Quality Evaluation Research Of 6-Acety-8-Hydroxypiperidinemethyl Chrysin

Chrysin is widely distributed in nature, with anti-cancer, anti-oxidation, anti-inflammatory and other pharmacological activities. However, because of poor solubility and low bioavailability, limit its clinical application. So using Chrysin as lead compound, it is of great importance to modify its structure.6-Acety-8-hydroxypiperidinemethyl Chrysin is a totally synthetic Chrysin Mannich base derivatives by using main raw material 3-hydroxy acetophenone, benzoyl acetic ester and 4-piperidinol.In this paper, Reference standards of 6-acety-8-hydroxypiperidinemethyl Chrysin (CH-j), 8-acetyl-6-hydroxypiperidinemethyl Chrysin (isomer) and 6-acetyl Chrysin was firstly prepared, and characterized their structures by spectroscopy. Using HPLC-HRMS to speculate impurities, which the content was less than 0.15%, the mainly were M480, M465, M433, M840.In this paper, an assay method of CH-j was established by HPLC-UV. Using KF-C18 acid column; mobile phase was methanol-acetonitrile-water-methanesulfonic acid (27:24:49: 0.15); detection wavelength was at 283 nm. The theoretical plate was not less than 2000. In the range of 6.25?100 ?g/ml, the linear regression equation of CH-j was y=359557x-178921 with good linearity (r2=0.9998, n=5); and the precision RSD was 1.50%(n=6), the repeatability RSD was 1.89%(n=6). The accuracy of the assay method was verified by non-aqueous titration.The assay method of CH-j by HPLC-UV was used to establish and verified for determination of impurities. Using methanol-acetonitrile-water-methanesulfonic acid (27:24: 49:0.15) as the mobile phase to detect isomer, methanol as the mobile phase to detect 6-acetyl Chrysin. Separating degree was good (R> 1.5). Within the range of 0.625?10 ?g/ml, the linear regression equation of 6-acetyl chrysin and isomer respectively was y=578437x+238554 (r2=0.9988, n=5) and y=177696x44871.2 (r2=0.9989, n=5) with good relationship linear; the standard addition recoveries were 99.9%(RSD was 4.06%, n=9) and 99.3%(RSD was 4.92%, n=9), both were less than 5.0%, in line with the provisions; the limit of quantification were<0.5 ?g/ml.The dissolution test of CH-j tablets was established and verified through a variety of dissolution medium dissolution curves method. The log P of CH-j was 1.63, pKa was 5.3, the solubility exhibits pH dependent. Various dissolution media dissolution curve experiment results show that dissolution was 100%,90% and 6.8% respectively in 45 min in pH 1.2 hydrochloric acid solution, pH 4.5 sodium acetate-acetic acid buffer solution, and pH 6.8 phosphate buffer solution. Thus, CH-j is immediate-release pH dependent dissolution. As a result, the dissolution determination medium of CH-j tablet used pH 1.2 hydrochloric acid solution 1000 ml. The dissolution limit is 85% of the labeled amount.From what has been discussed above, the good quality evaluation method is specific, precision, accuracy, repeatability, durability analysis methods are in line with the quality requirements for the quality control of the CH-j.