Studies On The Intestinal Absorptive Kinetics In Rats Of Tongsaimai Pellets

Tongsaimai Pellets is a new pharmaceutical preparation that was redeveloped fromTongsaimai Tablets. The latter was routinely utilized in clinic to treat ischemic cerebralapoplexy and was listed in the National Protective Catalogue of Traditional ChineseMedicine. The constituent crude drugs of Tongsaimai Pellets include radix astragali, radixangelica, flos lonicerae, radix scrophulasise, radix glycyrrhizae, et al. It has the effect ofremove heat, nourish Yin, activate blood circulation and decrease blood stasis. It can beutilized to treat ischemic cerebral apoplexy, cerebral thrombosis and its sequel, venousthrombosis, blood vessel blocking caused by arteriosclerosis, et al.This thesis studied the chemical compounds and the intestinal absorptioncharacteristics of the bioactive compounds in Tongsaimai Pellets. The main contents andproductions can be divided into three parts:PartⅠLiterature investigation of the bioactive compounds in Tongsaimai Pellets,including: "clinical prevention and cure of ischemic cerebral apoplexy & research onChinese crude drugs in Tongsaimai Pellets"; "the advancement of intestinal absorption fromthe different points of views".The above work paved the way for the succeeded research ofTongsaimai Pellets.PartⅡInvestigation of the chemical compounds and the intestinal absorptioncharacteristics of the bioactive compounds in Tongsaimai Pellets, including:1. By comparing with standard chemicals&crude drug extractive, 7 compounds inTongsaimai Pellets were identified by HPLC. The peak which tR=10.218min waschlorogenic acid, the peak which t_R=23.689min was ferulic acid, the peak whicht_R=25.010min was liquiritin, the peak which t_R=51.963min was liquiritigenin, the peakwhich t_R=55.032min was cinnamic acid, the peak which t_R=58.589min was harpogoside,the peak which t_R=71.455min was monoammonium glycyrrhetate.2. The method which can quantify the amount of chlorogenic acid, ferulic acid,liquiritin, liquiritigenin, cinnamic acid, harpogoside, monoammonium glycyrrhetate simultaneously in Tongsaimai Pellets by HPLC was built up. The result displayed thatwithin the range of 53.6～268μg·mL^-1, chlorogenic acid had a good linear relationshipwith r=0.999 9; within the range of 34.2～171μg·mL^-1, ferulic acid had a good linearrelationship with r=0.999 8; within the range of 100.4～502μg·mL^-1, liquiritin had a goodlinear relationship with r=0.999 7; within the range of 12.6～63μg·mL^-1, liquiritigenin hada good linear relationship with r=0.999 7; within the range of 12～60μg·mL^-1, cinnamicacid had a good linear relationship with r=0.999 7; within the range of 5.4～27μg·mL^-1,harpogoside had a good linear relationship with r=0.999 7; within the range of 276.2～171μg·mL^-1, monoammonium glycyrrhetate had a good linear relationship with r=0.999 8. Theaverage recovery rate of chlorogenic acid was 98.78%, RSD was 0.65%（n=5）; The averagerecovery rate of ferulic acid was 98.64%, RSD was 0.84%（n=5）; The average recovery rateof liquiritin was 100.81%, RSD was 1.80%（n=5）; The average recovery rate ofliquiritigenin was 101.13%, RSD was 2.16%（n=5）; The average recovery rate of cinnamicacid was 99.46%, RSD was 2.02%（n=5）; The average recovery rate of harpogoside was99.58%, RSD was 1.62%（n=5）; The average recovery rate of monoammoniumglycyrrhetate was 100.80%, RSD was 1.70%（n=5）.In Tongsaimai Pellets of 3 batches, the content of chlorogenic acid is 1.1821%; thecontent of ferulic acid is 0.3017%; the content of liquiritin is 2.7047%; the content ofliquiritigenin is0.2574%; the content of cinnamic acid is 0.0757%; the content ofharpogoside is 0.2009%; the content of monoammonium glycyrrhetate is1.1899%.3. The intestinal absorption characteristics of general flavones, chlorogenic acid,ferulic acid, liquiritin, liquiritigenin, cinnamic acid, monoammonium glycyrrhetate ofTongsaimai Pellets were investigated with the model of in situ intestinal absorption in rats.（1）. Stability of general flavones, chlorogenic acid, ferulic acid, liquiritin, liquiritigenin,cinnamic acid, monoammonium glycyrrhetate of Tongsaimai Pellets in blank intestinalperfusate after incubation. The results were: 99.63% of general flavones; 85.24% ofchlorogenic acid, owing to instable nature; 97.56% of ferulic acid; 98.04% of liquiritin;91.43% of liquiritigenin, a sprinkle was degradated; 101.45% of cinnamic acid; 102.29% ofmonoammonium glycyrrhetate. They all fit the detection of biological specimen.（2）. After incubation for 4h in Tongsaimai pellets solution, The concentration ofgeneral flavones, chlorogenic acid, ferulic acid, liquiritin, liquiritigenin, cinnamic acid,monoammonium glycyrrhetate of gut wall&silica gel was 90.79%、93.27%、95.21%.98.74%、94.21%、85.13%、96.46%. （3）. Within 5～20 g·L^-1, the absorption rate constants Ka of general flavones,chlorogenic acid, ferulic acid, liquiritin, liquiritigenin, monoammonium glycyrrhetate atdifferent concentrations of Tongsaimai Pellets were not changed by concentrations. Theabsorption of these 6 compounts fit linear dynamics. The relationship of the absorptionamount and dose was consistent with direce ratio. The absorption mechanism of these 6compounts was passive diffusion. The absorption amount of cinnamic acid at upperconcentration (20g·L^-1) was the lowest. Cinnamic acid was considered as being carried bycarriers. Ihe absorption was saturated.（4）. There was no significant different in the absorption of ferulic acid, liquiritin&monoammonium glycyrrhetate. General flavone was absorbed faster in duodenum;chlorogenic acid was absorbed faster in colon; cinnarnic acid was absorbed faster in theupper site of intestine.4. The absorption of chlorogenic acid &ferulic acid in single herb was the fastest.Their absorption may be promoted by other components of herb. The absorption ofliquiritin, liquiritigenin, cinnamic acid& monoammonium glycyrrhetate of monomer wasthe fastest. Because of the diversity of Tongsaimai Pellets’ components, their absorptionwas inhebited.5. The absorption mechanism of ferulic acid, the effect of P-gp catastaltic, SAA,chlorogenic acid, liquiritin, liquiritigenin, cinnamic acid&monoammonium glycyrrhetate onthe absorption of FA were investigated.（1）. The absorption rate constant of FA was not changed by concentrations. Theabsorption mechanism of FA was passive diffusion.（2）.The absorption amount of FA was increased with Hydrochloric Verapamil according toduodenum, jejunum, ileum. There were significant difference between FA and FA withHydrochloric Verapamil in the intestinal absorption of FA. Ferulic acid was the substrate ofp-glycoprotein. Hydrochloric Verapamil and FA binded P-gp sites competitively. Thus, the absorptionof FA was augmented by Hydrochloric Verapamil.（3）. There was no significant influence on the intestinal absorption of FA withpolysorbate.（4）. The effects of chlorogenic acid, liquiritin, liquiritigenin, cinnamic acid,monoammonium glycyrrhetate on the intestinal absorption of FA were evaluatedrespectively. Chlorogenic acid greatly inhibited the intestinal absorption of FA （P＜0.05）,depressing gradually according to duodenum, jejunum, ileum. Liquiritin inhibited the intestinal absorption of FA in duodenum, while facilitating in ileum, without significantinfluence on jejunum （P＜0.05）. Liquiritigenin had the similar effect, but weaker thanliquiritin. Cinnamic acid inhibited the intestinal absorption of FA （P＜0.05） approaching to1/2. Monoammonium glycyrrhetate slightly promoted the intestinal absorption of FA inileum, without influence on duodenum&jejunum.PartⅢDiscussion...