Moreno Ronilson Agnaldo, Oliveira-Silva Diogo, Sverdloff Carlos Eduardo, Borges Bruno Carter, Rebelo Galvinas Paulo Alexandre, Astigarraga Rafael Barrientos, Borges Ney Carter
Departmento de Farmacologia, Universidade Estadual de Campinas, Campinas, SP, Brazil.
Biomed Chromatogr. 2010 Jul;24(7):774-81. doi: 10.1002/bmc.1362.
In the present study a fast, sensitive and robust validated method to quantify chlorpheniramine in human plasma using brompheniramine as internal standard (IS) is described. The analyte and the IS were extracted from plasma by LLE (diethyl ether-dichloromethane, 80:20, v/v) and analyzed by HPLC-ESI-MS/MS. Chromatographic separation was performed using a gradient of methanol from 35 to 90% with 2.5 mm NH(4)OH on a Gemini Phenomenex C(8) 5 microm column (50 x 4.6 mm i.d.) in 5.0 min/run. The method fitted to a linear calibration curve (0.05-10 ng/mL, R > 0.9991). The precision (%CV) and accuracy ranged, respectively: intra-batch from 1.5 to 6.8% and 99.1 to 106.6%, and inter-batch from 2.4 to 9.0%, and 99.9 to 103.1%. The validated bioanalytical procedure was used to assess the comparative bioavailability in healthy volunteers of two dexchlorpheniramine 2.0 mg tablet formulations (test dexchlorpheniramine, Eurofarma, and reference Celestamine, Schering-Plough). The study was conducted using an open, randomized, two-period crossover design with a 2 week washout interval. Since the 90% confidence interval for C(max) and AUC ratios were all within the 80-125% interval proposed by ANVISA and FDA, it was concluded that test and reference formulations are bioequivalent concerning the rate and the extent of absorption.
在本研究中，描述了一种快速、灵敏且经过验证的稳健方法，该方法以溴苯那敏作为内标（IS），用于定量测定人血浆中的氯苯那敏。分析物和内标通过液液萃取（LLE，乙醚 - 二氯甲烷，80:20，v/v）从血浆中提取，并通过HPLC - ESI - MS/MS进行分析。在Gemini Phenomenex C(8) 5μm柱（内径50×4.6mm）上，使用含2.5mM NH(4)OH的甲醇梯度（从35%至90%），在5.0分钟/运行时间内进行色谱分离。该方法符合线性校准曲线（0.05 - 10 ng/mL，R > 0.9991）。精密度（%CV）和准确度范围分别为：批内1.5%至6.8%和99.1%至106.6%，批间2.4%至9.0%和99.9%至103.1%。经过验证的生物分析程序用于评估两种2.0mg右氯苯那敏片剂制剂（试验用右氯苯那敏，Eurofarma公司，和参比制剂Celestamine，先灵葆雅公司）在健康志愿者中的相对生物利用度。该研究采用开放、随机、两周期交叉设计，洗脱期为2周。由于C(max)和AUC比值的90%置信区间均在巴西国家卫生监督局（ANVISA）和美国食品药品监督管理局（FDA）提出的80 - 125%区间内，因此得出结论，试验制剂和参比制剂在吸收速率和程度方面具有生物等效性。
