Narcissus and Daffodil

Galanthamine extraction 257
whether these differences in galanthamine content resulted from extracting plants
at different stages of development or from using plants cultivated in different
regions, or were simply due to the low purity of the isolated alkaloid.
The screening described below was restricted to bulb tissue, because bulbs are
the usual commodity traded and, unlike the aerial parts of the plants, are available
in large quantities. The genus Narcissus was of particular interest, since it is indigenous
to Europe and material can easily be obtained in quantity from large
daffodil cultivars. Narcissus have been described as sources for natural galanthamine
several times (Cherkasov, 1977; Cherkasov et al., 1984, 1986, 1988, 1989).
Looking specifically for plants which would be available in large quantities, and
therefore at a low price, the screening was restricted mainly to the 20 daffodil
cultivars most commonly grown in the Netherlands (Erhardt, 1993).
The plant material was obtained from cultivars grown around Hillegom and on
Texel Island, the Netherlands. The identity of the plants was checked during the
flowering period by studying their anatomical and microscopic characteristics.
The daffodil cultivars used were identified according to ‘The International Daffodil
Checklist’ of the Royal Horticultural Society (Kington, 1989), and, as far as possible,
from reference plants and photographs.
High-performance liquid chromatography (HPLC) (Kreh, 1995) and gas
chromatography (Kreh, 1995; Kreh et al., 1995) were found to be suitable
analytical methods for the analysis of alkaloids from Amaryllidaceae. For the
quantitative galanthamine screening described below, the following HPLC
method was used:
Stationary phase: LiChrospher® 60 RP-select B, 5 µm, 250 × 4 mm column
(Merck)
Eluent A: 2.0 g sodium dodecylsulphate, 2000 ml water, 400 ml acetonitrile and
290 ml 0.05 M phosphoric acid
Eluent B: acetonitrile
Gradient: 0–25 min 25:75 A:B, 30–45 min linear to 100:0, flow 1ml/min
For a detailed description of sample preparation, analytical equipment and quantitative
evaluation, see Kreh (1995).
The results of the HPLC screening are given in Table 9.1. From the plants
tested, several narcissus cultivars were found to be rich in galanthamine.
However, in the case of the Galanthus and Leucojum bulbs tested, only low concentrations
of galanthamine were found. Differences between these values and
those reported in the literature might result from the occurrence of different
chemical races (Schulz, 1960) or from the effects of growing on different soils
(see below and Gorinova et al., 1993). Such differences were also found in
screening Narcissus ‘Fortune’ from Texel Island and from Hillegom, which have
different soil types.
Narcissus ‘Carlton’, ‘Gigantic Star’, ‘Ice Follies’ and ‘Fortune’ were found to be
rich in alkaloids and galanthamine. All of these belong to the group of ‘largecupped’
daffodils (Division 2; Kington, 1989), which might be interesting for a
further search for narcissus rich in galanthamine.
For all the subsequent studies Narcissus ‘Carlton’ (Figure 9.1) was used, since it
fulfils nearly all the requirements for galanthamine production on a large scale.
It has been cultivated for many years in large amounts in the Netherlands and