Cannell G R, Mortimer R H, Maguire D J, Addison R S
Conjoint Internal Medicine Laboratory, Royal Brisbane Hospital, Queensland, Australia.
J Chromatogr. 1991 Feb 15;563(2):341-7. doi: 10.1016/0378-4347(91)80040-j.
A reversed-phase liquid chromatographic assay was developed to quantitate prednisolone, prednisone and the 20 alpha-dihydro and 20 beta-dihydro reduced metabolites of both parent compounds in tissue culture media from in vitro perfusions of the human placental lobule. Steroids were extracted from perfusate, using reversed-phase cartridges, with average recoveries of 95.2% or greater. The internal standard for the analyses was 6 alpha-methylprednisolone. In this assay cortisol coelutes with prednisolone, however, no other significant interferences were found. Assay of each steroid was linear in the range 0-1 microgram/ml. Intra-assay coefficients of variation were measured at 10 and 750 ng/ml with ranges of 3.4% (20 alpha-dihydroprednisone) to 8.8% (20 beta-dihydroprednisolone) and 4.1% (20 beta-dihydroprednisone) to 8.8% (prednisone). The corresponding inter-assay coefficients of variation were 3.3% (20 alpha-dihydroprednisone) to 9.1% (20 beta-dihydroprednisolone) and 1.9% (prednisolone) to 3.5% (prednisone). The analyses utilized two C18 columns which were linked together and maintained at 40 degrees C.
建立了一种反相液相色谱分析法，用于定量测定人胎盘小叶体外灌注组织培养基中泼尼松龙、泼尼松以及这两种母体化合物的20α-二氢和20β-二氢还原代谢产物。使用反相柱从灌注液中提取类固醇，平均回收率达95.2%或更高。分析的内标物为6α-甲基泼尼松龙。在此分析中，皮质醇与泼尼松龙共洗脱，但未发现其他显著干扰。每种类固醇的分析在0 - 1微克/毫升范围内呈线性。批内变异系数在10和750纳克/毫升时进行测定，范围为3.4%（20α-二氢泼尼松）至8.8%（20β-二氢泼尼松龙）以及4.1%（20β-二氢泼尼松）至8.8%（泼尼松）。相应的批间变异系数为3.3%（20α-二氢泼尼松）至9.1%（20β-二氢泼尼松龙）以及1.9%（泼尼松龙）至3.5%（泼尼松）。分析使用两根连接在一起并保持在40℃的C18柱。
