Study On Chemical Constituents And Quality Of Ajuga Ovalifolia. Var. Calantha

Ajuga ovalifolia var.calantha is Lamiophlomis rotata (Benth.) Kudo plants Ajuga ovalifolia Bur.et Franch.var. calantha(Diels) C. Y. Wu et C. Chen whole hebra. It is one source of Tibetan medicine named tababa (sound translation).It can used to treat edema bloated yellow water,joint amass yellow water,Cancellation inflanation,injured wound,fracture contusion,physique sore and rheumatism joint sore, it also can used to stanch blood.This papers carry on maximum tolerance test and analgesic,effective parts of stanch bleeding filter on Ajuga ovalifolia var.calantha; and carry on extraction and separation of chemical composition on ajuga ovalifolia var.calantha; GC-MS analysis on ajuga ovalifolia var.calantha naphtha; assay three flavonoids ingredients of ajuga ovalifolia var.calantha; and Using high'performance liquid chromatography (HPLC) to study ajuga ovalifolia var.calantha fingerprint. The following is research results:1.Through the above pharmacology study on ajuga ovalifolia var.calantha of acute toxicity tests and analgesic hemostatic tests, it can be seen:the largest dosage of ajuga ovalifolia var.calantha water extrat and ethanol extract is 30g crude drug/kg weight, about 200 times for patients with clinical dosage, ajuga ovalifolia var.calantha water extrat and ethanol extract do not have obvious response, toxicology is proved safe.From the pharmacology study on ajuga ovalifolia var.calantha of acute toxicity tests and analgesic hemostatic tests, it can be seen:Ajuga ovalifolia var.calantha water extrat and ethanol extract have different degree of analgesia homeostatic function.2.Through extraction and separation,8 monomer (Ⅰ～Ⅷ) from ajuga ovalifolia var.calantha petroleum ether parts and ethyl acetate parts are got, and 7 compounds (Ⅱ～Ⅷ) structure are identified. Among them, compoundⅡis identified as usolic acid; compoundⅢis identified asβ-sistosterol; compoundⅣis identified as genkwanin; compoundⅤis identified as luteolin; compoundⅥis identified as apigenin; compoundⅦis identified as daucosterol; compoundⅧis identified asBreviflorasterone. These 7 compounds are got from ajuga ovalifolia var.calantha for the first time.3.GC-MS analysis on ajuga ovalifolia var.calantha naphtha. In the GC map of ajuga ovalifolia var.calantha naphtha, there are 34 peak, identified 29 compounds structure from MS analysis, its content is account for 98.12% of total naphtha. Among it,6 aliphatic olefin alcohols composition,39.63% of the amount,5 fatty acid composition, 49.42% of the amount,2 hemiterpene composition. The main composition of naphtha is 9,12,15-Octadecatrien-1-ol,[Z,Z,Z]-SS,the contents are 34.39%, n-Hexadecanoic acid is 42.79%.4.Building the methods to measure genkwanin and apigenin contents of ajuga ovalifolia var.calantha by using HPLC through methodology. Stationary phase:C18 chromatographic column; mobile phase:methanol-0.1% glacial acetic acid aqueous solution(80:20); velocity of flow:1.0ml/min; detection wavelength:269nm.Genkwanin standard curve equation is:A= 3093979x-45295 (r=0.9999).Sample size is between 0.1248μg～1.248μg, Genkwanin peak area has good linear relationship with sample size. RSD in the Precision tests,repeatability tests,recovery rate tests all are less than 2.0%, which were coincidence with stipulation.Building the methods to measure luteolin contents of ajuga ovalifolia var.calantha by using HPLC through methodology. Stationary phase:C18 chromatographic column; mobile phase:methanol-0.1% glacial acetic acid aqueous solution(70:30); velocity of flow:1.0ml/min; detection wavelength:350nm. Luteolin standard curve equation is:A= 3318415x-70716 (r=0.9996).Sample size is between 0.1264 u g～1.2640 u g, Luteolin peak area has good linear relationship with sample size. RSD in the Precision tests,repeatability tests,recovery rate tests all are less than 2.0%, which were coincidence with stipulation.Determinated 3 lot samples by building method. On the dried basis, genkwanin contents is 0.058%～0.073%,apigenin contents is 0.069%～0.085%,luteolin contents is 0.092～0.123%.5.Building the methods to measure fingerprint of ajuga ovalifolia var.calantha by using HPLC through luteolin as reference substance. C18 column as stationary phase; mobile phase:acetonitrile-0.1% glacial acetic acid aqueous solution gradient elution, gradient:15:85～35:65 (Omin～30min); 35:65～80:20 (30min～45min); 80:20～100: 0 (45min～50min).sample size is 10μl,detection wavelength:325nm, chromatogram recording time is 65 minute. The methods precision,repeatability are up to the requirement through methodology inspection. Building contrasting fingerprint of ajuga ovalifolia var.calantha by identifying 1,0 lots samples. Among 10 lots samples,13 characteristic peaks area sum is greater than 90%. So we can draw the conclusion that fingerprint has representativeness, it can provide the basis for controlling over ajuga ovalifolia var.calantha whole quality.