Mogatle Seloi, Kanfer Isadore
Faculty of Pharmacy, Rhodes University, Artillery Road, Grahamstown 6140, South Africa.
J Pharm Biomed Anal. 2009 Jul 12;49(5):1308-12. doi: 10.1016/j.jpba.2009.03.008. Epub 2009 Mar 20.
A pharmacokinetic interaction study between efavirenz (EFV), a non-nucleoside reverse transcriptase inhibitor used in the treatment of HIV-1 infection, and an African traditional medicine, African potato in human subjects was undertaken. This necessitated the development and validation of a quantitative method for the analysis of EFV in plasma. A simple mobile phase consisting of 0.1M formic acid, acetonitrile and methanol (43:52:5) was pumped at a low flow rate of 0.3 ml/min through a reverse phase Phenomenex Luna C(18) (2) (5 microm, 150 mm x 2.0mm i.d.) column maintained at 40 degrees C. Diclofenac sodium was used as an internal standard (IS) and EFV and IS were monitored at 247 nm and 275 nm, respectively. A simple and rapid sample preparation involved the addition of mobile phase to 100 microl of plasma to precipitate plasma proteins followed by direct injection of 10 microl of supernatant onto the column. The procedures were validated according to international standards with good reproducibility and linear response (r=0.9990). The intra- and inter-day accuracies were between 12.3 and 17.7% at the LLOQ and between -5.8 and 9.1% for the QC samples. The intra- and inter-day precision of EFV determinations were 5.1 or less and 7.2% RSD or less, respectively across the entire QC concentration range. Mean recovery based on high, medium and low quality control standards ranged between 92.7 and 94.1% with %RSD values better than 3%. Plasma samples were evaluated for short-term (ambient temperature for 6h) and long-term (-10+/-2 degrees C for 60 days) storage conditions and were found to be stable. The method described is cost-effective and has the necessary accuracy and precision for the rapid quantitative determination of EFV in human plasma.
开展了一项在人体受试者中进行的依非韦伦（EFV，一种用于治疗HIV-1感染的非核苷类逆转录酶抑制剂）与一种非洲传统药物非洲马铃薯之间的药代动力学相互作用研究。这就需要开发并验证一种用于分析血浆中EFV的定量方法。一种简单的流动相，由0.1M甲酸、乙腈和甲醇（43:52:5）组成，以0.3 ml/min的低流速泵入一根反相Phenomenex Luna C(18) (2)（5微米，150 mm×2.0mm内径）色谱柱，该色谱柱保持在40℃。双氯芬酸钠用作内标（IS），EFV和IS分别在247 nm和275 nm处进行监测。一种简单快速的样品制备方法包括向100微升血浆中加入流动相以沉淀血浆蛋白，然后直接将10微升上清液注入色谱柱。这些程序根据国际标准进行了验证，具有良好的重现性和线性响应（r = 0.9990）。在LLOQ时日内和日间准确度在12.3%至17.7%之间，质控样品的准确度在-5.8%至9.1%之间。在整个质控浓度范围内，EFV测定的日内和日间精密度分别为5.1%或更低和7.2%RSD或更低。基于高、中、低质量控制标准的平均回收率在92.7%至94.1%之间，%RSD值优于3%。对血浆样品进行了短期（室温6小时）和长期（-10±2℃60天）储存条件评估，发现其是稳定的。所描述的方法具有成本效益，并且对于快速定量测定人血浆中的EFV具有必要的准确度和精密度。
