| Ganmao qingre keli is an effective Chinese traditional formula against common cold,~-which is composed of 11 medicinal herbs: Herba Schizonepetae, Herba Menthae, Radix Saposhnikaviae, Radix Bupleuri, Herba Perillae, Radix Puerariae, Radix Plaiycodon. Semen Prunus, Radix Angelica, Herba Co~ydal is, Herba Pragmites.1 BI0AcTIvITY STUDYFour fractions were obtained from successive extraction of the crude drug with chloroform, ethyl acetate and n-butanol. According to the modem pharmacological methods for traditional diaphoretic medicines, antiinflaminative and bacteriostatic effects were used as the criteria to evaluate the bioactivities of those four fractions and volatile components of Ganmao qingre keli. It was found that ethyl acetate layer and water layer were antiinflammative while water layer was bacteriostatic. Daidzein. a compound from ethyl acetate fraction showed anti-inilammative activity.2 STUDY ON QUALITY CONTROL METHODBased on the bioactivity of Ganmao qingre keli and the known components of its composed herbs, pulegone, menthol, puerarin, daidzein, prim-O-glucosylciniifugin and 4?O- ~ -D-glucosyl-5-O-methylvisanminol were chosen as indexes of quality control.P -D-glucosyl-5-O-methylvisamminol from Saposhnikavia divaricata (Turcz.) Schischk and daidzein from Pueraria lobata (Willd.) Ohwi were identified by inC. and pulegone from Schizonepeta tenu~folia Briq., and3menthol from Mentha haplocalyx were identified by GC-MS.Four HPLC methods were developed to determine puerarin, daidzein, prim-O-glucosylcimifiigin and 4?Q- ~ -D-glucosyl-5-O-methylvisanuninol in Ganmao qingre keli.Ganmao qingre keli (with sucrose, without sugar, with lactose) were extracted with water and ethanol, centrifuged and filtered. An APEX ODS column (25OnimX4.6mm, 5pm) was used in this study.To determine puerarin, the mobile phase was 1 OOmmoIIL ammonium acetate (pH5.0)- methanol (75: 25). The flow rate was 0.8 mL/min, and the detection was set at 250 nm. The calibration curve was linear in the range of 2?0 pg/mL puerarin (r=0.999 9). The relative standard derivation was 1.1% and the average recovery was 98.6% (RSD1 .8%) in Ganmao qingre keli (with sucrose). The average contents of puerarin in Ganmao qingre keli (with sucrose, withGut sugar, with lactose) were 1.52 mg/g (RSD1.1%), 3.48 mglg (RSDO. 8%), 0.288mg1g (RSThZ3 .6%) respectively.To determine daidzein, the mobile phase was I OOmmolIL ammonium acetate pH5.0-methanol-THF (60:37:3.5). The flow rate was 0.8 mL/min, and the detection was set at 250 nm. The calibration curve was linear in the range of 0.2? .8 pg/mL daidzein (r=O.999 7). The relative standard derivation was 2.0% and the average recovery was 95.5% (RSD=l .8%) in Ganmao qingre keli (with sucrose). The average contents of daidzein in Ganmao qingre keli (with sucrose, without sugar, with lactose) were 23.5 pg/g (RSD=2.0%), 78.2 pg/g (RSD=l.4%), 13.6 pglg (RSD=3.5%) respectively.To determine prim-O-glucosylcitnifiigin, a mobile phase of methanol-40 mniol/L sodium acetate pH6.8 (35:65) was used with a flow rate of 0.8 mlimin and a detection set at 254 nm. The calibration curve was linear in the range of 0.8p.g/mL ?.2 pg/mL for prim-O-glucosylcimifugin (r=0.999 9). The relative standard derivation was 1.0% and the average recovery was 101.4% (RSD=l.8%) in Ganmao qingre keli (with sucrose). The average contents of prim-O-glucosylcimifugin in Ganmao qingre keli (with sucrose, without sugar, with lactose) were 0.1 32mgIg (RSD=1 .0%), 0.31 7mg/g (RSD 1.7%), 0.549mg1g (RSD=2.2%) respectively.To determine 4?O- P -D-glucosyl-5-O-methylvisamminol, a mobile phase of L120-methanol-THF (62:38:1) was employed with a flow rate of 0.8 mL/min, and a detection set at 254 nm. The calibration curve was linear in the4range of 0.8p.gImL ?.2 ~ig/mL for 4?O- P -D-glucosyl-5-Omethylvisamrninol (r=0.999 9). The relative standard derivation was 1.8% and the average recovery was 95.0% (RSD=1 .8%) in Ganmao qingre keli (with sucrose). The average contents of 4?O-... |
PDF Full Text Request