Rapid LC-MS/MS method for simultaneous determination of coroslic acid and asiatic acid from a market formulation of Lagerstroemia speciosa leaf extract spiked in human plasma

Siddheshwar Patankar*, Ashutosh Pudage, Neeshad Joshi, Vikas Vaidya

Abstract


Asiatic acid (AA) and Corosolic acid (CA) are two naturally occurring pentacyclic triterpenes that has shown numerous therapeutic activities and are active biomarkers of the commercially important plant Lagerstroemia speciosa (L). Lagerstroemia speciosa Linn., a Southeast Asian tree has shown wide range of therapeutic activities and is therefore used in numerous dietary supplements and herbal formulations. A research article has been published previously by the same authors for simultaneous quantitative determination of AA and CA from human plasma. The present research work involves an extension of this previous work wherein a sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method has been developed and validated for simultaneous analysis of AA and CA from a market formulation containing L. speciosa leaf extract spiked in human plasma. The analytes were extracted from the matrix using a simple solid-phase extraction procedure. Glycyrrhetinic acid (GA) was used as the internal standard for both analytes. A Kromasil C18 column provided chromatographic separation of analytes followed by detection with mass spectrometry. The method involves simple isocratic chromatography conditions and mass spectrometric detection in the negative ionization mode using a Waters Xevo TQS MS/MS system. The proposed method has been validated with a linear range of 1.00 – 1000ng/mL and 5 – 10000ng/mL for AA and CA respectively. The intra-day and inter-day precision and accuracy of the quality control samples were within the acceptance criteria of ±15% for both the triterpenoids. The total elution time was about 5 min which allows higher throughput. The assay values were found to be 0.88 % and 0.31% for Corosolic acid and Asiatic acid respectively in the market formulation containing L. speciosa leaf extract.  This validated method can be applied for analysis of real samples from a bioequivalence study involving administration of formulations containing asiatic acid and corosolic acid as their active therapeutic components.

Keywords


Asiatic acid; Corosolic acid; LC-MS/MS; Solid phase extraction

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DOI: http://dx.doi.org/10.21746/ijbio.2013.12.004

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