Nair S.N.,Ramnarain Ruia College Maharashtra |
Menon S.,Institute for Advanced Training and Research in Interdisciplinary science Maharashtra |
Shailajan S.,Herbal Research Laboratory
Rapid Communications in Mass Spectrometry | Year: 2012
RATIONALE Asiatic acid (AA), a pentacyclic triterpene from Centella asiatica (L.) Urban, has shown numerous therapeutic activities. However, none of the published works to date has used high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) for determination of AA from biological fluids. Therefore, the present paper describes a sensitive HPLC/electrospray ionization (ESI)-MS/MS method for quantification of AA in rat plasma. METHODS Ammonium adduct formation of AA was essential in the development of a sensitive method with the rat plasma samples being pre-treated by a simple solid-phase extraction method. The separation was achieved on a Cosmosil C 18 column using a gradient mobile phase flow. Detection was performed using an Applied Biosystems API Q-Trap 2000 mass spectrometer equipped with an ESI source operated in positive mode with colchicine used as internal standard. RESULTS An eight-point calibration curve over the concentration range of 1.02-407.88 ng/mL for AA from rat plasma provided an optimum linear detector response (with r2 >0.9983). The mean percentage recovery (n = 3) for the low, middle and high quality control samples was 91.23 ± 1.88%, 90.36 ± 0.55% and 89.71 ± 0.21%, respectively. The intra-day and inter-day precision and accuracy of the quality control samples were within ≤5% and ±7% correspondingly. CONCLUSIONS The developed method was validated as per US FDA guidelines and applicability demonstrated by successful measurement of AA from plasma following oral administration of C. asiatica extracts to Wistar rats. The results suggest that the method could be applied to therapeutic monitoring of AA and pharmacokinetic studies in human volunteers. Copyright © 2012 John Wiley & Sons, Ltd.