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Anoectochilus (Orehidaceae), is an important Chinese medicinal herbs. It has been proved to have analgesic (pain releasing), blood sugar reducing, diuretic, anti-inflammatory and liver protecting effects by traditional medical practitioners. Thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) were applied to the compositions analysis on samples of these two species obtained from various culture periods and plant parts. According to HPLC analysis, roots contain highest component, follow with leaves and stems. A green spot at Rf 0.11 under 254 nm was only detected in Indonesia species by TLC. A blue fluorescent spot at Rf 0.3 was found in the stem and leaf samples but not in the root fraction. Two yellow-brown spots at Rf 0.61 and 0.74 by P-anisaldehyde/H2SC4, respectively, were observed only in one month old liquid culture. Further analysis will be conducted to detect difference on the active ingredients between species and their applications.
There are two species of Anoectochilus in Taiwan, including A. formosanus , A. koshunensis. Since they are morphologically alike and can only be differentiated by the characteristics of the flower apparatus. Thus it is very difficult to tell them apart beyond the blossom period. The newly developed random amplified polymorphic DNA (RAPD) technique has been used as a powerful tool for the species identification. Twelve collections from each of A. formosanus, A. koshunensis and two putative hybrids of A. formosanus X A. koshunensis were used to screen RAPD makers between species. Among 40 random primers screened, two primers did not amplify any DNA products, 5 primers amplified polymorphic bands in both A. formosanus and A. koshunensis. Twenty six out of 40 primers can only amplify RAPD in either A. formosanus or A. koshunensis. Eight primers from 26 that produced 18 RAPD makers, 9 were only present in A. formosanus, and the other 9 were only observed in A. koshunensis.
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