Centella asiatica (L.) Urban is a traditional herbal medicine used in Asiatic countries, and is
commonly used to treat various wounds, leprosy, tuberculosis and lupus diseases. In this
work, a new method based on microwave assisted extraction followed by liquid chromatography–
diode array detection–electrospray ionization multistage tandem mass spectrometry
analysis has been developed for the identification and quantification of biologically
active constituents in C. asiatica, including asiatic acid, asiaticoside and madecassoside. The
separation was performed on an Agilent Eclipse C18 column (150 mm 9 4.6 mm i.d., 5 lm)
with gradient elution of acetonitrile and 0.1% aqueous acetic acid within 50 min. Detection
was performed at 205 nm. The calibration curves showed good linearity (r2 > 0.9992). The
limits of detection ranged from 1.2 to 1.6 lg mL-1. The intra- and inter-day precision was
less than 3% and the recovery of the assay was in the range of 95.4–106.8%. The method
was successfully applied to the quantification of the three constituents in different samples of
C. asiatica. The results indicated that the developed method could be considered to be a
simple, rapid and reliable method for the quality evaluation of C. asiatica. The samples were
also analyzed on a liquid chromatography–electrospray ionization–time-of-flight mass
spectrometry system to confirm the identification results.