Research On Quality Control Method

Fangji powder, a Chinese Herbal Compound recorded in Chinese Veterinary Pharmacopoeia (2005), consist of 11 traditional Chinese medicines which are Radix Stephaniae Tetrandrae, Cortex Magnoliae, Poria, Cartex Cinnamomi, Semen Trigonellae, Radix Astragali, Fructus Psoralea, Rhizoma Alismatis, Agaric, Fructus Toosendan and Radix Morindae Officinalis. Duo to the various activities, Fangji powder was widely utilized in Animal Husbandry in China. According incomplete statistics, Fangji powder was manufactured by hundreds of corporation in China. So far, the QC method of Fangji powder is carried out by identification of the herbal powder of Cortex Magnoliae, Cortex Cinnamomi, Radix Astragali, Poria, Semen Trigonellae, Fructus Psoraleae and Rhizoma alismatis by microscope. There is no TLC and Quantitative standard for Fangji powder. In the present paper, quality control of fangji powder was investigated according to the requirement of improving quality standard released by committee of Chinese Veterinary Pharmacopoeia.(1)This research has established the method of TLC discrimination of tetrandrin and fangchinoline in crude Radix Stephaniae Tetrandrae material. Solid silical gel G plate (qingdao haiyang,100×200mm,0.20～0.25mm; three color separable); sample preparation:ethanol 30ml, reflux for 1 hour, set cold, filtrate, evaporated to dryness, the residues were then reconstituted with 5ml ethanol to get sample preparation; developed with chloroform-acetone-methanol(6:1:1); spray with bismuth potassium iodide’s reagent, check under visible light. On the TLC plates, sample plot was at the same position as control’s and showed the same colour. The method is simple and rapid for quality control of Fangji powder.(2)This research has established the method of TLC discrimination of Magnolol and Honokiol in crude Cortex Magnoliae material. Solid silical gel G plate (Qingdaohaiyang); sample preparation:methanol 25ml, occlude and shake for 30 min, filtrated, evaporated till the concentration is 5ml to get sample preparation; developed with benzene-methanol (27:1); spray with 1% H2SO4-vanillin reagent, heat to 100℃, check under visible light. On the TLC plates, sample plot was at the same position as control’s and showed the same colour. The method is simple and rapid for quality control of Fangji powder.(3)This research has established the method of TLC discrimination of Cinnamal in crude Cortex Cinnamomi material. Solid silical gel G plate (Qingdaohaiyang); sample preparation: ethanol 20ml, cold leach for 20 min, shake and filtrated, take the filter liquor as sample preparation; developed with petroleum ether(60～90℃)-methanol (17:3); spray with ethanol solution of dinitrophenylhydrazine, check under visible light. On the TLC plates, sample plot was at the same position as control’s and showed the same colour. The method is simple and rapid for quality control of Fangji powder.(4)A HPLC method was estimated to determination of the content of tetrandrin and fangchinoline in Radix Stephaniae Tetrandrae. The separation was performed on a waters C18 analytical column (150mmx4.6mm,5μm) with the mobile phase consisting of acetonitrile and 0.2% phosphoric acid (63:37),which was adjusted pH8.0 by triethylamine with gradient elution mode at the flow rate of 1.0 mL·min-1. The detection was set at 280nm. This method was applied to determine the two compounds in Fangji powder successfully. The standard curve were linear over the ranges of 52.55～525.5μg·mL-1 (r=0.9995) and 27.30～273.0μg·mL-1(r=0.9996), respectively. The average recoveries of tetrandrin and fangchinoline were 98.7% (RSD=1.5%) and 97.8% (RSD=2.3%), respectively. The method is simpler and faster than the quality control in Chinese Veterinary Pharmacopoeia (2005) and exhibit good accuracy, specialization and reproducibility.(5)A HPLC method was estimated to determination of the content of tetrandrin and fangchinoline in Fangji powder. The condition for separation was the same as described above. The average recoveries of tetrandrin and fangchinoline were 98.9% (RSD=1.3%) and 97.9% (RSD= 2.2%), respectively. The method is simple and rapid for quality control of Fangji powder.(6)A HPLC method was estimated to determination of the content of Magnolol and Honokiol in Cortex Magnoliae. The separation was performed on a waters C18 analytical column (150mm×4.6mm,5μm) with the mobile phase of methanol-water (7:22), The detection was set at 294nm, flow rate of 1.0 mL·min-1,30℃. This method was applied to determine the two compounds successfully. The standard curve were linear over the ranges of 20.26-202.6μg·mL-1 (r=0.9995) and 12.42～124.2μg·mL-1 (r=0.9992); the average recoveries were 96.7%(RSD=2.3%) and 96.7%(RSD=2.5%), respectively. The method is simple and rapid for quality control of Fangji powder as well.This research has established the method of TLC discrimination of Radix Stephaniae Tetrandrae, Cortex magnoliae and Cortex cinnamomi in Fangji powder, the assay method of HPLC of Radix Stephaniae Tetrandrae and Cortex magnoliae, and systematically researched the method of quality evaluation of Radix Stephaniae Tetrandrae, and supply the quality control of Radix Stephaniae Tetrandrae the science based.