| PART ONE studies on the quality control about Dangguiliuhuang(DGLH) Decoction by RP-HPLCObjectives: To develop a simultaneous analysis method for nine major compound in Dangguiliuhuang Decocotion: Calycosin-7-glucoside, Acteoside, Ferulic acid, Coptisine, Baicalin, Berberine, Wogonoside, Baicalein, Wogonin by High Peformance liquid Chromatography.Methods: The chromatographic separation was performed on a Agilent TC-C18(4.6mm × 250mm)column with the gradient elution of acetointrile-0.1% phosphoric acid, the volume flow was 1.0m L/min, the detection wavelength was set at 280nmResults: 1、The nine compounds was well separated in 45 mins by gradient elution under the selected chromatographic condition after continuous optimization. 2、The calibration curves of Calycosin-7-glucoside, Acteoside, Ferulic acid, Coptisine, Baicalin, Berberine, Wogonoside, Baicalein, Wogonin are Y=29643X+11812(R2=0.9997), Y=17554X-2267.2(R2=0.9992),Y=87600X+61912(R2=0.9999), Y=17886X+4444.8(R2=0.9979),Y=54788X+31094(R2=0.9999), Y=60041X+34891(R2=0.9999),Y=62616X+31840(R2=0.9999), Y=55065X+23755(R2=0.9998),Y=225546X+176861(R2=0.9999). All calibration curves showed good linear regression(R2>0.997). 3、The relative standard deviation of precision of nine compounds range from 0.20% to 4.29%, which shows the instruments and choromatographic conditions are favorable. 4、The RSD of nine compounds in reproducibility test fluctuates between 0.12% to 1.89%, which represents good reproducibility. 5、Analyse Dangguiliuhuang Decoction sample at 0、2、4、6、8、12、24 h, and the results of RSD values showed the sample has good stability. 6、The recovery of 9 compounuds wave between 91%-106%, the results showed the sample has good recovery. Conlusion: The method determined the content of Calycosin-7-glucoside, Acteoside, Ferulic acid, Coptisine, Baicalin, Berberine, Wogonoside, Baicalein, Wogonin in Dangguiliuhuang Decoction by RP-HPLC. The method is simple, accurate, repeatable, stable, which provide a scientific grounding for quality control evaluation of Danggui Liuhuang Decoction.PART TWO Pharmacodynamics study of XEDJ granuleObjectives: In this study, we tried to investigate the antidiarrheal activity and possible mechanisms of XEDJ, and to provide references for security of drug usage in clinical practice, also to provide a new, safe and effective medicine for the infants with diarrheaMethods: The pharmacological effects of XEDJ were investigated by gastrointestinal tests, writhing test, antidiarrheal test and capillary permeability increase test. Castor oil induced acute diarrhea, rhubarb induced chronic diarrhea in mice was performed to investigate the anti-diarrhea effect, gastrointestinal propulsive experiment was performed to evaluate the gastrointestinal function, writhing test induced by glacial acetic acid was performed to study the anti-nociceptive activity, permeability increasion of celiac capillary induced by glacial acetic acid was performed to observe the anti-inflammatory effect,Results: 1, For the normal mice, after gavage doses of 3g/kg, 6g/kg, 12g/kg XEDJ granule, the average intestinal propulsion rate was 37.72±4.59 %, 32.65±5.54 and 51.76±8.73 respectively. Compared with control group, the low and middle dose group showed no significant changes on intestinal propulsion rate, while the high dose group dramatically increased the rate. 2, Oral administration of different dosage(3g/kg, 6g/kg, 12g/kg) XEDJ granule significantly inhibited the intestinal propulsion rate in mice with sthenic intestinal movement induced by neostigmine. 3, Compared with blank control group, the low dose group and middle dose group signicicantly increase serum D-xylose concentration,the results showed in a certain range of dosage, XEDJ promote absorption rate in intestinal. Whereas the high dosage reduced the D-xylose concenteation to 1.344±0.276 mg/L. 4, XEDJ granule of three different dosage(3g/kg, 6g/kg, 12g/kg) significantly decreased the total number of wet faeces in five hours in the castor oil induced acute diahrrhea mice, meantime, the high dose group(12g/kg) significantly prolonged the onset times of diarrhea and reduced the wet faeces. 5, Our data regarding the effects of XEDJ on rhubarb induced chronic diarrhea demonstrates that low dose group, middle dose group and high dose group could improve loose stool rate and diarrhea index significantly, moreover, the high dose group could inhibit the intestinal propulsion rate. 6, XEDJ granule significantly reduced the times of body tweaking compared with model group, which means the drug distinctly antagonize the pain caused by acetic. 7, Compared with blank control group, The middle dose(6g/kg) and high dose(12g/kg) group of XEDJ granule demonstrated anti-inflammatory activity by remarkblely inhibit the permeability increasion of celiac capillary.Conlusion: All the test results indicated that the drug has favorable therapeutic effects in the treatment of diarrhea, which means the traditional Chinese medicine XEDJ worth being studied further. |
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