Hopkins N K, Wagner C M, Brisson J, Addison T E
Upjohn Company, Kalamazoo, MI 49001.
J Chromatogr. 1992 May 20;577(1):87-93. doi: 10.1016/0378-4347(92)80601-l.
A reversed-phase high-performance liquid chromatographic procedure was developed that accurately quantitates methylprednisolone (MP) and methylprednisolone acetate (MPA) in human plasma over the range 2.00-50.0 ng/ml. The internal standard, fluorometholone, was added to an aliquot of sodium fluoride-potassium oxalate-derived plasma. Samples were prewashed with hexane and extracted twice with methylene chloride. The extracts were dried with anhydrous sodium sulfate, centrifuged, and the organic layer separated and dried under nitrogen. The samples were reconstituted in mobile phase and washed an additional time with hexane before 100 microliters were injected onto a Beckman/Altex Ultrasphere ODS column with ultraviolet absorbance detection at 254 nm. Composition of the mobile phase was acetonitrile-water-glacial acetic acid (33:62:5, v/v/v). Calibration curves were obtained by unweighted, linear regression of peak-height ratios of MP (or MPA)/internal standard versus theoretical concentrations of MP or MPA using a Hewlett-Packard 3357 Laboratory Automation System. Extraction efficiencies for MP and MPA over the linear range were 86.4 and 84.7%, respectively. This method was successfully implemented for the analysis of specimens generated from a single-dose bioavailability and safety study for a new formulation of Depo-Medrol sterile aqueous suspension.
建立了一种反相高效液相色谱法，可准确测定人血浆中2.00 - 50.0 ng/ml范围内的甲基泼尼松龙（MP）和醋酸甲基泼尼松龙（MPA）。将内标氟米龙加入到由氟化钠 - 草酸钾制备的血浆等分试样中。样品先用己烷预洗，再用二氯甲烷萃取两次。萃取液用无水硫酸钠干燥，离心，分离有机层并在氮气下干燥。样品用流动相复溶，在注入100微升到配备有254 nm紫外吸收检测器的贝克曼/阿尔泰克斯超球ODS柱之前，再用己烷洗涤一次。流动相组成为乙腈 - 水 - 冰醋酸（33:62:5，v/v/v）。使用惠普3357实验室自动化系统，通过对MP（或MPA）/内标峰高比与MP或MPA理论浓度进行无加权线性回归获得校准曲线。在整个线性范围内，MP和MPA的萃取效率分别为86.4%和84.7%。该方法已成功应用于对新剂型德宝松无菌水混悬液单剂量生物利用度和安全性研究产生的标本进行分析。
