Study On The Fingerprint Chromatogram Of Oligosaccharides In Morinda Officinalis How

Radix Morindae officinalis, is one of the valuble Chinese herbs in South China. It is a common Chinese medicine recorded in the 2005's Pharmacopocia of People's Repubic of China, and its origin from family Rubiacae, Morinda officinalis How. As a Traditional Chinese Medicine (TCM), it is used for tonifying kidney, strengthening tendon and bone, and relieving rheumatic conditions, which has achieved good climical result on impotence, seminal emission, infertility, menstrual disorders, pain and cold feeling in the lower abdomen; rheumatic arthralgia; limpness of the legs. The resource of Morinda officinalis How is distributed in Guangdong, Guangxi, Fujian, and Hainan Province naturally. Guangdong Province is the primary origin of genuine Radix Morinda officinalis.Of this natural medicine, the chemical compositions including anthraquinones, iridoid glucosides, sterols, ursane-type triterpene, oligosaccharide, polysaccharides and so on. The oligosaccharide is prescribed for anti-stress, anti-depressants, immunological enhancement purpses in Chinese traditional preparations and high content. In this paper, using the oligosaccharide as target components, based on the precedent research the quality control methods have been studied now. The results were summarized as follow:Section one Literature studyWe reviewed the research of herbal textual, Species distribution data, cultivation techniques, chemical composition, pharmacological functions, quality control of the Radix Morindae officinalis by consulting many studies about Radix Morindae officinalis.Section two Experimental study Chapter one Investigate the germplasm resources of M. officinalis HowInvestigate the cultivatation, the ecology and the germplasm resources of M. officinalis in Guangdong, Guangxi, Fujian and Hainan Provine. The results indicated that the cultivating area of M. officinalis in Guangdong Provine is the maximum, small in Fujian Provine, the cultivantion almost not found in Guangxi and Hainan Provine. The cultivation of M. officinalis have demands for temperature, humidness and illumination. In this investigation, we collected the wild M. officinalis in Guangdong, Guangxi and Fujian Provine, and found the shape of the wild were different from the cultivated. Although the M. officinalis was planted in different provine, the main germplasm resource were from Guangdong Provine.Chapter two Determin the general practice physical and chemic characters of M. officinalis HowAccording to the 2005's Pharmacopocia of People's Repubic of China, the water content (Spl. IX H in Chinese Pharmacopoeia), water-soluble extract (Spl. X A in Chinese Pharmacopoeia), total ash (Spl. IX K in Chinese Pharmacopoeia) and the acid-insoluble ash (Spl. IX K in Chinese Pharmacopoeia) were checked out, the collected samples are basibasically consistent with the rule of Chinese Pharmacopoeia, but very different from each other.Chapter three Research on the oligosaccharides of M. officinalis How by HPTLC fingerprint chromatogramUsing fructose, sucrose, kestose, nystose and 1F-fructofuranosylnyslose as index components, investigating different thin layer plate, different developing solvent, different chromogenic agent, different humidity, different force-saturation times and so on, the best condition of TLC was established:20 cm×10 cm GF254 high performance thin layer plate (Merck, USA). According to the 2005's Pharmacopocia of People's Repubic of China (Spl. VI B),0.2μL of the sample solution and oligosaccharides control solution were dried for 12 h in a dryer after sucked up respectively, the plates were again developed for 80mm with Ethyl acetate-methanol-water-formic acid(12:3:2:3) as developing solvent in humidity of 47%, then were taken out, sprayed by a-naphthol-COV, warmed up 1-2 min in 105℃, finally the plate was observed immediately in daylight and the HPTLC image was documented. Chapter four Research on the oligosaccharides of M. officinalis How by HPLC fingerprint chromatogramThe chemical compositions of oligosaccharides in M. officinalis How were investigated by HPAEC-IPAD. Using kestose, nystose and 1F-fructofuranosylnyslose as index components, the optimum extract condition was established after systematically investigating many extracting methods.According to《Guideline of Traditional Chinese Medicine injections Fingerprint Technique》(provisional),Guideline of Experimental Traditional Chinese Medicine injections chromatographic fingerprint Technique》(try out) of the State Food and Drug Administration of China and related literatures the opitimal chromatography condition was gained for the HPLC fingerprint: a Hemilton RCX-10 chromatographic column at 30℃, the mobile phse consisted of 100mM NaOH (A) and 100mM NaOH,500mM NaAc (B) using a gradient program of A:0-5min,90%; 5-15min,90%-80%; 15-30min,80%-50%; 30-35min,50%; 35-40min,50%-100%. The flow rate was 0.8mL/min with a sample injection volume of 25uL.20 chromatograph peaks were chosen as the common characteristic peaks of HPLC of oligosaccharide in M. officinalis How, and defined Peak 1 as fructose, 2 glucose,3 sucrose,4 kestose,6 nystose,7 1F-fructofuranosylnyslose.The Fingerprints of oligosaccharide in M. officinal is How from Guangdong, Guangxi, Fujian, and Hainan Province, were compared with each other respectively,so the other origin in Guangdong. The fingerprint Comparisons of oligosaccharide in Commercial M. officinalis How were compared with the one of counterfeit, and the M. officinalis How with different periods of time from GAP base were compared with the wild. The fingerprint Comparisons of oligosaccharide were also carried out between different processing M. officinalis How, different parts of M. officinalis How, and different size Morinda officinalis How. The study provided helpful references for the construction of quality assessment system of M. officinalis How.