The Study Of Lvjiaobuxue Granule Of The HPLC Chromatographic Fingerprint

Objective:To establish the HPLC fingerprint analysis for evaluation of quality of Lvjiaobuxue Granule,provide a reference for the scientific evaluation of its quality.Methods:①Firstly, the herbs in the prescription of Lvjiaobuxue Granule were analyzed for its base source, trait identification and quality standards;②Secondly, the best extraction and purification method was determined by the results that we analyzed HPLC fingerprint chromatogram results of the different methods of extraction the herbs;③HPLC fingerprint comparison study on the medicinal herbs such as angelica, astragalus, codonopsis, stractylodes, rehmannia in the prescription of Lvjiaobuxue Granule and the intermediates of Lvjiaobuxue Granule was explored the correlation of the Lvjiaobuxue Granule and intermediates, medicinal herbs;④Lvjiaobuxue Granule was established L-glutamic acid, L-hydroxyproline, glycine, L-alanine, L-arginine, L-proline as index components, and was determined its content;⑤astly, it was analyzed Lvjiaobuxue Granule on HPLC fingerprint of Total fingerprint peak calibration.Results:①he donkey-hide gelatin, astragalus, codonopsis, rehmannia, atractylodes, Angelica herbs of Lvjiaobuxue Granule are in line with the requirements of Chinese Pharmacopoeia2010;②It was established the best extraction method that the berbs extracted by water and n-butanol and chromatographic conditions were:column:Inertsil ODS2-C18column (4.6mm X250mm,5μm); mobile phase:Aphase of acetonitrile and the B phase was0.1%phosphoric acid solution; detection wavelength:215nm; column temperature:30℃; flow rate:1.0mL· min-1; injection volume:10μL, linear gradient elution;③Beside atractylodes, the other herbs in Lvjiaobuxue Granule and intermediates have the same the significant peaks of corresponding to the finished product.④In17batch of Lvjiaobuxue Granule, the average contents of L-glutamate was0.743%, L-hydroxyproline was0.880%, glycine was1.817%, L-alanine was692%, L-arginine was0.579%, L-proline was1.190and six kinds of amino acids were5.900%. Its contents were higher than the corresponding provisions of the indicators.⑤The17batch of Lvjiaobuxue Granule were in line with requirements. The research established the HPLC fingerprint detection methods of Lvjiaobuxue Granule that19common peaks and seven chemical compositions were identified. The sample similarity of the17batch of Lvjiaobuxue Granule was above0.90which was required in technical regulation.Conclusion:It was established the best extraction method that the berbs extracted by water and n-butanol, purificated by water extraction and alcohol precipitation, and method of the conditions of the HPLC fingerprint. The method precision and reproducibility was good, stable, reliable, and quality control of blood particles can be used to Lvjiaobuxue Granule.