Narcissus and Daffodil

Analysis of Amaryllidaceae alkaloids 289
and an internal standard was achieved using a 15 m capillary column within a
15 minute run time. Good linearity of response was observed for all the reference
compounds in a concentration range of 8–500 µg/ml, and concentrations
around 10 µg/ml could be quantified reproducibly using this method. Quantitative
analysis was carried out using 500 mg of dried plant material, and this
method was used to quantify alkaloids in different plant parts (Moraes-
Cerdeira et al., 1997a), and to evaluate the galanthamine content of several
Narcissus cultivars grown under experimental conditions (Moraes-Cerdeira
et al., 1997b).
Capillary gas chromatography-mass spectrometry (CGC-MS)
Kreh et al. (1995a) studied the application of CGC-MS in the detection and identification
of underivatised Amaryllidaceae alkaloids. Fifteen reference alkaloids
were subjected to CGC-MS in electron impact or chemical ionisation mode, and
their stability under column conditions was studied. All but two compounds were
found to be stable, with haemanthamine and lycorenine undergoing partial
decomposition during analysis. Derivatisation with trimethylsilylating agents failed
to improve either the sensitivity or the resolution of the analysis. Using this
method, they were able to detect and propose the structures of several new
compounds in Narcissus ‘Carlton’ (Kreh et al., 1995b).
High performance liquid chromatography (HPLC) for the
determination of Amaryllidaceae alkaloids in natural sources
High performance liquid chromatography (HPLC)
Westwood et al. (1981) investigated the feasibility of coupling HPLC with a circular
dichroism (CD) spectrometer to analyse optically active compounds with suitable
chromophores selectively. By using stopped-flow techniques, full CD spectra of
optically active compounds were recorded. An alkaloid fraction of Crinum glaucum was
chromatographed using a reversed-phase C-8 column and a mixture of methanolwater
with a trace of ammonia as the solvent. One of the major compounds in the
extract, ambelline, was detected through a comparison of the on-line recorded CD
spectrum with that obtained under standard conditions.
Evidente et al. (1983) developed a rapid quantitative analytical procedure for
lycorine using a C-18 reversed-phase column and acetonitrile:0.01 M ammonium
carbonate (47:53) as the solvent. Lycorine was analysed as the sulphate salt, and
was detected using an ultra-violet (UV) detector at 290 nm. The detection limit for
lycorine was 5 ng. These authors applied the method to quantify lycorine in crude
acid extracts of bulbs and leaves of Sternbergia lutea. Davey et al. (1998) adapted this
system under semi-preparative and analytical conditions to purify and quantify
lycorine from acid extracts of Crinum asiaticum.
Bruno et al. (1985) described an HPLC method to determine lycorine and
tazettine in the bulbs of Narcissus tazetta. The alkaloid fraction was separated on a
LiChrosorb-CN ® column using methanol:water (25:75) containing 1% Pick
reagent as the mobile phase. Compounds were detected by fluorescence, using an