Jemal M, Huang M, Mao Y, Whigan D, Schuster A
Bioanalytical Research, Metabolism and Pharmacokinetics, Bristol-Myers Squibb Pharmaceutical Research Institute, P.O. Box 191, New Brunswick, NJ 08903-0191, USA.
Rapid Commun Mass Spectrom. 2000;14(12):1023-8. doi: 10.1002/1097-0231(20000630)14:12<1023::AID-RCM983>3.0.CO;2-K.
A sensitive, specific, accurate and reproducible liquid chromatography/electrospray tandem mass spectrometry method was developed and validated for the quantitation of fosinoprilat in 0.2 mL of human serum. The method employed acidification (with pH 4.0 sodium acetate buffer) of the serum samples to minimize the hydrolysis of the prodrug fosinopril to fosinoprilat prior to purification by automated 96-well solid-phase extraction. The required chromatographic separation of fosinoprilat and fosinopril was achieved isocratically on a Luna C8 analytical column (2 x 50 mm, 3 microm). The total run time was 2 min. The mobile phase contained methanol and water with 10 mM ammonium acetate. Detection was by positive ion electrospray tandem mass spectrometry. The standard curve, which ranged from 2.00 to 500 ng/mL, was fitted to a 1/x(2) weighted linear regression model. Fosinoprilat quality control (QC) samples used to determine the accuracy and precision of the method were prepared in human serum at concentrations of 5.00, 200, 400 and 1000 ng/mL. The assay accuracy was within 8% (dev). The intra- and inter-assay precisions were within 6 and 3% (RSD), respectively. Fosinopril QC samples used to gauge the rate of hydrolysis of fosinopril to fosinoprilat during the assay procedure were prepared in human serum at 500 ng/mL. The hydrolysis of fosinopril to fosinoprilat was </=1%. 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 with the drug.
建立了一种灵敏、特异、准确且可重复的液相色谱/电喷雾串联质谱法，用于定量测定0.2 mL人血清中的福辛普利拉。该方法在通过自动96孔固相萃取进行纯化之前，采用血清样品酸化（用pH 4.0的醋酸钠缓冲液），以尽量减少前体药物福辛普利水解为福辛普利拉。在Luna C8分析柱（2×50 mm，3μm）上通过等度洗脱实现了福辛普利拉和福辛普利所需的色谱分离。总运行时间为2分钟。流动相包含甲醇和含有10 mM醋酸铵的水。检测采用正离子电喷雾串联质谱法。标准曲线范围为2.00至500 ng/mL，拟合为1/x(2)加权线性回归模型。用于确定该方法准确性和精密度的福辛普利拉质量控制（QC）样品在人血清中以5.00、200、400和1000 ng/mL的浓度制备。测定准确度在8%（偏差）以内。批内和批间精密度分别在6%和3%（相对标准偏差）以内。用于评估测定过程中福辛普利水解为福辛普利拉速率的福辛普利QC样品在人血清中以500 ng/mL制备。福辛普利水解为福辛普利拉的程度≤1%。这种转化程度在分析给药后血清样品时产生的误差很小，因为已知此类样品中前体药物的含量与药物相比很低。
