| Cinnamon is a kind of staple in Chinese traditional medicine. There are a lot of prescriptions consisted of Cinnamon or Cassia twig. For example, Such two prescriptions were increased in the first part of Chinese pharmacopoeia 2000. So, It is necessary to study the pharmacokinetics of the effective compositions in Cinnamon.Objects: To extract cinnamic aldehyde from cinnamon and determine its content; To study the biotransformation of cinnamic aldehyde and the pharmacokinetics and bioavailability of cinnamic acid in mice.Methods:Extraction of cinnamon oil from cinnamon and determining content of cinnamic aldehyde: According to the method of determined volatile oil, a mass of 150g cinnamon powder and 1500mL water were mixed in a 3000mL flask. The mixture was reflux heated for 5 hr. Then, 8mL volatile oil (suspension containing cinnamon oil and water) was obtained and extracted with 8mL diethyl ether, separated out water. Six mL cinnamon oil was obtained after diethyl ether was distilled out. Cinnamic aldehyde is the main effectivecomposition (The first part of Chinese Pharmacopoeia 2000 commands that its content is not less 75%). Zero point two mL Cinnamic aldehyde comparision was diluted to 100 mL with methanol, 5mL dilution was diluted to 200mL with methanol again. So did cinnamon oil. The dilutions were filtrated with 0.46um filter membrane and measured with RP-HPLC.The conditions of chromatography: The column was Hypersil CI8(4.6 X 250 mm,5 um)(Dalian Elite Analytical Instrument Co., Ltd.).The mobile phase was methanol-acetonitrile-water-acetic acid (25:26:49:0.3). The flow rate was 1.0 mL /min.The UV detector was set at 270nm. The sensitivity was 1.0 AUFS.Pharmacokinetics: Cinnamic acid was dissolved in methamol and diluted to 2000 pig/mL. The solution was diluted to 500, 400, 250, 200, 100, 80, 40, 25, 20, 10, 5, 2, 1, 0.5, 0.4ug/mL with methanol.100uL per above dilution, 200jaL blank plasma and 600uL methanol was mixed thoroughly. All mixtures were centrifugalized (4000r/min) 1 Omin and filtrated, The upper solutions were measured with RP-HPLC. The linear equation was set up, the peak area (1.00X105) of cinnamic acid was dependent variable, the concentration was independent variable. High, medium and low three standard plasma solutions (295, 29.5, 2.95 jig/mL) was prepared as above same method. The quantitation limit, the detection limit, the recovery and the precision weremeasured as the method in the second part of Chinese pharmacopoeia 2000.Cinnamon oil was diluted into 5 times with soybean oil. Eighty four mice were forbidden food but free water for 24 hr. Then the gorge of these mice were instilled (ig.) with the dilution in 20 mL/kg dose. Zero point six mL mouse blood was gathered in heparin tube at 1, 3, 20, 60, 70, 85, 90, 100, 110, 120, 150, 160, 180, 210 min after ig.. All heparin tube were centrifugalized (3000r/min)10 min. Two-hundred uL plasma and 700 uL methamol were mixed thoroughly. The mixture was centrifugalized (4000r/min) 10 min and filtrated, the upper solutions were measured with RP-HPLC.Cinnamic acid was dissolved in 1,2-propanediol until saturation. Sixty four mice were injected in vein (iv.) the saturated solution in 157 mg/kg dose and 0.6 mL blood was gathered in heparin tube at 1, 3, 7, 10, 20, 30, 50, 70, 105, 120 min after iv.. Other 84 mice were ig. with the saturated solution in 450mg/kg dose after forbidden food but free water for 24 hr. Zero point six mL blood was gathered in heparin tube at 1, 5, 7, 8, 9, 10, 20, 30, 50, 75, 105, 120, 150, 180 min after ig.. All heparin tube were disposed as above same method.Results: The concentration of cinnamic aldehyde in cinnamon oil was 80%. The linear range was from 2.95 to 295(ig/mL with a slope of 0.2340, the correlation coefficient was 0.9992(n=ll). The average recovery and RSD in thethree plasma concentrations were 99.2%, 4.6%; 108%, 3.9%; 87.5%, 6.8% respectively. Within-day precision RSD was 6.8%, 4.0%, 4.6% respectively. Between-day precision RSD was 1.0%, 3.9%, 11% respectively. The dete...
|
PDF Full Text Request