Identification The Reducing Carbohydrates In Extract Solutions Of Traditional Chinese Medicine By Pre-column Derivatization HPLC

A lot of active constituents were found in traditional Chinese medicine (TCM) during the modern research, but most active constituents are not as active as them in the decoction mixture. One possible reason is the single compound lost its synergistic function in the decoction. some active constituents undiscovered in decoction, especially polar constituents, is another possible reason. The possible polar constituents in decoction of TCM are amino acids, peptides, proteins, organic acid, saccharides et al. Polysaccharides and oligosaccharides which displaying notable biological activities, become the hot subject of research. There are a lot of studies concerning polysaccharides in TCM, but oligosaccharides are less studied. The studied oligosaccharides focus on Morinda officinalis How, Panax ginseng, Rehmannia glutinosa. It is reported that the oligosaccharides in Rehmannia glutinosa can applied in treatment memory impairment in mouse. The oligosaccharide in Rehmannia glutinosa can decrease plasma glucose level and plasma lipid level of hyperglycemic rats and allxan-induced diabetic rats. The reasons that oligosaccharides are less studied are their polar property and no UV absorbance. The polar property of oligosaccharides leads them difficulties to separation. No UV absorbance leads them difficulties to detection. If unpolar functional group with significant UV absorbance labelled to the oligosaccharides, the separation and detection of them will be much improved. Pre-column derivatization is applied widely in study of polysaccharides in TCM. Studies focus mainly on monosaccharides composition of polysaccharides in decoction of TCM. The reducing carbohydrates which contain monosaccharides and oligosaccharides in decoction of TCM are less studied.In this dissertation, seven reducing carbohydrates were used as the standard, reacting with PMP to produce the PMP derivatives that were analyzed by HPLC. The conditions of derivatization reaction were optimized by studying the effects of amount of PMP and NaOH, reaction time, reaction temperature, extraction solvents, times and dosage of extraction.D-Glucose, D-Galactose, D-Mannose, Maltose and Lactose, L-(+)-Arabia and D-Xylose were good linearity over the range of 0.72-144pg·mL-1(r=0.9995), 0.72-144μg·mL-1(r=0.9997),0.72-144μg·mL-1(r=0.9997),1.44-288μg·mL-1(r=0.9992), 0.6-120μg·mL-1(r=0.9992). The detection limit is 0.18μg·mL-1、0.18μg·mL-1、0.18μg·mL-1 0.36μg·mL-1、0.15μg·mL-1。This method is simple, reliable and the average recovery was good, RSD<5%. The method is suitable used for analysis of types and contents of reducing carbohydrates in decoction of TCM. The results showed that glucose is a ubiquitous composition in decoction of TCM, and the content is higher than others. Some unknown reducing carbohydrates are also existed in the decoction of TCM. Apply the method in this dissertation cannot determine the specific structure of unknown constituents, but certain is that they belong to reducing carbohydrates.On the basis of Chromatographic behavior, Some of them are oligosaccharides possibly.