Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
44, Number 9, October 2006, pp.557-560
Quantitative
Determination of Reserpine, Ajmaline, and Ajmalicine in Rauvolfia
serpentina by Reversed-Phase High-Performance Liquid Chromatography
A. Srivastava, A.K. Tripathi, R. Pandey, R.K.
Verma, and M.M. Gupta
Analytical Chemistry Division, Central Institute of Medicinal and Aromatic Plants,
P.O.-CIMAP, Lucknow 226015, India
A sensitive and reproducible reversed-phase
high-performance liquid chromatography (HPLC) method using
photodiode array detection is established for the simultaneous
quantitation of important root alkaloids of Rauvolfia serpentina,
namely, reserpine, ajmaline, and ajmalicine. A Chromolith Performance
RP-18e column (100 x
4.6-mm i.d.) and a binary gradient mobile phase composed of 0.01M
(pH 3.5) phosphate buffer (NaH2PO4) containing 0.5% glacial acetic
acid and acetonitrile are used. Analysis is run at a flow rate
of 1.0 mL/min with the detector operated at a wavelength of 254
nm. The calibration curves are linear over a concentration range
of 1–20 µg/mL (r = 1.000) for all the alkaloids.
The various other aspects of analysis (i.e., peak purity, similarity,
recovery, and repeatability) are also validated. For the three
components, the recoveries are found to be 98.27%, 97.03%, and
98.38%, respectively. The limits of detection are 6, 4, and 8
µg/mL for ajmaline, ajmalicine, and reserpine, respectively,
and the limits of quantitation are 19, 12, and 23 µg/mL
for ajmaline, ajmalicine, and reserpine, respectively. The developed
method is simple, reproducible, and easy to operate. It is useful
for the evaluation of R. serpentina.
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