Studies On Identification With Fingerprinting And Content Determination Of Nerviliae Fordii (Hance) Schltr

Nerviliae Fordii (Hance) Schltr. comes from the rhizome and whole grass of Nerviliae Fordii (Hance) Schltr in orchidaceae plants. It was recorded most early in ancient times "Ben Cao Gang Mu Shi Yi" (A Supplement to the Compendium of Materia Medica).It is cool in nature and sweet in flavor, commonly used to clear away heat and toxic materials, moisten lung, dissipating blood stasis, so it has marked effect in treating Tuberculosis, hemoptysis due to phlegm-fire, scrofula, pyogenic infection, traumatic injury, Children's Malnutri-tion, malnutritional stagnatione, psychosis, oral ulcer, acute pharyngitis and laryngitis and so on, especially in treating Children pneumonia. Nerviliae Fordii (Hance) Schltr. widely distributed in Guangdong, Guangxi, Hainan, also can be found in Sichuan,Yunnan and Guizhou.It's commonly used in Guangdon-g, Guangxi, Hongkong, Macau and Southeast Asia. Nerviliae Fordii (Hance) Sch-ltr.'s export volume is great, and the price is very high. Because of it's limited distribut-ions andresources, low reproduction coefficient in addition to unauthorized mining, resources of wild Nerviliae Fordii (Hance) Schltr. gradually exhausted.Because of Nerviliae Fordii (Hance) Schltr.'s high price, there are more and more fake and adulterants appear in the market. Civilian substitute Nervilia plicata (Andr.) Schltr. for Nerviliae Fordii (Hance) Schltr., such as Plantago depreessaWilld., Centella asiatica (L.) Urb., Ipomoea batatas Lam. and so on. commodity of Nerviliae Fordii (Hance) Schltr. can be divided into three kinds:large-leaved, middle-leaved and small-leaved, which are used disorderedly. IR and HPLC-ECD fingerprint are used to distinguish Nerviliae Fordii (Hance) Schltr., Nervilia plicata (Andr.) Schltr. and their adulterants in the study. There are great differences between Nerviliae Fordii (Hance) Schltr. Nervilia plicata (Andr.) Schltr. and their adulterants:Plantago depreessa Willd., Centella asiatica (L.) Urb., Ipomoea batatas Lam. in IR spectra identification, so they can be easily separated from each others without the second derivative test. IR spectra of Nerviliae Fordii (Hance) Schltr. in different breed and resource are very similar, they can be separated by the second derivative test, which proved that IR spectra test is a rapid and effective method of identifying Nerviliae Fordii (Hance) Schltr. and can be used in quality controlling of Nerviliae Fordii (Hance) Schltr.Plantago depreessa Willd., Centella asiatica(L.)Urb., Ipomoea batatas Lam. showed great difference in HPLC-ECD finger spectrum, so they can be easily distingushed from Nerviliae Fordii (Hance) Schltr. There are also great differences between Nerviliae Fordii (Hance) Schltr. and Nervilia plicata (Andr.) Schltr. in HPLC-ECD test. Nerviliae Fordii (Hance) Schltr. in different breed and resource can be classified and analyzed according to their chromatographic peak:Guangdong small-leaved, Guangxi small-leaved and Guangxi middle-leaved can be classified as one kind; Guangxi large-leaved can be classified as one kind, Hainan large-leaved can be classified as one kind, Guangxi large-leaved and Hainan large-leaved can also be classified as one kind. From chromatographic peak, Nerviliae Fordii (Hance) Schltr. and Nervilia plicata (Andr.) Schltr. showed great differences, which proved that they possess different chemical components, so Nervilia plicata (Andr.) Schltr. are suitable for substituting Nerviliae Fordii (Hance) Schltr., so pharmacological and Pharmacodynamic comparative study should be conducted in order to make sure nervilia plicata can substitute Nerviliae Fordii (Hance) Schltr. clinically. Fresh and dry samples of Nerviliae Fordii (Hance) Schltr. also showed great difference, which showed that Nerviliae Fordii (Hance) Schltr. collected in different period possess different chemical components.Rhamnocitrin and Rhamnazin from Nerviliae Fordii (Hance) Schltr. are antitumor and anti-oxidation active components. The result determined by HPLC-ECD showed that leaves of Nerviliae Fordii (Hance) Schltr. in different breed and resource showed great differences in content of Rhamnocitrin and Rhamnazin, which indicated that the accumulations of Rhamnocitrin and Rhamnazin are influenced by geographical environment. The contents of Rhamnocitrin and Rhamnazin from different extracted parts of Nerviliae Fordii (Hance) Schltr. are quite different:leave＞leafstalk＞root＞corm,and the content in leave is far greater than other parts, contents in root and corm are very low, especially in corm. We can substitute above-ground plant parts for whole grass if we select Rhamnocitrin and Rhamnazin as the reference component of Nerviliae Fordii (Hance) Schltr., then we can protect the resources of Nerviliae Fordii (Hance) Schltr.Nerviliae Fordii (Hance) Schltr. tissue-culture rhizome proliferation medium was optimized, the result demonstrates that medium based on MS,20g·L-1 sucrose as carbon source, add 1.0g·L-1 yeast extract or 10% of the apples, hormone:6-BA1.3 mg·L-1, PP333 1. 1mg·L-1, KT 1.5mg·L-1 as the best proliferation medium, which can promote the speed of Qingtiankui tissue-culture rhizome's reproduction. After removing the hormone 1/2MS medium, rhizome can induced more small bulbs, more than lcm in diameter of which can grow into plants that can regenerate after transplanting.Determining regeneration plants'content of Rhamnocitrin and Rhamnazi-n, we found Nerviliae Fordii (Hance) Schltr. regeneration plants have the similar active ingredients as before, which laid foundations for study on whether regeneration plants'can be substituted for natural seedling. Compa-ring content of Rhamnocitrin and Rhamnazin in different stage of tissue culture,we also found that Rhamnocitrin and Rhamnazin cannot be detected in rhizome during proliferation and induced stage until they grew into buds. This indicated that active ingredients are accumulated after rhizome differenti-ation stage and formatting leaves stage through photosynthesis.