Improved HPLC Metod for the Determination of Amaryllidaceae Alkaloids

Abstract

An improved HPLC method for quantitative determination of galanthamine, lycorine and norgalanthamine was developed. HPLC separation was achieved using a reversed phase C 18 column Symmetry ® and a gradient system with acetonitrile as an organic phase and 1% (w/v) ammonium acetate buffer adjusted to pH 6.6 with acetic acid (flow rate 0.3 -0.5 ml/min.). The calibration curves were linear from 5 to 150 μg/ml (r 2 >0.99). The reliability of the proposed system was proved through reproducibility test with alkaloids extracts from in vitro shoot-clumps culture obtained from Leucojum aestivum L. and Pancratium maritimum L.

Comparative Analysis of Lycorine in Wild Plant and Callus Culture Samples ofHymenocallis littoralisby HPLC-UV Method

“…The present HPLC-UV method had a lower LOQ compared to previous methods reported on analysis of lycorine, which were in the range of 1-34 g/mL [2,7,14]. The lower sensitivity of the present method permits the quantification of minute amount of lycorine present in the ex vivo plant tissues as well as the callus culture samples.…”

“…Besides, lycorine also possesses potent antiretroviral [9], antimitotic [10,11], and cytotoxic activities [2,12]. Different analytical techniques have been described for the qualitative and quantitative determination of alkaloids in both wild plant and in vitro culture of Amaryllidaceae including GC-MS [2,11,12], spectrophotometric [2], HPTLC [11,13], and enzyme immunoassay [2]. Few HPLC methods coupled with various detection methods were described for determination of lycorine in Amaryllidaceae plants such as Galanthus species [7], Leucojum aestivum, and Pancratium 2…”

“…The Scientific World Journal maritimum [2] and Sternbergia species [14] as well as tissue culture of Pancratium maritimum [15], Narcissus confusus [10], and Leucojum aestivum [16].…”

“…It has been traditionally used in Philippines as a vulnerary [ 1 ]. Plants in the Amaryllidaceae family were reported to contain alkaloids that are known to exhibit a wide range of pharmacological activities [ 2 ]. A number of alkaloids were isolated from the Hymenocallis littoralis such as lycorine, littoraline, hippeastrine, lycorenine, tazettine, pretazettine, macronine, homolycorine, lycoramine, vittatine, and haemanthamine [ 3 ].…”

“…Lycorine, a pyrrolophenanthridine alkaloid, is one the major alkaloids found in H. littoralis [ 2 , 7 ]. It displays strong antiviral effect against poliovirus, measles, and herpes simplex type 1 viruses [ 8 ].…”