Jadhav A S, Pathare D B, Shingare M S
Department of Chemistry, Dr. B. A. Marathawada University, Aurangabad 431004 (MS), India.
J Pharm Biomed Anal. 2007 Mar 12;43(4):1568-72. doi: 10.1016/j.jpba.2006.11.018. Epub 2006 Dec 28.
A chiral high performance liquid chromatographic method was developed and validated for the enantiomeric resolution of Valacyclovir, L-valine 2-[(2-amino-1,6-dihydro-6-oxo-9h-purin-9-yl) methoxy] ethyl ester, an antiviral agent in bulk drug substance. The enantiomers of Valacyclovir were resolved on a Chiralpak AD (250 mm x 4.6 mm, 10 microm) column using a mobile phase system containing n-hexane: ethanol: diethylamine (30:70:0.1, v/v/v). The resolution between the enantiomers was found not less than four. The presence of diethylamine in the mobile phase has played an important role in enhancing chromatographic efficiency and resolution between the enantiomers. The developed method was extensively validated and proved to be robust. The limit of detection and limit of quantification of (D)-enantiomer were found to be 300 and 900 ng/ml, respectively, for 20 microL injection volume. The calibration curve showed excellent linearity over the concentration range of 900 ng/ml (LOQ) to 6000 ng/ml for (D)-enantiomer. The percentage recovery of (D)-enantiomer was ranged from 97.50 to 102.18 in bulk drug samples of Valacyclovir. Valacyclovir sample solution and mobile phase were found to be stable for at least 48 h. The proposed method was found to be suitable and accurate for the quantitative determination of (D)-enantiomer in bulk drugs substance. It can be also used to test the stability samples of Valacyclovir.
建立了一种手性高效液相色谱法，并对伐昔洛韦（L-缬氨酸 2-[(2-氨基-1,6-二氢-6-氧代-9H-嘌呤-9-基)甲氧基]乙酯）原料药中的抗病毒药物进行了对映体拆分方法验证。在Chiralpak AD（250 mm×4.6 mm，10 µm）色谱柱上，使用含有正己烷:乙醇:二乙胺（30:70:0.1，v/v/v）的流动相系统拆分伐昔洛韦的对映体。对映体之间的分离度不小于4。流动相中二乙胺的存在对提高色谱效率和对映体之间的分离度起到了重要作用。所建立的方法经过广泛验证，证明是可靠的。对于20 μL进样体积，(D)-对映体的检测限和定量限分别为300和900 ng/ml。校准曲线在900 ng/ml（定量限）至6000 ng/ml的浓度范围内对(D)-对映体显示出良好的线性。在伐昔洛韦原料药样品中，(D)-对映体的回收率在97.50%至102.18%之间。发现伐昔洛韦样品溶液和流动相至少在48小时内稳定。所提出的方法适用于准确测定原料药中(D)-对映体的含量，也可用于测试伐昔洛韦的稳定性样品。
