Department of Pharmaceutical Analysis, College of Pharmacy, Chung-Ang University, 221 Heukseok-Dong, Dongjak-Gu, Seoul 156-756, South Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Aug 15;878(24):2235-42. doi: 10.1016/j.jchromb.2010.06.031. Epub 2010 Jul 1.
A rapid and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry method (LC-ESI-MS/MS) was developed and validated for the determination of goserelin in rabbit plasma. Various parameters affecting plasma sample preparation, LC separation, and MS/MS detection were investigated, and optimized conditions were identified. Acidified plasma samples were applied to Oasis((R)) HLB solid-phase extraction (SPE) cartridges. Extracted samples were evaporated under a stream of nitrogen and then reconstituted with 100microL mobile phase A. The separation was achieved on a Capcell-Pak C18 (2.0mmx150mm, 5microm, AQ type) column with a gradient elution of solvent A (0.05% acetic acid in deionized water/acetonitrile=85/15; v/v) and solvent B (acetonitrile) at a flow rate of 250microL/min. The LC-MS/MS system was equipped with an electrospray ion source operating in positive ion mode. Multiple-reaction monitoring (MRM) of the precursor-product ion transitions consisted of m/z 635.7-->m/z 607.5 for goserelin and m/z 424.0-->m/z 292.1 for cephapirin (internal standard). The proposed method was validated by assessing specificity, linearity, limit of quantification (LOQ), intra- and inter-day precision and accuracy, recovery, and stability. Linear calibration curves were obtained in the concentration range of 0.1-20ng/mL (the correlation coefficients were above 0.99). The LOQ of the method was 0.1ng/mL. Results obtained from the validation study of goserelin showed good accuracy and precision at concentrations of 0.1, 1, 5, 10, and 20ng/mL. The validated method was successfully applied to a pharmacokinetic study of goserelin after a single subcutaneous injection of 3.6mg of goserelin in healthy white rabbits.
建立并验证了一种用于测定兔血浆中戈舍瑞林的快速灵敏的液相色谱-电喷雾串联质谱法（LC-ESI-MS/MS）。考察了影响血浆样品制备、LC 分离和 MS/MS 检测的各种参数，并对其进行了优化。酸化的血浆样品应用于 Oasis((R))HLB 固相萃取（SPE）小柱。提取的样品在氮气流下蒸发，然后用 100μL 流动相 A 重新溶解。采用 Capcell-Pak C18（2.0mmx150mm，5μm，AQ 型）柱进行分离，以溶剂 A（去离子水中的 0.05%乙酸/乙腈=85/15；v/v）和溶剂 B（乙腈）为流动相，梯度洗脱，流速为 250μL/min。LC-MS/MS 系统配备了正离子模式的电喷雾离子源。前体产物离子跃迁的多重反应监测（MRM）包括 m/z 635.7→m/z 607.5（戈舍瑞林）和 m/z 424.0→m/z 292.1（头孢匹林，内标）。通过评估特异性、线性、定量下限（LOQ）、日内和日间精密度和准确度、回收率和稳定性，对该方法进行了验证。在 0.1-20ng/mL 的浓度范围内，得到了线性校准曲线（相关系数均大于 0.99）。该方法的 LOQ 为 0.1ng/mL。在 0.1、1、5、10 和 20ng/mL 浓度下，戈舍瑞林验证研究的结果表明，该方法具有良好的准确性和精密度。该方法成功应用于 3.6mg 戈舍瑞林单次皮下注射健康白兔后的药代动力学研究。
