Jemal M, Mulvana D E
Bioanalytical Research, Metabolism and Pharmacokinetics, Bristol-Myers Squibb Pharmaceutical Research Institute, New Brunswick, NJ 08903-0191, USA.
J Chromatogr B Biomed Sci Appl. 2000 Mar 10;739(2):255-71. doi: 10.1016/s0378-4347(99)00551-4.
A sensitive, specific, accurate and reproducible LC-MS-MS method was developed and validated for the simultaneous quantitation of the prodrug fosinopril and its active drug fosinoprilat in human serum. The method employed acidification of the serum samples to minimize the hydrolysis of fosinopril to fosinoprilat prior to purification by solid-phase extraction to isolate the two analytes and the two internal standards from human serum. The extracted samples were analyzed by turbo ionspray LC-MS-MS in the positive ion mode. Chromatography was performed on a polymer-based C18 column (Asahipak ODP PVA-C18, 2x50 mm) using gradient elution with methanol and 10 mM ammonium acetate, pH 5.5. The calibration curve, 1.17 to 300 ng/ml, was fitted to a weighted (1/x) linear regression model. Serum quality control (QC) samples used to gauge the accuracy and precision of the method were prepared at concentrations of 5.00, 100, 250 and 500 ng/ml of each analyte. The inter-assay accuracies were within 6% (DEV) for both analytes. The intra- and inter-assay precisions were within 7% and 11% (RSD), respectively, for both analytes. The hydrolysis of fosinopril to fosinoprilat during sample processing was < or = 6%. This degree of conversion would cause little error in the analysis of post-dose serum samples since such samples are known to contain low levels of the prodrug compared to the drug.
开发并验证了一种灵敏、特异、准确且可重复的液相色谱-串联质谱法，用于同时定量测定人血清中前体药物福辛普利及其活性药物福辛普利拉。该方法采用酸化血清样品，以尽量减少福辛普利在通过固相萃取纯化之前水解为福辛普利拉，从而从人血清中分离出两种分析物和两种内标。提取的样品通过涡轮离子喷雾液相色谱-串联质谱在正离子模式下进行分析。色谱分离在基于聚合物的C18柱（Asahipak ODP PVA-C18，2x50 mm）上进行，使用甲醇和10 mM醋酸铵（pH 5.5）进行梯度洗脱。校准曲线范围为1.17至300 ng/ml，采用加权（1/x）线性回归模型拟合。用于评估该方法准确性和精密度的血清质量控制（QC）样品，每种分析物的浓度分别为5.00、100、250和500 ng/ml。两种分析物的批间准确度均在6%（偏差）以内。两种分析物的批内和批间精密度分别在7%和11%（相对标准偏差）以内。样品处理过程中福辛普利水解为福辛普利拉的程度≤6%。由于已知与药物相比，给药后血清样品中前体药物的含量较低，因此这种转化程度在分析给药后血清样品时不会造成太大误差。
