The Study On Quality Control And Stability Of The Extract Of Gouteng And Its Preparation

The research on quality control and stability of drugs is an important part of drug pre-clinical studies. During the process of study on drug quality standards, an all-around check of drug quality is needed to guarantee the safety and reliability of drugs, and guarantee the smooth progress of pre-clinical and clinical research, as well as the safety of post-marketing drugs. Drug storage conditions and validity duration of drugs are determined by the stability study.In the guidance of "Chinese Pharmacopoeia" and national relevant technical regulations on quality control of drugs, a systemic study on the quality standard and stability of Uncaria rhynchophylla extract and its preparation was conducted, the quality standard (draft) of Uncaria rhynchophylla extract and its preparation was established. The thesis is divided into the following four parts:1、The study on quality standard and stability of Uncaria rhynchophylla extractIn this part of the thesis, the quality standard of Uncaria rhynchophylla extract was investigated according to Chinese Pharmacopoeia and relevant technical specifications of the drug quality control. According to the property of Uncaria rhynchophylla extract, we selected the appropriate methods for its quality control. The items of solubility, dentification, examination of inorganic impurities, moisture, weight of moisture absorption, residue on ignition et al were studied to determine the appropriate test methods and results. The results comply with the limits.The methods of residual organic solvents determination was established. The results indicated that chloroform and ethanol residues of Uncaria rhynchophylla extract met the requirements of Chinese Pharmacopoeia. In addition, the principal active components of Uncaria rhynchophylla extract were identified, and the total alkaloids, principal active components, four representative alkaloids were determined. Total alkaloid contents of three batch samples were determined larger than50%by the acid stain colorimetry. The possible constructions of principal active components were deduced, and the total amount of identified active components was between44.8%-51.7%. The four representative alkaloids were determined between31.52-32.66%by HPLC.Besides, the stability of Uncaria rhynchophylla extract was investigated according to the guiding principles of drug stability test. Impact factors test, accelerated test and long-term test were carried out respectively. The items investigated included the sample characteristics and the content of four representative alkaloids. By the impact factors test, it was found that Uncaria rhynchophylla extract is very sensitive to light, temperature, relatively insensitive to humidity. After the accelerated test at25℃, humidity of60%, a slight agglomeration occurred, the contents of four representative alkaloids were not below the limits. After the12-months’ long-term test at6℃, there is no change in sample characteristics, and the contents of four representative alkaloids were not below the limits. According to the current situation, Uncaria rhynchophylla extract can be stored under this condition.Uncaria rhynchophylla extract should be stored in a dark, dry and refrigerated condition.2、The study on quality standard and stability of Uncaria rhynchophylla extract preparationIn this part of the thesis, the quality standard of Uncaria rhynchophylla extract preparation was investigated according to Chinese Pharmacopoeia and relevant technical specifications of the drug quality control. The tests on appearance, uniformity of dosage units, dissolution and microbial limit were conducted according to the requirements for capsules in Chinese Pharmacopoeia. The test results all met the requirements. Besides, the study on identification and assay was conducted. The contents of Uncaria rhynchophylla extract and four representative alkaloids in the preparation were respectively determined by ultraviolet spectrophotometry and HPLC. No content results are below the limits.Besides, the stability of Uncaria rhynchophylla extract was investigated according to the guiding principles of drug stability test. Impact factors test, accelerated test and long-term test were carried out respectively. The items investigated included characteristics, dissolution and content of four representative alkaloids. By the impact factors test, it was found that the preparation was sensitive to light, temperature and humidity. After the accelerated test at40℃, humidity of75%, there is no change in sample characteristics, the contents of four representative alkaloids decreased by5.7-6.9%, and the dissolution decreased approximately by8%. After the accelerated test at30℃, humidity of65%, there is no change in sample characteristics, the contents of four representative alkaloids decreased by3.2-4.6%, and the dissolution decreased approximately by5%. After the6-months’ long-term test at25℃, there is no change in sample characteristics, and the contents of four representative alkaloids and the dissolution were not below the limits.According to the current situation, Uncaria rhynchophylla extract preparation can be stored under this condition.Uncaria rhynchophylla extract should be stored in a dry, cool and dark condition.3、The quality standard drafts of Uncaria rhynchophylla extract and its preparation and the drafting instructionsThe quality standard drafts of Uncaria rhynchophylla extract and its preparation and drafting instructions were formulated and drafted according to the research of the first two chapters. This chapter provides norms and basis for quality control of Uncaria rhynchophylla extract and its preparation.4、Study on ionic liquid-microwave assisted extraction for the alkaloids in Uncaria rhynchophyllaIn this part of thesis, ionic liquid coupled with microwave-assisted extraction (IL-MAE) method was applied to extract active ingredients from Uncaria Rhynchophylla, and the effects of four process parameters (concentration of ILs, liquid-solid ratio, extraction time and pH value) on extraction efficiency were optimized by mono-factor test, using the content of representative alkaloids (rhynchophylline, isorhynchophylline, corynoxeine and isocorynoxeine) as index. The optimal extraction process parameters was determined by uniform design. The optimal extraction conditions were as follows: extraction solvent of0.54mol·L-1[C4MIM]BF4aqueous solution, pH adjusted to1.42; solid/liquid ratio of1:100g·mL-1, extraction time of8min. The total content of representative alkaloids was2.52mg·g-1. The method is proved to be simple, reasonable, feasible and of good reproducibility.