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Toad Preparation of standard and quality analysis


Color oily 15 - 28 day end of white powder 29-56 (main with R) of white crystalline flakes (about 41g) 57-103 (B + c) a mixture of small column chloroform: acetone (10:1 ) acetone recrystallization of white powder. L R single crystalline monomer C (32g) 21 (2lg) 13 Extraction flow chart in Figure 1 Toad R: ResibofogeninB: BufalinC: Cinobufagin66 - 85 white, crystalline powder recrystallized in acetone + water {J monomer B (8g ) 5.32 1 crystal structure identification: white flaky crystal, melting point 12O. 5 ~ 121.0C (acetone), Liberman - Burchard reaction was positive, resulting in: one red and one blue and one green purple color reaction, and finally fading. Molecular Formula C. H. . O, molecular weight 384.22. The literature [3] reported the same. TCL started in three of the crystal system (cyclohexane: chloroform: acetone 4:3:3, chloroform: acetone 7:3, petroleum ether: ethyl acetate 6:4) Rf value in the same standard, and mixed melting point does not fall with it, the same infrared spectrum to identify the crystal Resibofogenin, the structure shown in Figure 2. Crystal 2: white granular crystal (acetone), mp 213.5 ~ 215.0C (acetone), Liberman-Burchard reaction was positive, the same infrared spectrum identical with the standard Cinobufagin, while three out system in the TLC (cyclohexane: chloroform: acetone 4:3:3, chloroform: acetone 7:3,[link widoczny dla zalogowanych], petroleum ether: ethyl acetate 6:4) the Rf values are the same, with the standard Cinobufagin not decrease the melting point of the mixture, identification of the crystal Cinobufagin . Structure shown in Figure 3. Crystal 3: white granular crystals, mp 238.5 ~ 239.0C, Liberman-BurchaM reaction was positive, the same infrared spectrum identical with the standard Bufalin. The three items to start the body in the TCI system (cyclohexane: chloroform: acetone 4:3:3. Chloroform: acetone 7:3. Petroleum ether: ethyl acetate 6: * 81.4) in the Rf values and standards the same product,[link widoczny dla zalogowanych], with the standard Bufalin not decrease the melting point of the mixture, identification of the crystal Bufalin. Structure shown in Figure 4. O Figure 2ResibofogeninHo Figure 4BufalinO3 detection limit of TLC and HPLC determination of 3.1 Determination of the sample detection limit of TLC determination of 3.1.1Resibofogenin Sample Properties: white flaky crystal. Differential response: Liberman - Burchard reaction was positive. Determination of detection limit: Resibofogenin samples with liquid: fine, said samples of 5.78 mg, placed in lml volumetric flask, add the sample dissolved in acetone, diluted to the scale. Point injection volume: 28.9g56.8gg86.7/zg115.6g. Chromatography plate: silica gel H a 1% CMC - Na board. Developing solvent: cyclohexane: chloroform: acetone 4:3:3, chloroform: acetone 7:3, petroleum ether: ethyl acetate 6:4. Reagent: concentrated sulfuric acid. Results: The three TLC showed a point to start the system and does not occur in the trailing phenomenon can explain this Resibofogenin check sample purity by thin layer chromatography to meet the requirements. 3.1.2Cinobufagin and Bufalin testing and results are as above. 3.2 Determination by HPLC sample liquid content of the sample preparation: Take appropriate amount at the point-like sample tube,[link widoczny dla zalogowanych], add appropriate amount of acetone. HPLC conditions: C18 column, mobile phase: acetonitrile: water (53:47), flow rate lml / min, sPD a 2AM wavelength 280nm. Paper speed 3cm/min. Sample: micro syringe with appropriate amount of sample fluid in the sample under the conditions of full scale measurement. RESULTS: The normalization method, in control and known standard to determine the content under the premise, the results were 99 or more. The average content of impurities: Resibofogenin almost no impurities, Cinobufagin not more than 0.70%,[link widoczny dla zalogowanych], Bufalin not more than 0.11. Description of the samples above can meet the quality requirements. Reference,[link widoczny dla zalogowanych], 1-13 Jiangsu New Medical literature compiled. Dictionary of Chinese Medicine [M]. Shanghai Science and Technology Press. [2] Xu Guojun editor. Pharmacognosy [M3. People's Medical Publishing House, [33 Jin Xiangqun. Chinese herbal medicine [J]. 1992. (3): l25-129. * 82 *

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