Srivastava A, Tripathi A K, Pandey R, Verma R K, Gupta M M
Analytical Chemistry Division, Central Institute of Medicinal and Aromatic Plants, P.O.-CIMAP, Lucknow 226015, India.
J Chromatogr Sci. 2006 Oct;44(9):557-60. doi: 10.1093/chromsci/44.9.557.
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 0.01 M (pH 3.5) phosphate buffer (NaH(2)PO(4)) 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 microg/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 microg/mL for ajmaline, ajmalicine, and reserpine, respectively, and the limits of quantitation are 19, 12, and 23 microg/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.
建立了一种采用光电二极管阵列检测的灵敏且可重现的反相高效液相色谱（HPLC）方法，用于同时定量测定蛇根木中重要的根部生物碱，即利血平、阿吗灵和阿吗碱。使用Chromolith Performance RP - 18e柱（100×4.6 mm内径）和由含0.5%冰醋酸的0.01 M（pH 3.5）磷酸盐缓冲液（NaH₂PO₄）与乙腈组成的二元梯度流动相。分析以1.0 mL/min的流速进行，检测器在254 nm波长下运行。所有生物碱的校准曲线在1 - 20 μg/mL的浓度范围内呈线性（r = 1.000）。分析的其他各个方面（即峰纯度、相似度、回收率和重复性）也得到了验证。对于这三种成分，回收率分别为98.27%、97.03%和98.38%。阿吗灵、阿吗碱和利血平的检测限分别为6、4和8 μg/mL，定量限分别为19、12和23 μg/mL。所建立的方法简单、可重现且易于操作。它对于蛇根木的评估很有用。
