| DF spray, which is made of Potentilla freyniana Bomm and several other kinds of adjuvants, is a liquid spray of traditional Chinese medicine, clinically for treating herpes zoster by lower fever, detoxication, astringing sore, haemostasis. The author has made an systematic experimental study on its produing process, quality standard and primary stability, and has made some research on crude drug's quality standard as well.1. Study on Preparation ProcessingThe content of activity tannin was determined by UV Spectrophotometric; the content of catechin was determined by HPLC. Use the total extract yield and active tannins and effective components of catechin extracted as a synthetic index to study the effect of solvent, size, alcohol concentration, extraction time, extraction temperature, solvent dosage on extraction. And on this basis, the conditions of extraction technology and Percolation extraction Process were optimized by L9 (34) orthogonal test. Extraction results showed that the preferred conditions: cashing the crude herbs into 8-10 head size, and soaking it with 60% alcohol 100mL for 24 hours, and adding 60% alcohol 10 times the amount of drug, and percolating and extracting with flow rate at 1.0mL·min-1·kg-1, and collect the Percolation liquid.In the point of producing technology, some study has been done on how to choose the concentration of dissolvent, the variety and dosage of corrigent, and that of cosolvent. The method and order of adding adjuvant had great effect on solving result. On this basis, the content of activity tannin was determined by UV Spectrophotometric, and the content of catechin was determined by HPLC. The ethanol's concentration, menthol's dosage, and propanediol's dosage as three main factors forming agents were optimized by L9 (34) orthogonal test, we choose synthetic index of appearance and content to investigate processing conditions. The the optimum process was as follow: with 40% alcohol to solve the mixture of menthol and propylene glycol, adjust the concentration of alcohol to 50% after adding the mixed adjuvant, medicine: Solution =1∶12, and all targets were ideal.2. Study on Preparation's Quality StandardsGallic acid, catechin and menthol in preparation were indentified by TLC. Result: all separated spots in a clear map, and negative control without interference. We has investigated relative density, pH, heavy metal arsenic salt, ash content, acid-insoluble ash, the concentration of alcohol, the content of uniformity, and ejection disparity with the methods in PH. The results showed: Preparation relative density in 0.94～0.95, pH in the range of 4.5～5.5, the total jetting amount of a bottle should not less than 26.00g, and 10 times of the weight per injection should be in weight of 1.31g～1.60g. On the basis of the purification of sample, the content of activity tannin was determined by UV Spectrophotometric at wavelength 760nm, and the content of catechin was determined by HPLC. Chromatographic conditions: column: Agilent Zorbax SB-C18 (4.6 mm×250mm, 5μm), the mobile phase: methanol-water-acetic acid (15∶81∶1), flow rate=0.9ml/min, temperature of 30℃, detection:278nm wavelength. The content of alcohol and propylene in preparation were determined by GC, chromatographic conditions: column: wax capillary column (30m×0.319mm×0.25μm), the sample inlet temperature: 240℃, the detector temperature: 240℃, shunt procedures warming: initial temperature of 45℃, maintaining 4.5 min, 10℃per minute rate rose to 200℃, maintaining 3min. The content of menthol in preparation were determined by GC, chromatographic conditions: column: wax capillary column (30m×0.319mm×0.25μm), Inlet temperature of 225℃, the detector temperature of 240℃, shunt procedures warming: initial temperature of 100℃, maintain lmin, 5℃per minute rate rose to 160℃, maintaining 1min. Determination: tannins linear concentration range of 0.928μg～4.640μg; Catechin linear concentration range of 0.64μg～3.20μg. Stability in 10 hours RSD=3.21%; Ethanol linear concentration range of 1.58×10-2μg～4.74×102μg; propylene glycol linear concentration range of 1.608×10-3μg～4.824×10-3μg; Menthol linear concentration range of 0.02μg～0.2μg, Stability in the 24 hours RSD=1.72%; The precision of the method above RSD were 0.70%, 2.77%, 0.60%, 2.37%, 1.77%, respectively, and reproducibility's RSD were 2.16%, 1.73%, 1.93%, 1.32%, 2.24%. Apart from determination of tannin's content, the average recovery were 99.00%, 99.88%, 101.99%, 100.72%, respectively, and RSD=1.40%, 0.94%, 1.93%, 2.67%. The above quantitative method is accurate, simple, reliable, and can be used for the determination of corresponding components in preparation.3.Study on the primary stability of preparationIn order to inspect stability of DF spray, a room-temperature test has been taken for three month. The results show that every index mark was normal in the experiment period except for the slightly decrease of the content of tannin and catechin. This phenomenon imply that should add some anti-oxidant to prevent active component's oxidation.4. study on the quality standards of Potentilla freyniana BornmGallic acid, catechin in Potentilla freyniana Bornm were indentified by TLC. The results showed that all spots were separated, and the negative control without interference. Heavy metals ingredients, arsenic salt, ash and acid insoluble ash were checked, and relevant quality standards were to be formulated. The results showed heavy metal in every gram of medicine was less than 20ppm, the content arsenic salt was less than 2ppm, and did not detect acid insoluble ash.The content of activity tannin was determined by UV Spectrophotometric at wavelength 760nm, and the content of catechin was determined by HPLC. Chromatographic conditions: column: Agilent Zorbax SB-C18 (4.6mm×250mm, 5μm), the mobile phase: methanol-water-acetic acid (15∶81∶1), the velocity=0.9mL/min, temperature of 30℃. Result: Tannins linear concentration range of 0.928μg～4.640μg; Catechin linear concentration range of 0.64μg～3.20μg, Stability in the 12 hours RSD=2.48%, the average recovery was 99.38%, RSD=1.05%; The precision of two methods above RSD were 0.32%, 1.14%, and reproducibility's RSD ere 1.97%, 1.92%. These results show quantitative method is accurate, reliable, and can be used in the content determination. Besides, six different sources of crude medical samples were be detected by HPLC, and the atlas were collected to meet the relevant technical requirements. We did related analysis between the individual sample mean and the standard data, and analyze the genetic distance between the samples. Sample cluster analysis were carried out by use of SPSS10.1 software and rendered a hierarchical clustering map. Results can be seen from map: three batches from EnShi County and one batch from DaZhou County casted off closer relationship, but the products from Xuan'En and LiChuan County as the alternative form of medicine production were noticeably different from the former. Study on the fingerprint, not only reflected the overall characteristics of the sample, It also reflects the relationship among the samples, this research provide a theoretical reference to all varieties of quality evaluation.Through the research of DF spray, Preparation processing research and evaluation of the liquid spray's quality are necessarily representative. It find a solution to the quality problems which exsited in the Chinese liquid spray medicine, and lay a foundation to improve the quality of Chinese liquid sprays medicine, and rich liquid spray medicine research as a reference. |
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