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Purpose: Semen is made legume seeds and soybeans mature Artemisia annua, mulberry leaves from the processing of fermented products. Has a solution form, Chufan, Xuanfa Yu heat and other effects, for colds, chills and fever, headache, irritability, chest tightness, trouble sleeping imaginary embolism treatment. Current clinical use SSP is based on \lack of stability, and thus directly affect the clinical safety and efficacy of the drug. In order to ensure stable and controllable Semen quality, the study of the processing technology for SSP quantization process parameters, establish scientific and standardized processing technology. While the SSP in raw soybeans, and accessories for TLC mulberry and Artemisia annua, and the establishment of multi-SSP method for the determination of index components to develop and improve the scientific and reasonable standards of quality control and evaluation of objective SSP provide the scientific basis for the quality and methodology. Methods: The study used a processing technology univariate tests to SSP isoflavones daidzein and genistein content of indicators, respectively, mulberry and processing technology in the proportion of the amount of Artemisia annua, mix soy concoction when The relative density, cooking time, fermentation temperature, fermentation time, and then a little stuffy steaming time and time and other parameters were investigated; using L9 (34) orthogonal to mulberry leaves and total flavonoids in Artemisia annua for the control targets, Select the effect of total flavonoids of mulberry and Artemisia annua boiling time, boiling times, add water to investigate factors tested, preferably a decoction of Artemisia annua mulberry and technology, combined with the actual situation of the validation process decoction test, validation test to determine mulberry and decoction of Artemisia annua process. Eventually preferred the SSP's processing technology, clear and quantified process parameters. In order to examine the feasibility of processing technology, the results of the study conducted three pilot batches, testing the indicators meet the requirements for the standardization of SSP provide the basis for production and quality control. 2 Quality standard of processing technology to standardize processed into SSP as the research object, identification and determination from TLC aspects of research, establish and improve Semen quality standards. 2.1 TLC method using thin-layer chromatography, respectively, of the test solution preparation method, the agent system, viewing conditions were optimized methods to establish SSP in raw soybeans, and accessories mulberry and qualitative methods of identification annua . 2.2 To establish a HPLC method for the determination method and UV method for simultaneous determination of SSP in isoflavones daidzein and genistein content approach and methodological studies conducted; through the determination of the samples and to develop appropriate content limits, the final Semen quality standards formulated draft, can serve as an important basis for evaluation of the quality of SSP. Results: 1 Processing Technology concocted by the findings of the inspection process parameters, optimized processing technology is: Take Mulberry 90 g, Artemisia annua 100 g, adding about 18 times the amount of crude drug boiling water three times, each time for 1 hour, filtration, the solution was concentrated to a relative density of 1.10 ~ 1.12 g/cm3, mix soy 1000 g in the net, soon after exhaustion, steam 1.5 hours, removed, slightly dry, with fried mulberry leaves, Artemisia annua slag coverage, placed in a temperature of (30 ± 2) ℃ incubator within the fermentation stuffy 6-8 days to pass on yellow, remove, remove the dregs, washed and dried, that was. By measuring the three batches of these samples, the indicators showed that the quality of the finished product are in compliance with the requirements described processing technology developed reasonably practicable. 2 results 2.1 Quality Standards 2.1.1 Semen TLC results in the identification of raw soybeans through the test, respectively, daidzein and genistein as a control reference substance in the test products for chromatography, chromatography with reference substance corresponding The position was the main spots of the same color, we can prove the existence of SSP in raw soybeans. 2.1.2 Semen in accessories mulberry leaves, Artemisia annua identification and tested, respectively, mulberry and Artemisia annua control medicine as a control, the use of the same TLC plate with a developing solvent system identification methods for the same view, in the chromatogram of the test in the reference drug chromatography corresponding position, was the main spots of the same color can be proved in Semen accessories mulberry leaves, Artemisia annua existence. 2.2 Determination 2.2.1 Determination of daidzein and genistein content results Chromatographic conditions and system suitability test octadecylsilane bonded silica as a filler, methanol and 0.1% acetic acid water (50:50) as the mobile phase, detection wavelength was 261 nm, a flow rate of 1 mL / min. Number of theoretical plates genistein peak should not be less than 3000. Preparation of standard solutions were accurately weighed daidzein and genistein appropriate reference substance, dissolved in methanol and diluted to 1 mL, respectively, each containing daidzein and genistein 10.3μg and 9.92μg solution as the reference solution . Preparation of the test solution to take the powder about 0.5 g, accurately weighed, set stoppered Erlenmeyer flask, precision dilute ethanol 20 mL, said that given the weight, ultrasound treatment 50 min, remove and let cool, then weighed weight of dilute ethanol weight loss supplement, shake, filtration, the filtrate obtained, that is, too. Determination of precision drawing reference solution and the test solution 10μL, into the liquid chromatograph to measure, that is. This product is on dry goods, including daidzein (C15H10O4) and genistein (C15H10O5) shall not be less than 0.063%, respectively, and 0.040%. 2.2.2 UV - visible spectrophotometry isoflavones results Preparation of standard solutions were accurately weighed amount of genistein reference substance, dissolved in ethanol and diluted to contain 72.0μg per 1 mL of solution as the reference solution . Preparation of standard curve genistein precision drawing reference solution 1.0 mL, 2.0 mL, 3.0mL, 4.0 mL, 5.0 mL set 50 mL volumetric flask, dilute to the mark with ethanol, shake, according to the UV - visible spectrophotometry (Appendix VA). The absorbance was measured at 261 nm, the concentration of abscissa, absorbance of the vertical axis, the standard curve. Determination Take the powder of about 0.5 g, accurately weighed, set stoppered Erlenmeyer flask, precision dilute ethanol 20 mL, said that given the weight, ultrasound treatment 50 min, remove and let cool, then weighed and the weight of dilute ethanol weight loss supplement, shake, filtration, the filtrate 5 mL precision drawing set evaporating dish, evaporated water bath with 15 mL of water quantitatively transferred home separatory funnel, extracted with ethyl acetate 25 mL 6 times, The extracts were combined bath evaporated, dissolved in ethanol and diluted to 25 mL volumetric flask, shake, precision drawing 3 mL set 10 mL volumetric flask, dilute to the mark with ethanol, shake, absorbance was measured according to the law, computing, that is, too. This product is on dry goods, including SSP isoflavone genistein to (C15H10O5), shall not be less than 0.223%. Conclusion: a processing technology through the SSP to quantify all aspects of research, to develop specific and feasible technical parameters established process to produce quantitative indicators, to achieve standardization of Semen processing technology, the process is stable and suitable for the production of SSP processing. 2 trial was the first to establish the use of the same TLC plate with a developing solvent system the same view method excipients mulberry and herbs Semen annua TLC method, and its main raw material for the qualitative identification of soybean, the method is simple , reproducible, specific for the SSP identify the authenticity of herbs provides an accurate and effective method. 3 to establish a HPLC method and UV method for simultaneous multi-index constituents on the SSP method for the determination of the method is simple, stable, reliable and can be used for multi-index Semen Determination of the contents, and then on the SSP to conduct a comprehensive and scientific quality control. This topic standardized Semen processing technology, to develop and improve the existing level of quality standards, in order to provide the basis for SSP pharmacopoeia standards, which effectively controlled the Semen quality stability, in order to ensure the safety of clinical use and reliability to provide a scientific basis.
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