| Hand foot and mouth disease(HFMD)is caused by entervirus infection, occurs more often in children under 5 years of age, and can cause herpes in hand, foot, mouth and other parts of the body, severe cases can lead to death. In recent years, hand foot and mouth disease frequently broke out, upward trend displayed, and it has become one of the acute infectious diseases which affect children’s physical health. Currently, there is no preventive medicine and potent western medicine against intestinal virus medicine, mainly symptomatic treatment and organ support therapy. Compared with western medicine, although the effect of traditional Chinese medicine preparation of antiviral is slow, the safety factor is high, the curative effect is accurate, and the effects are comprehensive, no obvious side effects. In view of this situation, the more highlights of TCM syndrome differentiation and treatment of flexibility, effectiveness and pertinence of advantage are needed, the inheritance and carry forward of the advantages and characteristics of the traditional Chinese medicine treatment are of the vital role for improving the efficacy of hand foot and mouth disease.Through the long-term clinical practice, Qingre kangdu oral liquid has a good clinical effect. But because the traditional decoction of taste is a bit poor, the dosage is large, not convenient for children to take, therefore, this project aims to explore the establishment of a pure Chinese medicine – Qingre kangdu oral liquid preparation for treating pediatric HFMD preparation technology optimization and quality standards, and quality control method. By Optimizing the orthogonal design extraction and concentration process of Qingre kangdu oral liquid, to improve the medicinal herbs decocting rate, avoid the disadvantages of traditional decoction, improve the bioavailability of the drug. By exploring the quality standard of Qingre kangdu oral liquid, to establish the analytical method with berberine hydrochloride, which is the main active ingredient for quality control index. Through the establishment of optimization and quality standard of extraction process, to provide new ideas for clinical treatment of hand foot and mouth, change the present state of western medicine treatment of hand foot and mouth disease, which has no specific prescription, and to provide safe and effective drugs to reduce the occurrence of severe hand foot mouth disease rate.Part One Study on Preparation Technology of Qingrekangdu Oral liquidObjective:By the comprehensive weighted score method and orthogonal design test, to determine the best extraction process optimization prescription medicines and to establish the optimal preparation process of Qingre kangdu oral liquid.Methods:By using orthogonal design method, and set the relative density and TLC Scanning Determination of berberine hydrochloride as the indexes, to probe the influence of extraction time, the amount of water(Times), effect of extraction times on extraction effect, and synthesize the scoring method to optimize the experimental data, determine the optimum extraction process.Results: Set the relative density and berberine hydrochloride as the indexes, the influences of various factors on the comprehensive score: the effects of the amount of water extract on berberine hydrochloride are maximum; the extraction times followed, the effects of extraction time on the extraction of berberine hydrochloride are minimum. Prescription medicine decocting 2 times, the first extraction with 10 times amount of water, boiling 1 hours, extracting for second times with 8 times amount of water, boiling 1 hours, decoction, filtration, the filtrate was concentrated to a relative density of 1.05-1.08 solution, centrifuging, adding proper amount of syrup and sodium benzoate, stir well, filling, sterilization, and obtained.Conclusion: The introduction of the weighted comprehensive evaluation method and the orthogonal design can optimize the reasonable preparation process, and make the production process simple, save the cost, enhance the operability, and it is feasible.Part Two Establishment of Quality Standard of Qingre kangdu oral liquidObjective: To establish the method for content determination by high performance liquid chromatography method for the determination of berberine hydrochloride in Qingre kangdu oral liquid. To establish a quality standard for Qingre kangdu oral liquid, provide analysis method with berberine hydrochloride, which is the main active ingredient, as the quality control index of Qingre kangdu oral liquid.Methods:(1) Distinguish the prescription drug cogongrass rhizome, uncaria, Coptis by TLC.(2) Optimization of the chromatographic conditions: select the appropriate stationary phase, mobile phase, and the detection method of chromatographic conditions, with berberine hydrochloride peaks and the other peaks separated completely.(3) System suitability test: under the above conditions, to determine whether the chromatographic peak component separation degree, the number of theoretical plates, symmetric factor meet the requirements.(4) Specificity test: Test whether the negative samples are interferential under the chromatographic conditions.(5) Preparation of standard curve: the minimum detection limit test, precision test, stability test and repeatability test.(6) Recovery test.(7) sample determination.(8)the establishment of quality standard of Qingre kangdu oral liquid.Results:(1) TLC identification of cogongrass rhizome, uncaria and Coptis is negative interference free, included in the text, TLC identification of dandelion has negative interference, so it is not included in the text.(2)Methods: extraction with hydrochloric acid- methanol(1:100) as solvent, ultrasonic 40 min for the best extraction method.(3) Optimization of the chromatographic conditions: chromatographic columns for the SHIMADZU C18(4.6 * 150 mm, 5μm); acetonitrile-0.05mol/L potassium dihydrogen phosphate(25:75)as mobile phase elution; the detection wavelength was 265nm; the flow rate was 1.0ml/min.(4) System suitability test: under the above conditions, the number of theoretical plate of berberine hydrochloride of chromatographic peak is more than 3000, and the adjacent chromatographic peak separation degree is greater than 1.5.(5) the specificity test: under the above chromatographic conditions, the negative control product without interference for the samples.(6) preparation of standard curve: a good linear relationship in the range of 0.05-5μg /ml sample amount of berberine hydrochloride showed, the correlation coefficient is 0.9999.(7) the minimum detection limit test of berberine hydrochloride: minimum detection limit was 0.025μg /ml.(8) the precision test: the effective peak area of Berberine Hydrochloride by RSD was 0.13%.(9) stability test: effective peak area of Berberine Hydrochloride by RSD was 0.6%, the sample in 8 hours stability.(10) the repeated test: Berberine Hydrochloride by peak area calculated repeatability test of RSD was 1.8%.(11) recovery test: berberine hydrochloride and the average recovery was 101%, RSD was 1.86%.(12) sample determination: under the established chromatographic conditions, to determine the the content of berberine hydrochloride Qingre kangdu oral liquid from different batches of determination, and calculate the content of effective components in the sample.Conclusion: Determine the content of berberine hydrochloride in Qingre kangdu oral liquid by HPLC. The method has short analysis time, strong specificity, wide linear range, good precision and accuracy and other advantages, andcan be used for the quality control of Qingre kangdu oral liquid, provides the quality control method with scientific and feasible way for ensuring the quality of preparation. |
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