Study On The The Resolution Of The Optical Gsssypol

OBJECTIVE To separate enantiomers of gossypol. This object include 5 important sections: To establish the analytical method of gossypol; To optimize the extraction process of Gossypol to determine the ideal extraction condition; To determine the best derivate conditional of gossypol ; Lastly,(-),(+)-Gossypol ramification were hydrolyzed by glacial acetic acid and hydrochloric acid. To obtain optical rotation method.METHODS Two methods have been established to determine the content of gossypol; Orthogonal test was conducted to study the extraction progress of Gossypol With the content of the Gossypol as the index.The 3 parameters including amount of aether(A) and petroieum(B) used for extraction. Time for extraction(C); (-)- Threo(-)-1-(p-nitophenyl)-1,3-dihydroxy propylamine-2 was found to be a useful resolving agent for racemic gossypol, Taking chloroform, Methanol, acetone, THF as impregnant to derivative reaction respectively. time of reaction is 10min,15min,20min,30min; the proportion of the two substances is 1:1 to 6:1; the column chromatogram were use to separate the derivatives; Acetoacetate : Cyclohexane=4:1 and Acetoacetate:Cyclohexane=3:2 were eluting solvent respectively.(-),(+)-Gossypol ramification were separated successfully; Lastly,(-),(+)-Gossypol ramification were hydrolyzed by glacial acetic acid and hydrochloric acid. The hydrolysis production were determined by HPLC,melting point and optical rotation method.RESULTS The HPLC and UV method, which was used to determine the content of gossypol, including external reference method and internal standard method, had very good linear correlation and it had a good degree of precision,HPLC column: ODS-C18(4.6mm×150mm,5μm), HPLC column temperature: 35℃, mobile phase: methanol-2% H3PO3=90:10,the flow rate: 1.0ml/min, wavelength: 235nm;The result shows that themethod is simple, special and accurate.The method was linear within the range of 1.37-17.12μg/ml, S.tianschanica was 1.39%, and the average recovery was 100.10%; The ideal extraction condition were A3,B1,C3,That is 12 times of aether,8 times of petroieum and the time is 40min; the column chromatogram were use to separate the derivatives. The reaction time shoaled be above 30 Minutes .The molecular proportion of the two substances must be over 2:1; Lastly,(-),(+)-Gossypol ramification were hydrolyzed by glacial acetic acid and hydrochloric acid. The hydrolysis production were determined by HPLC,melting point and optical rotation method.Conclusionthe The analytical method established was accurate and reliable.