和游离 4-羟基普罗帕酮的线性范围分别为 0.20-135.00ng/mL 和 0.20-25.00ng/mL，总普罗帕酮和总 4-羟基普罗帕酮的线性范围分别为 1.00-500.00ng/mL 和 1.00-360.00ng/mL，两种分析物的相关系数均大于 0.99。普罗帕酮和 4-羟基普罗帕酮的平均提取回收率分别为>96%和>64%。该方法经 5 次冻融（-15°C）循环和至少 6.5 h 室温放置后，在人血浆中稳定，在室温下，在乙腈流动相中至少 48 h 稳定。该方法具有良好的重现性，两种分析物的日内和日间精密度（RSD）均<11.3%，日内和日间准确度均<11%的名义值，该方法可用于临床样品的分析。
