Osteitis Granules Experimental Study,

Guyanning granules , according to doctor Lee' s empirical formula in Wuhan Union Hospital ,composes of flos lonicerae japonicae , radix paeoniae rubra ,Caulis sargentodoxae, radix trichosanthis, spina gleditsiae and radix sanguisorbae and other four traditional Chinese medicine compound preparations . It had the effect of detumescence and alleviates pain. It was used for osteomyelitis, arthrophlogosis, abscess and so on in our hospital and was highly appraised by patient in several decades because of the conspicuous curative effect. The study of the preparation progress, quality standard and elementary pharmacology provided scientific foundation for becoming a new drug.The optimum extraction method was selected by the orthogonal test design. Four factors, such as decocting frequency(A),amount of water(B),soaked time(C) and decocting time(D),which had influence on extraction were chose to make a L₉(3⁴) table of the orthogonal test design. Score =(caffeotannic acid content/maximum*0.5+ extraction rate/ maximum*0.5)*100%. The sequence was arranged by influence:A＞D＞C＞B;The same factor in different level was ordered: A₂＞A₃＞A₁, B₁＞B₃＞B₂, C_＞C₃＞C₁, D₃＞D₂＞D₁ We found that the best water extraction was decocted twice with 8 times amount water, 60 minites each time by using chlorogenic acid yeild and extraction rate as index. Experiment was done as the technology mentioned above and it showed that dry slurry yield was 13.60% and chlorogenic acid extraction rate was 80.60%. The magnified experiment showed that the dry slurry yield and chlorogenic acid content were almost the same. Three factors, such as extract concentration(A), intake airflow temperature(B) and outtake airflow temperature(C),which had influence on drying were chose to make a L₉(3⁴) table of the orthogonal test design. The sequence was A＞C＞B arranged by influence on chlorogenic acid content and A＞B＞C by influence on moisture. Considering moisture content of spray powder was in extent of checkout, A₃B₃C₁ was the best extraction method. And the optimum spray drying technology was relative density 1.3, intake airflow temperature110℃, outtake airflow temperature 40℃. Experiment was done as the spray drying technology mentioned above and it showed that moisture content was 13.60% and caffeotannic acid extraction rate was 79.40%.The enlarged experiment showed that the dry extract yield and chlorogenic acid yield in the dry extract was basically the same with the result of the orthogonal test and the chlorogenic acid transfer rate was also acceptable. So the extraction technology worked.According to the qualitative analysis of Caulis sargentodoxae, radix trichosanthis, spina gleditsiae and radix sanguisorbae contained in Guyanning granules by using thin layer chromatography, it showed that there was the same colour on the corresponding place in the colour spectrum of reference medicinal substance, but there was no spot in the colour spectrum of the negative control sample. The qualitative identification had a strong specificity. The assaying method of caffeotannic acid and was established by HPLC. The chromatographic condition was this: octadecylsilane chemically bonded silica as filler, acetonitrile-0.4% phosphoric acid as mobile phase, detecting wave length was 327 nm and number of theoretical plates was not less than 5000 by chlorogenic acid. As a result, chlorogenic acid in Guyanning granules was separated well from other components. The linear extent of concentration of chlorogenic acid was 0.0025～0.04ug, r=0.9999, application of sample recovery was 99.60%(n=6),RSD=1.07%(n=6). The assaying method of peoniflorin was established by HPLC, too. The chromatographic condition was this: octadecylsilane chemically bonded silica as filler, acetonitrile-water (13:87) as mobile phase, detecting wave length was 230 nm and number of theoretical plates was not less than 5000 by peoniflorin. As a result, peoniflorin in Guyanning granules was separated well from other components, The linear extent of concentration of peoniflorin was 0.06～0.96ug, r=0.99996, application of sample recovery was 98.50%(n=6),RSD=1.61%(n=6). The repeatability of quantitative analysis was well and the method was veracious and reliable . Accelerating test and normal room temperature test came to a conclusion that all index did not change obviously and showed the tablet can maintain stability perfectly with the packaging. Initial pharmacology study showed Guyanning granules improved the ability that akaryocytes distinguished, adhered and killed bacterium and improved the recovery of granulation tissue. Guyanning granules had bacteriostatic action in both Gram-positive bacteria and Gram-negative bacteria in the experiment, especially Staphylococcus aureus ,Staphylococcus epidermidis, aeruginosus Bacillus and so on. MIC value was 0.004～0.008g/ml.