Quantitative analysis of lycorine in Sternbergia species growing in Turkey

Abstract

Plants of the family Amaryllidaceae are well known not only for their ornamental value but also for the alkaloids they produce. Some of these alkaloids exhibit interesting pharmacological and/or biological properties. However, the most extensively studied effects are those of non-specific inhibition, such as antiviral and antitumour activities [1]. Sternbergia species, a member of this family, was found to contain lycorine as a major alkaloid. Sternbergia is represented by 6 taxa in Turkey [2].In our previous studies, six alkaloids (lycorine, homolycorine, galanthamine, haemanthamine, haemanthidine, and tazettine) were isolated, chemically characterized, and analyzed by HPLC in different species of Sternbergia [3][4][5]. The analgesic, antiinflammatory, antimicrobial, and antioxidant activities of some Sternbergia species and lycorine were also investigated by us [3,[5][6][7].This paper is a part of our ongoing studies [3-7] on this genus in which we attempt to quantify of lycorine from five species of the genus Sternbergia.On reviewing the literature regarding the analysis of lycorine, we found a lack of HPLC systems for studying lycorine. Furthermore, an HPLC procedure for the separation and quantification of lycorine from the acidic extract of Sternbergia lutea has also been reported by Evidente et al. [8].Good separation and determination of Sternbergia lutea ssp. lutea in bulbs was performed by using the mobile phase consisting of ammonium carbonate and acetonitrile (85:15 v/v) and a Supelcosil LC-18 column (250×4.6 mm i.d., 5 μm Supelco, Belleforte, PA, USA) at a flow rate of 1 mL/min and column temperature 24°C. Chromatograms were plotted by a Diod-Array Detector (DAD) at the wavelength 292 nm.Limits of detection (LOD) were established at a signal-to-noise ratio (S/N) of 3. Limits of quantification (LOQ) were experimentally verified by six injections of lycorine at the LOD and LOQ concentrations. The LOD was calculated to be 2.5 μg/mL and the LOQ was calculated to be 7.5 μg/mL for lycorine (Table 1).The precision of the method (intra-day variations of replicate determinations) was checked by injecting lycorine nine times at the LOQ level. The precision of the method, expressed as the RSD % at the level, was 0.646 % for lycorine (Table 2).Quantitative determination of lycorine in the bulbs of Sternbergia species was carried out by RP-HPLC using the external standard method.The assay results of Sternbergia species are shown in Table 3. Standard solutions of lycorine were added to the plant extracts and injected at each time. The area of peaks corresponding to the standards were increased to prove the presence of these compounds. Their percent mean and standard deviation values are summarized in the same Table 3.

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.…”

“…Lycorine absorbed maximum UV wavelength at 325 nm, which was selected as detection wavelength. The wavelength selected differs from previous reports which ranged between 287 and 292 nm [ 2 , 7 , 14 ]. Lycorine was eluted in the present HPLC method within 5 min and the total run time was 10 min.…”

“…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].…”

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

Quality control and anticholinesterase activity determinations on Sternbergia sicula

“…(23,24). A detailed literature search reveals that lycorine is the most quantified alkaloid in Sternbergia species (25,26). Recently, we have quantitatively determined the content of lycorine in several Amaryllidaceae species including S. sicula by HPLC-DAD analysis (27).…”