Studies On The Whole Process Of Quality Control Of Shanmei Capsule

Objective:1. To establish fingerprints of Hawthorn leaf and Rosa davurica pallâ†'Hawthorn leaf extract and Rosa davurica pall extractâ†'Shanmei capsulerespectively by HPLC chromatography,confirm the source of chromatographicpeaks of Shanmei capsule and from the whole to do the half quantitative andhalf qualitative quality control.2. To develop a method for simultaneous determination of chlorogenicacid, glucosyl-vitexin, vitexin-2″-O-rhamnoside, rutin, vitexin, hyperoside andquercetin in Hawthorn leafâ†'Hawthorn leaf extractâ†'Shanmei capsuleï¼›Todevelop a method for simultaneous determination of gallic acid, rutin,hyperoside and quercetin in Rosa davurica pallâ†'Rosa davurica pallextractâ†'Shanmei capsule. Controlling the quality of Shanmei capsule fromquantitative determination of more index components.3. To develop a method for determination of flavonoids of Hawthorn leafand Rosa davurica pallâ†'Hawthorn leaf extract and Rosa davurica pallextractâ†'Shanmei capsule by UV-Vis Spectrophotometry, and with contentdetermination of more index components together to do the quantitativequality control of Shanmei capsule from part to the overall.4. Using the method of Hawthorn leaf and Rosa davurica pall are thenegative control each other to confirm the source of chromatographic peaks ofShanmei capsule.Methods:1. Using the HPLC chromatography and the following chromatographicconditions to establish fingerprints of crude drugsâ†'extractâ†'productrespectively: Agilent ZORBAX SB-C18column (4.6mm×250mm,5μm),0.5%formic acid (A)-acetonitrile(B)-methanol(C)-tetrahydrofuran(D) were mobilephases to do the gradient elution:0~15~20~30~35~60min，A(97%~93%~89%~80%~80%~56%), B(0.5%~0.5%~0.5%~0.5%~0.5%~6%),C(0.5%~0.5%~0.5%~0.5%~0.5%~6%),D(2%~6%~10%~19%~19%~32%); The wavelength: at0~15min，the detection wavelength was260nm, thereference wavelength was360nm, at15~65min, the detection wavelength was370nm, the reference wavelength was430nm; The flow rate was1.0mL·min-1,the column temperature was30℃and the sample size was10μL. Using《Traditional Chinese medicine chromatographic fingerprint similarity ofevaluation system2004A version》of State Pharmacopeia Committee of Chinato evaluate the fingerprint similarity and confirm the common peaks’ numberin each fingerprint.2. As establishing the above fingerprints, using the external standardmethod to develop a method for content determination of more indexcomponents in crude drugsâ†'extractâ†'product respectively.3. Using the UV-Vis Spectrophotometry to develop a method fordetermination of flavonoids in crude drugsâ†'extractâ†'product by externalstandard method, rutin was reference substance,510nm was wavelength andNaNO2-Al(NO3)3-NaOH was the color development reagent.4. Using methods of reference substance comparison and negative controlto confirm the source of chromatographic peaks of Shanmei capsule.Results:1. The fingerprints of Hawthorn leaf,Rosa davurica pall,Hawthorn leafextract and Rosa davurica pall extract were marked10common peaks respe-ctively, the similarity were greater than0.9; The fingerprints of Shanmeicapsule were marked16common peaks and the similarity were greater than0.9.2. The average contents(mg/g) of each index component were as follows:Hawthorn leaf(chlorogenic acid:0.3366, glucosyl-vitexin:1.2947, vitexin-2″-O-rhamnoside:2.6933, rutin:0.0686, vitexin:0.7481, hyperoside:0.6322,quercetin:0.0290); Rosa davurica pall(gallic acid:0.4093, rutin:0.0194,hyperoside:0.0633, quercetin:0.0074); Hawthorn leaf extract(chlorogenicacid:1.4598, glucosyl-vitexin:5.1291, vitexin-2″-O-rhamnoside:10.9684, rutin:0.2864, vitexin:2.9724, hyperoside:2.3243, quercetin:0.1574); Rosa davuricapalll extract (gallic acid:1.5011, rutin:0.0517, hyperoside:0.1494, quercetin: 0.0310); Shanmei capsule(gallic acid:0.6110, chlorogenic acid:0.8562, glucos-yl-vitexin:2.5778, vitexin-2″-O-rhamnoside:5.6618, rutin:0.1571, vitexin:1.5423, hyperoside:1.2545, quercetin:0.1001).3. The average contents(mg/g) of flavonoids were as follows: Hawthornleaf:87.425, Rosa davurica pall:29.899, Hawthorn leaf extract:295.038, Rosadavurica pall extract:139.810, Shanmei capsule:221.604.4. The source of each chromatographic peak of Shanmei capsule was:Number6,7,8,9,10,11,12,13and16came from Hawthorn leaf; Number1,2,3,4,5,10,13,14,15and16came from Rosa davurica pall; Hawthornleaf and Rosa davurica pall shared Numbe10,13and16; Through thereference substance comparison, number3was gallic acid, number6waschlorogenic acid, number7was glucosyl-vitexin, number9was vitexin-2″-O-rhamnoside, number10was rutin, number12was vitexin, number13washyperoside, number16was quercetin.Conclusion:1. Developing fingerprints of crude drugsâ†'extractâ†'product respectivelyin the same chromatographic conditions can monitor the change of eachcomponent in the production process and ensure the stability of the drug.2. Developing a method for simultaneous content determination of moreindex components of crude drugsâ†'extractâ†'product can control the wholeproduction process of Shanmei capsule better.3. Developing a method for determination of flavonoids of crude drugsâ†'extractâ†'product, with content determination of more index componentsand fingerprints together to do the whole quality control of Shanmei capsule.This can provide the theory basis for playing clinical curative effect better.4. Confirming the source of chromatographic peaks of Shanmei capsulecan confirm Hawthorn leaf and Rosa davurica pall further,and can ensure theeffect of Shanmei capsule better.