Studies On Extraction And Quality Control Of Total Iridoid Glycosides In The Fruits Of Ligustrum Lucidum

The source of the crude drug, Frutus Ligustr Lucidi(Chinese name, Nuzhenzi ) are the fruits of Ligustrum Lucidum Ait which is well known and frequently used in clinical practice.In the theory of Chinese traditional medicine, it has the effects of supporting the healthy energy, nourishing the liver and kidneys and can be applied to replenish the liver and kidney, improve eyesight and promote the growth of black hair. It is also used as an important component in some traditional Chinese patent medicines. Pharmacological tests show that it has the activities to improve immune function and delay the ageing process.Ever since a long time ago, it is considered that oleanolnic acid and the other compound is the active substance in the fruits of Ligustrum Lucidum Ait. However, after 20 century the world research indicate that besides the low-polar constituent oleanolnic acid, the fruits of Ligustrum Lucidum Ait is also contains water-solubility components, such as high-contents iridoid glcosides, saldroside and so on . Iridoid glcosides has many biological functions like protect liver and cholagogue, anti-hepatic fibrosis, antibiosis and dephologisticate, subdue blood-fat, cut down blood-sugar, degrade viscosity, reinforce immunological regulation, etc. Salidroside is the main active component of Rhadolia rasea L., iridoid glycosides in the fruits of Ligustrum Lucidum Ait. is consists of many compounds which have the same chemical constitution- p-hydroxyphenethyl- β -D-glucoside. So, consider iridoid glcosides in the fruits of Ligustrum Lucidum Ait as traditional Chinese drug effective elments is very important for make best of Ligustrum Lucidum Ait, have significance meaning for the development of new drug and new preparation.To establish a RP-HPLC method for determining the content of saldroside in fruits of Ligustrum Lucidum Ait. The RP-HPLC separation was accomplish on Hypersil-C₁₈ column(5μm,4.6mm×150mm) with mobile phase of methanol water (20:80) by flw rate of 0.4 mL/min.The detection wavelength was 220nm. This method is proved to be simple, rapid, accurate and suitable for the qualty control of salidroside in Ligustrum Lucidum Ait.To optimize the purified process for salidroside in Ligustrum Lucidum Ait. Compounds were extracted with ethanol and purified by macroreticular resin. The content of extration can reach 7.7%. Now salidroside was only extract from Rhadolia rasea L. which is rare and expensive. Ligustrum Lucidum Ait which is abundance in our country is adopted to extract salidroside with high contents. It provides a scientific basis for the commercial production and utilization of Ligustrum Lucidum Ait.The orthogonal design (with L9 (34) orthogonal table) was used to optimize the extraction process of total glycosides from Ligustrum Lucidum Ait, three factors in extraction were optimized, i.e., extraction times, volume of extraction system and concentration of ethanol of elutant. The scan response of the spot of the salidroside and high polar components in TLC was used as a index, the methods of extraction including reflux > cold-soaked ^ water extract and alcohol deposited determined by TLC and Scan-TLC. The result shows that by cold-soaked extraction way the optimum extraction condition obtained is as follows: alcohol concentration of 70 %, cold-soaked time of 48h and extraction times of 3 times. And the content of total iridoid glycosides in Ligustrum Lucidum Ait is the highest and the purification is the easiest.For the properties of macroreticular resin, the influencing factors such as alcohol concentration of eluant, temperature, equilibrium time, and concentration of eluant, quantity of swilled alcohol, the ratio between extraction and macroporous resin, adsorption time and the flow rate on desorption were studied and the optimal process route and conditions were suggested in this paper: Ligustrum Lucidum Ait was absorbed for 30 minutes and washed with 500mL distilled water, then eluted with 200mL of 10%-, 30%^ 50% ethanol, and finally collected the eluant of 50% ethanol to evaporate to dryness with aqueous bath. Put the collection in polyamide column, eluted with lOOmL distilled water and get the dried product was the extraction of iridoid glycosides of Ligustrum Lucidum Ait.According to the methods of extraction, we established new process chart for the extraction > separation and purification of iridoid glycosides of Ligustrum Lucidum Ait. The average content of total iridoid glycosides in the extraction was 65.7% and the average extraction rate was 10.76mg/g.We established Alkaline hydrolysis -RP-HPLC for measuring the total iridoid glycosides in the extraction of Ligustrum Lucidum Ait. That is to say after alkaline hydrolysis, determine the quantity of salidroside with RP-HPLC and calculate the content of iridoid glycosides. The concrete procedure of this method is that dissolve about 20mg extraction with 2mL 5% NaoH, then heat in aqueous bath for lh, cool down, and use diluted hydrochloric acid limestone to neutralize the pH value to 7.In this paper, we summarized the characters such as identification, determination and quality control standards to the extraction of iridoid glycosides of Ligustrum Lucidum Ait.lt can be the guarantee of the stability and security of the finished product and also be a base for the further qualitative or quantitative analytical research.