A rapid bioanalytical method for simultaneous determination of coroslic acid and asiatic acid in human plasma by soild phase extraction and direct injection into liquid chromatrography mass spectrometry

Neeshad P. Joshi*, Vikas V. Vaidya, Siddeshwar V. Patankar, Maharudra B. Kenkare


Asiatic acid (AA) and Corosolic acid (CA) are two naturally occurring pentacyclic triterpenes that has shown numerous therapeutic activities. However, there are not any methods for simultaneous quantitative determination of AA and CA reported so far from human plasma. The present research work involves a first of its kind rapid and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method developed and validated for simultaneous analysis of AA and CA 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 – 1000 ng/mL and 5 – 10000 ng/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. 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.


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

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DOI: http://dx.doi.org/10.21746/ijbio.2013.12.0017


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