Hochhaus G, Fröehlich P, Hochhaus R, Möllmann A, Derendorf H, Möllmann H W
College of Pharmacy, University of Florida, Gainesville 32610, USA.
J Pharm Biomed Anal. 1998 Sep;17(8):1235-42. doi: 10.1016/s0731-7085(98)00022-3.
A combined HPLC/RIA procedure is described for the selective determination of budesonide (BUD) in plasma. The assay involves the extraction of plasma or serum samples with ethylacetate, consequent HPLC separation of intact budesonide from cross-reacting metabolites on a C8 reversed phase column, collection of the budesonide containing fraction and determination of budesonide immunoreactivity with the budesonide antiserum. The method was accurate, sensitive (IC50 value of 0.9 ng ml-1) and reproducible (intra- and inter-day less than 15%) with a limit of quantification of 0.133 ng ml-1 (RSD < 25%). The evaluation of a limited number of clinical samples after oral administration of budesonide by both the HPLC/RIA procedure and a direct RIA using the same antiserum differed in average by a factor of 2, with the ratio of HPLC/RIA-RIA results declining as a function of time. Thus, this ratio might be a suitable indicator for probing for the ratio of budesonide and overall metabolites on a semi-quantitative level.
描述了一种用于选择性测定血浆中布地奈德（BUD）的高效液相色谱/放射免疫分析（HPLC/RIA）联合方法。该测定方法包括用乙酸乙酯萃取血浆或血清样品，随后在C8反相柱上通过HPLC将完整的布地奈德与交叉反应代谢物分离，收集含布地奈德的馏分，并用布地奈德抗血清测定布地奈德免疫反应性。该方法准确、灵敏（IC50值为0.9 ng/ml-1）且可重复（日内和日间变异均小于15%），定量限为0.133 ng/ml-1（相对标准偏差<25%）。通过HPLC/RIA方法和使用相同抗血清的直接RIA对口服布地奈德后的有限数量临床样品进行评估，平均相差2倍，HPLC/RIA与RIA结果的比值随时间下降。因此，该比值可能是在半定量水平上探究布地奈德与总代谢物比值的合适指标。
