Development and Validation of a Sensitive LC-MS/MS Method for Determination of Valacyclovir in Human Plasma: Application to a Bioequivalence Study

作者：Konda R K^*; Chandu B R; Challa B R; Chandrasekhar K B

来源：Acta Chromatographica, 2013, 25(4): 669-686.

DOI：10.1556/AChrom.25.2013.4.6

• valacyclovir
• LC-MS/MS
• solid-phase extraction
• bioequivalence

A simple, sensitive, and highly specific method has been developed for determination of valacyclovir (VL) in human plasma. The analytical procedure involves a solid-phase extraction method using Valacyclovir-D8 (VLD8) as an internal standard. Chromatographic separation was carried out on a reversed phase Zorbax, SB C18, 4.6 x 75 mm, 3.5 mu m column. Valacyclovir and Valacyclovir-D8 were detected with proton adducts at m/z 325.2 -> 152.0 and 333.3 -> 152.0 in multiple reaction monitoring (MRM) positive mode. The method was linear over the concentration range of 0.5-700.0 ng mL(-1). The limit of detection (LOD) and limit of quantification (LOQ) for valacyclovir were 0.2 pg mL(-1) and 0.5 ng mL(-1), respectively. The method was shown to be precise with the average within-run and between-run variations of 0.7 to 3.5% and 3.1 to 4.7%, respectively. The average within-run and between-run accuracies of the method throughout its linear range were 96.7 to 97.9 and 94.7 to 97.3%, respectively. The mean recoveries of valacyclovir and Valacyclovir-D8 from human plasma by the developed method were 99.17 +/- 10.78% and 110.84 +/- 8.74%, respectively. The method was successfully applied in bioequivalence study with 20 healthy male volunteers under fasting condition.

• 出版日期2013-12