Development trend of drug bioassay Author: Lv Huicheng Keywords: bioassay; Pharmacology; Drug bioassay is one of the classic drug detection majors, and the wide application of modern instrumental analysis has brought great challenges and opportunities to it. Facing the current basic situation, this paper expounds the application and development trend of biometrics specialty in the development of traditional Chinese medicine, new drug, drug safety evaluation and microbial limit inspection. [Keywords:] bioassay; Pharmacology; Drugs are special commodities, and the quality of drugs is directly related to the safety and efficacy of users. With the development of pharmaceutical industry, the methods and levels of drug detection have been continuously improved. Due to the development of modern science and technology and the mutual penetration between adjacent disciplines, the development of analytical chemistry has undergone three major changes, making analytical chemistry a modern analytical chemistry based on instrumental analysis. Facing the complex separation and analysis tasks in life science, chromatographic analysis methods came into being. The study of structure analysis, valence state analysis and crystal analysis promoted the development of spectral analysis. With the advent of the information age marked by computer application, instrumental analysis has developed rapidly, providing a variety of very sensitive, accurate and fast analysis methods for drug detection [1]. Bioassay has been greatly challenged, and its development prospect has aroused our concern about drug bioassay. 1 Characteristics and business scope of pharmacobiology 1. 1 The definition and characteristics of drug bioassay (referred to as bioassay) is a method to determine the content or safety of drugs by using the reaction between drugs (or harmful impurities in drugs) and organisms (or isolated organs and tissues). The advantage of bioassay is that the results basically meet the medical requirements and can directly reflect the curative effect or toxic and side effects of drugs, which cannot be achieved by other physical or chemical methods. Therefore, this method is still widely used in pharmacopoeias of various countries. The disadvantage of bioassay is that the inspection period is long, microorganisms have a growth and reproduction process and animals have a physiological metabolism process. The observation and analysis time is generally 2~7 days, and some experiments will be longer. There are many influencing factors, including biological differences, system operation errors and environmental conditions. Articles and utensils, animal quality, instruments and equipment will all have an impact on the results [2]. Therefore, the variety of bioassay in China Pharmacopoeia is decreasing. 1.2 business scope of drug bioassay China Pharmacopoeia is the legal drug standard, which divides drug quality control items into four categories: characteristics, identification, inspection and content. The business of bioassay mainly involves the inspection and content of Chinese and western medicines. Among them, the most items of drug safety inspection include: sterility, pyrogen, bacterial endotoxin, abnormal toxicity, safety test, acute systemic toxicity, allergic substances, irritating, hemolytic, antihypertensive substances, microbial limits and so on. Content (or titer) determination includes: microbial assay of antibiotics, biological assay of insulin, protamine sulfate, oxytocin, follicle stimulating hormone, luteinizing hormone and vasopressin. 2 Current situation of drug bioassay Due to the wide application of modern testing instruments, the variety, scope, methods and requirements of drug bioassay have also changed greatly. 2. 1 Determination of variety and range change of antibiotic content. At first, most antibiotics were determined by microbiological methods. With the development of pharmaceutical industry, the purification methods are constantly improved, and the effective components are more clear. The detection methods of many varieties have been changed to instrumental determination and chemical determination. For example, the 2000 edition of China Pharmacopoeia included about 2 19 kinds of antibiotics, among which 15 kinds of raw materials and their preparations were changed from the 1995 edition of chemical and microbial methods to high performance liquid chromatography, bringing the number of methods to 97, and only 24 kinds of microbial methods, including 9 new ones. It is predicted that at the beginning of this century, HPLC will develop into the instrument analysis method with the highest frequency in China Pharmacopoeia. It is stipulated to cancel the expired inspection of antibiotics, and the workload of microbial titer determination of antibiotics is obviously reduced. Heat source inspection of drug injection. 1942, the rabbit method was first included in the pharmacopoeia of the United States, and it was stipulated in the pharmacopoeia of all countries in the world. China Pharmacopoeia started from 1953. Since 1973, limulus lysate has been proved to be a sensitive reagent for detecting the existence of bacterial endotoxin (pyrogen). Compared with rabbits, limulus test is faster and more economical, and requires fewer samples and less workload in the operation process. Many samples can be tested every day. The 20th edition of USP (1980) contains "bacterial endotoxin test" for the first time, and USP2 1 edition (1985) contains 5 kinds of water for injection and 40 kinds of radioactive drugs. The fifth supplement of USP22 in 199 1 10 announced that rabbits were deleted from 185 and the method of bacterial endotoxin test was changed. 1995, the heat source of USP23 version 23 injection was almost replaced by bacterial endotoxin test [4]. China began to study and prepare limulus lysate in 1970s, and the Ministry of Health promulgated the bacterial endotoxin test method in 1988. This method was included in the second supplement of China Pharmacopoeia 1993, but it did not involve any varieties. On 1995, it was officially listed in the second part of China Pharmacopoeia, which stipulated more than 20 radioactive drugs such as water for injection and sodium chloride injection, and deleted the heat source test. The 2000 edition of China Pharmacopoeia was further expanded to 68 kinds. It is expected that the 2005 edition of China Pharmacopoeia will continue to add varieties, and the heat source will be replaced by endotoxin. Animal test changed to biochemical test. 2.2 The bioassay of experimental animals is inseparable from experimental animals. In the experiment, in order to reduce biological differences, improve the sensitivity of animal reactions and achieve the most satisfactory results with the least number of animals. The state attaches great importance to experimental animals. 1988, the State Council promulgated the "Management Conditions for Experimental Animals", which clearly stipulated the management, management and use of experimental animals, implemented a standard certification system and strictly managed them. According to the degree of microbial control, experimental animals are divided into four grades: ordinary animals, clean animals, animals without special pathogens and sterile animals [5]. General animal experiments must meet the standards of clean animals, with clear species, no disorder and no prescribed diseases. The higher the animal grade, the stricter the feeding management conditions and the greater the investment in facilities. Experimental animals are living reagents for experimental research. They must have purity and quantity, clear background, clear source and meet the requirements before they can be used. With the improvement of drug purity, animal experiments should be replaced by accurate chemical and physical methods or cytological methods as far as possible to test the quality of drugs and biological products, so as to reduce the use of animals. 2.3 Improvement and change of drug bioassay methods In order to shorten the operation time and reduce the experimental error, some instruments and equipment have been developed and put into use in recent years, such as: antibiotic inhibition zone tester, microcomputer pyrogen thermometer, bacterial collector, bacterial count tester, etc. , reducing the working intensity, improving the working efficiency and making the test results more accurate and reliable. The development trend of drug bioassay As a classic method, bioassay has been used up to now, which shows that it has irreplaceable characteristics and plays an important role in drug detection. Many old products use other methods to control quality, and new products will continue to be inseparable from biometrics. We should give full play to its advantages, strive to overcome its shortcomings and open up new business scope. 3. 1 The workload of microbial limit inspection is heavy. In order to control the bacterial limit of drugs, the microbial limit test was carried out for the first time in the American Pharmacopoeia 19 edition, which was included in the British Pharmacopoeia 1980 edition. China's Ministry of Health promulgated the 1990 version of the drug hygiene standards and inspection methods, and the China Pharmacopoeia 1995 version was officially included. China Pharmacopoeia (2000 edition) stipulated microbial limit standards according to dosage forms, and the scope of implementation included almost all Chinese and western medicine preparations and raw materials except injections and Chinese herbal pieces. This test has become the most widely used test item in the Pharmacopoeia, accounting for more than 80% of the laboratory workload of prefecture-level drug control institutes at present. In this inspection, there are a lot of business technologies that we need to further study, improve the detection conditions, make the data accurate, and explore new methods for rapid detection. The State Administration of Pharmaceutical Products has issued a trial standard for the inspection of pharmaceutical packaging materials, and its business scope will be further expanded, which is a new field for us to do further work and explore and study. 3.2 The role of drug bioassay in the development of traditional Chinese medicine China is a kingdom of traditional Chinese medicine. In 2000, China Pharmacopoeia included 920 kinds, including 398 kinds of Chinese patent medicines. There are 157 species with content determination, accounting for only 17% of the total. Traditional Chinese medicine has many components, impurities and interfering substances. It is difficult to detect the medicinal materials contained in compound preparations, especially large compound preparations, and there is a lot of research work to be done. Impurities in Chinese patent medicines, such as heavy metals and residual pesticides, will produce toxic and side effects to a certain extent, which will affect the safety of drugs [7]. In order to make traditional Chinese medicine preparations enter the international market, we still have a lot of work to do in the control items, contents and methods of inspection. Bioassay can be studied and discussed in toxicology and pharmacology, gradually improve the quality control standards, and play a greater role in improving the quality of preparations. 3.3 New drug research and development and safety evaluation New drug research and development is a systematic project of multidisciplinary cooperation. After obtaining a bioactive compound, R&D organizers should first organize experts in pharmacology, toxicology, pathology, veterinary medicine, genetics, biochemistry, pharmacokinetics and other fields to carry out cooperative research in the biomedical field, and manage it according to GLP. Relevant personnel from the departments of Histopharmacology, Toxicology (including general toxicology and special toxicology), Pathology, Pharmacokinetics and Toxicokinetics, Drug Analysis, Clinical Chemistry, Experimental Animals, Biostatistics, Quality Assurance, etc. discussed and evaluated in stages [8]. Bioassay can participate in research and development or give technical guidance in this regard. To sum up, drug bioassay is an important part of drug analysis and an indispensable inspection specialty. The extensive use of modern instruments will not only affect its development, but also add wings to tigers and show new prospects for drug bioassay. 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