Study On Quality Standard Improvement Of Compound Shuanghua Oral Liquid

From influenza pandemic of 1918 to 1919, to the influenza A in recent years, the influenza virus, to a great extent, posed a serious threat to human health and life safety. Most influenza viruses were not sensitive to antibiotics and other Western medicine, and were likely to develop drug resistance. The study found Traditional Chinese medicine (TCM) had a good anti-influenza virus activity. And it was difficult to produce drug resistance and little side effects. Furthermore TCM mostly can mobilize the body’s immune system to resist the virus attack. Some of TCM and its preparations had been applied to clinical treatment of flu. It is urgent and meaningful for developing more TCM which had exact curative effects on the flu widely used in the clinical treatment.As the commonly used anti-influenza virus drug, Fufang Shuangfang Oral Liquid is mainly for the adjuvant therapy of acute upper respiratory tract infection, especially for the good curative effect of acute tonsillitis. Currently the qualitative identification items only include the identifications by thin layer chromatography (TLC) in the quality standards of Fufang Shuanghua Oral Liquid.And the content determination items include the determination of chlorogenic acid in the preparation by thin layer chromatography scanning (TLCS).It is necessary to improve the identifications of the whole prescription by TLC. In addition, the content determination method of active ingredients in the preparation should be established by HPLC which is of high sensitivity and good reproducibility, accurate and reliable. This paper intends to promote existing quality standards in the qualitative identification as well as the quantitative determination. It is beneficial for the more comprehensive quality control of Fufang Shuanghua Oral Liquid. And it will provide evidence for establishing comprehensive and better quality standards. The main contents and results are as follows:1 By referring TLC identification items of Lonicera japonica Thunb., Forsythia suspensa (Thunb.) Vahl, Andrographis paniculata (Burm.f.) Nees and Radix Isatidis seu Baphicacanthii in the Chinese Pharmacopoeia (2010 edition), preparation methods of sample were optimized, the conditions of TLC were modified properly, and TLC were established to identify four Chinese medicinal materials in the whole prescription. The results showed the same color spots in the TLC graph of sample existed at the corresponding positions compared with the controls and didn’t exist in the negative control. It indicated the TLC was specific, the oral liquid actually added four Chinese medicinal materials.2 By referring HPLC determination method of Lonicera japonica Thunb., and Radix Isatidis seu Baphicacanthii in the Chinese Pharmacopoeia (2010 edition), preparation methods of sample were optimized, and HPLC was established to determinate the contents of chlorogenic acid and epigoitrin which are antivirus ingredients in the Fufang Shuanghua Oral Liquid. The conditions of HPLC determinate chlorogenic acid:Dikma Diamonsil C18 chromatographic column (4.6 mm × 250 mm,5μm), acetonitrile-0.1% phosphoric acid solution (8:92) was used as mobile phase, flow rate was 1.0 mL/min, detection wavelength was 327 nm, column temperature was at 35 ℃ and injection volume was 10μL. The number of theory plates was not less than 2000 determined by the peaks of chlorogenic acid. The conditions of HPLC determinate epigoitrin:Dikma Diamonsil C18 chromatographic column (4.6 mm ×250 mm, 5μm), methanol-0.02% phosphoric acid solution (11:89) was used as mobile phase, flow rate was 1.0 mL/min, detection wavelength was 245 nm, column temperature was at 35℃ and injection volume was 10μL. The number of theory plates was not less than 3000 determined by the peaks of epigoitrin.Results showed that the peaks of chlorogenic acid and epigoitrin achieved a baseline resolution. The linear relation of chlorogenic acid was excellent within the range of 19.54μg/mL～195.40μg/m, the linear equation was Y=3×107X-37258(R2=0.9999). The linear relation of epigoitrin was excellent within the range of 0.4μg/mL～4μg/mL, the linear equation was Y= 68707X+2209.8 (R2=0.9998). The mean recoveries of chlorogenic acid and epigoitrin were 98.83% and 101.1% respectively with RSD of 1.7% and 0.51%. The mean contents of chlorogenic acid and epigoitrin in the six batches of Fufang Shuanghua Oral Liquid determined by HPLC were.1.84mg/mL and 42.83mg/mL3 By referring HPLC determination method of Lonicera japonica Thunb., Forsythia suspensa (Thunb.) Vahl, Andrographis paniculata (Burm.f.) Nees and Radix Isatidis seu Baphicacanthii in the Chinese Pharmacopoeia (2010 edition), preparation methods of sample were optimized, and HPLC was established to determinate the contents of chlorogenic acid,epigoitrin, forsythoside A,forsythin, andrographolide and dehyandrographolide which are antivirus ingredients in the Fufang Shuanghua Oral Liquid. The conditions of HPLC determination: Waters Sunfire C18 chromatographic column (4.6 mm x 250 mm,5μm), the mobile phase consisted of Methanol-0.1% phosphoric acid solution with gradient elution, flow rate was 1.0 mL/min;Chlorogenic acid,epigoitrin, forsythoside A as well as andrographolide were detected under 240nm, forsythin was detected under 225nm, dehyandrographolide was detected under 254nm;Column temperature was at 40℃ and injection volume was 10μL.Results showed resolutions and theoretical plate numbers of 6 kinds of active ingredients meet the requirement. After the methodological study, the determination method had strong specificity and good linear relationship in some linear range. And it was accurate and reliable. The mean contents of epigoitrin,chlorogenic acid, forsythoside A,forsythin, andrographolide and dehyandrographolide in the three batches of Fufang Shuanghua Oral Liquid determined by HPLC were 0.037、1.85、0.44、0.24、0.18 and 0.036mg/mL.Researches above established a simple and feasible TLC method to identify four Chinese medicinal materials and an accurate and reliable HPLC method to determinate contents of chlorogenic acid and epigoitrin in the whole Fufang Shuanghua Oral Liquid. The method established was specific and practical, and promoted current quality standards of the preparation. What is more, this paper simultaneously determined contents of six acitive ingredients in the oral liquid. This provided evidence for the establishment of a more comprehensive quality standard.