Study On Quality Standard And Fingerprint Of Abelmoschus Manihot Dropping Pills

Abelmoschus manihot dropping pills are quick-acting new preparation of treating cardiovascular and cerebrovascular disease, which was prepared with Flos abelmoschus manihot as raw material by solid dispersion technology, with the guidance of the traditional Chinese Medicine theory and according to modern preparation technology, increased the bioavailability of the drug. Abelmoschus manihot dropping pills can be easily prepared and easily controlled. It has many good characters such as quick result、high efficieney and exactly quantifiable. According to the "Chinese Pharmacopoeia"(2010 edition), this study has completed the quality criteria include TLC, inspection and the initial stability, a HPLC method was also established to determine hyperin, quercetin, rutin and myricetin. To comprehensively reflect the whole chemical characteristics and intrinsic quality of the Abelmoschus manihot dropping pills, we established high performance liquid chromatography fingerprint for the quality of Abelmoschus manihot Dropping Pills, they also cover the disadvantages of quality control with few effectives in the past.Objects: In order to control the quality of Abelmoschus manihot dropping pills, we established high performance liquid chromatography fingerprint and quality standards for the quality of Abelmoschus manihot Dropping Pills, according to the character of chemical constituents and dropping pills. Sunset abelmoschus flower, hyperin and quercetin were identified by TLC. Content of hyperin, quercetin, rutin and myricetin in Abelmoschus manihot dropping pills were determined by HPLC. Has investigated initial stability of Abelmoschus manihot dropping pills. To establish high performance liquid chromatography fingerprint for the quality of Abelmoschus manihot Dropping Pills and to accumulate the date used for their quality evaluation.Methods: 1 Sunset abelmoschus flower, hyperin and quercetin were identified by TLC. 2 ①The meheod of determining the contents of Hyperoside and Rutin in Abelmoschus manihot dropping pills: the HPLC method was used for detemining the contents of Hyperoside and Rutin with a column of Kromasil C18(4.6mm×200mm,5μm), Mand with a mobile phase of acetionitrile-0.2% phosphori acid(19:81 v/v), the speed at 1.0m L/min, the detection wavelength at 360 nm,the injection volume at 10μL and the column temperature was room temperature. ②The meheod of determining the contents of Myricetin and Quercetin in Abelmoschus manihot dropping pills:the HPLC method was used for detemining the contents of Myricetin and Quercetin with a column of Kromasil C18(4.6mm×200mm,5μm),Mand with a mobile phase of methanol- acetonitrile-0.4%phosphoric acid(12:28:60 v/v/v), the speed at 1.0m L/min, The detection wavelength at 360 nm, the injection volume at 10μL and the column temperature was room temperature.3 The experience was processed according to the Chinese Pharmacopoeia(2010 version of the first general) principles of the preparation; and tested relative items of National Leechdom Standard. The weight variation and dispersion limit were determined in Abelmoschus manihot dropping pills. 4 preliminary stability study : With index of for appearance, indentification, Content and dispersion limit, Abelmoschus manihot dropping pills were exposed to light, heat and humidity to explor the factors affecting the its stability. Long term stable experiment were testsd to exam its stability. 5 HPLC separation was performed on an Inertsil ODS-3(4.6mm×250mm,5μm)by gradient elution in 80 min with acetonitrile-2m L/L formic acid as mobile phase and gradient elution at flow rate 1m L/min, the eluent monitored by a DAD detector at 360 nm, the column temperature at 35℃.Results: Abelmoschus manihot dropping pills was identified by TLC. The developing solvent was acetic ether-methanol-formic acid(10:5:3). The chromogenic agent was Al Cl3 test solution. The examination under UV lamp(365nm). The linear correlation is available between 0.990μg and 9.900μg of Hyperoside(r=0.9994, n=7). The average recovery rate was 99.54%, the RSD was 1.18%; For Rutin, the linear range was from 0.294μg– 2.940μg(r=1.0000, n=7). The average recovery rate was 100.34% and RSD was 2.38%; For Myricetin, the linear range was from 0.064μg–0.640μg(r=0.9998, n=7). The average recovery rate was 100.81% and RSD was 1.70%; For Quercetin, the linear range was from 0.030μg–0.300μg(r=0.9999, n=7). The average recovery rate was 99.64% and RSD was 2.40%. The weight variation and dispersion limit conform to the Chinese Pharmacopoeia(2010 version of the first general) principles of the preparation; and tested relative items of National Leechdom Standard. Abelmoschus manihot dropping pills were not stable under high temperature, light or humidity Circumstance and accelerated test, but it was stsble under long-term stability of 18 months. Among the obtained fingerprint, the most of the detected peaks were separated effectively, 10 batches of Abelmoschus manihot Dropping Pills were analyzed. The quanlity samples was assessed by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2004 A version).The similarities of 10 batches of Abelmoschus manihot Dropping Pills were all over 0.850,and 13 characteristic peaks of HPLC fingerprint for Abelmoschus manihot Dropping Pills for established. The accuracy, repeatability, and stability of this method were satisfied.Conclusions: The determination method of Abelmoschus manihot dropping pills is feasible and specific. The HPLC fingerprint of Abelmoschus manihot Dropping Pills is established, the conventient and high specific method could be used to qualitative analysis and control the quality Abelmoschus manihot Dropping Pills.