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China Pharmacy ; (12): 2243-2247, 2019.
Article in Chinese | WPRIM | ID: wpr-817166


OBJECTIVE: To establish the method for content determination of 6 kinds of triterpene acids such as haw acid, corosolic acid, betula acid, betulonic acid, oleanolic acid and ursolic acid in Tibetan medicine Rubus biflorus. METHODS: Pre-column devrivatization HPLC-FLD-APCI/MS method was adopted. 2-(7H-dibenzo[a,g]carbazol-7-yl) ethyl-4-methylbenzene- sulfonate was used as the pre-column derivatization reagent.Hypersil C18 column was used with the mobile phase consisted of 5% acetonitrile water solution-acetonitrile (gradient elution) at the flow rate of 1.0 mL/min. The excitation wavelength of fluorescence was 300 nm and the emission wavelength was 395 nm. The column temperature was 35 ℃, and sample size was 10 μL. Under atmospheric-pressure chemical ionization (APCI) source in positive-ion mode, pressure was 60 psi, the drying gas flow rate was 9 L/min, the dry gas temperature was 350 ℃, the gasification temperature was 450 ℃, and the capillary voltage was 3 500 V. RESULTS: The linear range of haw acid,corosolic acid,betula acid,betulonic acid,oleanolic acid and ursolic acid were 0.025-6.4 μg/mL(r≥0.999 6). The quantitative limits were 5.11, 4.78, 4.42, 4.22, 4.29, 4.51 ng/mL; and detection limits were 1.42, 1.27, 1.30, 1.28, 1.16, 1.22 ng/mL, respectively. RSD of precision test was less than 5%, stability and repeatability tests were all less than 2%(no betulonic acid detected). The recovery rates were 97.90%-100.55%(RSD=1.00%,n=6), 97.95%-102.95%(RSD=1.74%,n=6), 96.00%-101.20%(RSD=2.00%,n=6), 93.25%-104.20%(RSD=4.25%,n=6), 92.20%-103.30%(RSD=3.58%,n=6), 97.80%-103.50%(RSD=2.03%,n=6), respectively. CONCLUSIONS: The method is accurate, reliable and exclusive, and can be used for simultaneous determination of 6 kinds of triterpene acids in Tibetan medicine R. biflorus.

China Pharmacy ; (12): 179-182, 2018.
Article in Chinese | WPRIM | ID: wpr-704546


OBJECTIVE:To establish the quality standard of Tibetan medicine Rubus biflorus.METHODS:The qualitative identification was conducted from characters characteristics,microscopic characteristics,TLC.The contents of moisture,ash and extract were determined.HPLC method was adopted for content determination of rutin and hyperoside.The determination was performed on Diamonsil C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(18∶82,V/V) at the flow rate of 1.0 mL/min.The detection wavelength was set at 360 nm,column temperature was 30 ℃,and sample size was 10 μL.RESULTS:The surface of the medicinal material was grayish red to grey red brown,with longitudinal wrinkle grooves,and the peel was easy to peel off.The pith of the medicinal material was large,loose,sponge-like.The powder of the medicine was light yellow;wood fiber was bundle or scattered individually;bast fiber was thick.TLC spot of hyperoside was clear and well-separated.The contents of moisture,total ash,acid-insoluble ash,water-soluble extract and ethanol extract were 6.71%-7.55%,1.43%-1.67%,0.41%-0.48%,17.70%-19.31% and 13.76%-16.99%.The linear range was 3.13-50 μg/mL for rutin (r=0.999 2)and hyperoside (r=0.999 5).The limits of quantitation were 0.5,1.8 μg/mL,and the limits of detection were 0.2,0.9 μg/mL.RSDsof precision,stability and reproducibility tests were all lower than 2.0 % (n =6).The recoveries were 92.8 %-98.8 % (RSD =2.25 %,n=9),94.0%-98.6% (RSD=l.71%,n=9).CONCLUSIONS:The moisture content,total ash and acid insoluble ash content of medicinal materials is not more than 10.0%,3.0% and 0.6%,respectively.The total amount of water extract,alcohol extract,rutin and hyperoside is not less than 15.0%,12.0%,1.0 mg/g,respectively.Established standard can be used for quality control of Tibetan medicine R.biflorus.

China Pharmacy ; (12): 4699-4702, 2017.
Article in Chinese | WPRIM | ID: wpr-668658


OBJECTIVE:To establish the quality standard of Tibetan medicine Myricaria germanica. METHODS:M. germani-ca was identified in respects of properties,microscopic characteristics and TLC. The contents of moisture,ash and extract were de-termined. HPLC method was adopted for content determination of gallic acid. The determination was performed on Diamonsil C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(5:95,V/V)at the flow rate of 1.0 mL/min. The detection wavlengths were set at 270 nm. The column temperature was 30 ℃ and the sample size was 10 μL. RESULTS:M. ger-manica was cylindrical in shape,and there were many narrow strip alternate leaflets,which were brittle,easily broken,weak in smell,mild in flavor. Cork cell surface and lower epidermal cell surface showed polygon;most of xylon were bunched;there were many clusters of calcium oxalate. TLC spots were clear and well-separated. The contents of moisture,total ash,acid-insoluble ash, water-soluble extract and ethanol were 6.83%-8.12%,4.01%-5.01%,1.06%-1.98%,17.91%-22.65%,11.29%-15.51%. The linear range of gallic acid were 3.13-50 μg/mL(r=0.9997);RSDs of precision,stability and reproducibility tests were lower than 1.0%;the recoveries were 94.0%-100.8%(RSD=2.27%,n=9). CONCLUSIONS:Established standard can be used for quality evalua-tion of Tibetan medicine M. germanica.

Article in Chinese | WPRIM | ID: wpr-508135


Objective To study the effect of Tibetan MedicineManu-Xitanggranules on the expression of prostaglandin E2 and TNF-a on adjuvant arthritis (AA) rats.Methods A total of 60 rats were randomly divided into the control group, the model group, the Aspirin group, the low-, medium- and high- dose Manu-Xitang granules groups (each group with 10). Complete Freund's adjuvant method was used for the adjuvant arthritis model except the control group. All the groups started treatment at 8th day, and the treatment last 20 days. The low-, middle- high-dosage groups were treated with 2.0, 1.0, 0.5g/(kg body weight) Manu-Xitang granules, while the the Aspirin group with ASP (0.27 g/kg), control and medol groups with the equal Volume saline. The body weight, the swelling of primary side of arthritis index were observed. The levels of PGE2 and TNF-α were measured by ELISA.Results At 6th, 12th, 18th day, compared with the model group, the body weights in Aspirin group, the low-, middle-, high-dosage groups significantly increased (P<0.05 or P<0.01); at the 6th day, the foot swelling (0.40% ± 0.18%, 0.50% ± 0.25%, 0.55% ± 0.35%vs. 0.85% ± 0.15%) in the Aspirin group, middle-, and high-dosage groups significantly decreased (P<0.05 or P<0.01). And the PGE2 (0.66 ± 0.31vs. 0.39 ± 0.11) and TNF-a (0.72 ± 0.24vs. 0.50 ± 0.15) in the high- dose group were significantly lower than the model group (P<0.05).Conclusions The Tibetan medicineManu-Xitanggranules could treat AA through the changes of PGE2 and TNF-a.

Article in Chinese | WPRIM | ID: wpr-299770


Zuotai (gTso thal) is a typical representative of Tibetan medicines containing heavy metals, but there is still lack of modem safety evaluation data so far. In this study, acute toxicity test, sub-acute toxicity test, one-time administration mercury distribution experiment, long-term mercury accumulative toxicity experiment and preliminary study on clinical safety of Compound Dangzuo were conducted in the hope of obtain the medicinal safety data of Zuotai. In the acute toxicity test, half of KM mice given the lethal dose of Zuotai were not died or poisoned, and LD50 was not found. The maximum tolerated dose of Zuotai was 80 g x kg(-1). In the subacute toxicity test, Zuotai could reduce ALT, AST, Crea levels in serums under low dose (13.34 mg x kg(-1) x d(-1)) and medium dose (53.36 mg x kg(-1) x d(-1)), with significant difference under low dose, and increase the levels of ALT, AST, MDA, Crea in serums under high dose (2 000 mg x kg(-1) x d(-1)); besides, the levels of BUN and GSH in serums reduced with the increase in dose of Zuotai, indicating a significant dose-effect relationship. In the one-time administration distribution experiment, the content of mercury in rat kidney, liver and lung increased after the one-time administration with Zuotai, with a significant dose-dependent relationship in kidney. In the long-term mercury accumulative toxicity experiment, KM mice were administered with equivalent doses of Zuotai for 4.5 months and then stopped drug administration for 1.5 months. Since the 2.5th month, they showed significant mercury accumulation in kidney, which gradually reduced after drug withdrawal, without significant change in mercury content in liver, spleen and brain and ALT, AST, TBIL, BUN and Crea in serum. At the 4.5th month after drug administration, KM mice showed slight structural changes in kidney, liver and spleen tissues, and gradually recovered to normal after drug withdrawal. Besides, no significant difference in weight gain was found between the Zuotai group and the control group. According to the findings of the clinical safety study of Dangzuo, after subjects administered Dangzuo under clinical dose for one month, their serum biochemical indicators, blood routine indicators and urine routine indicators showed no significant adverse change. This study proved that traditional Tibetan medicine Zuotai was slightly toxic, with a better safety in clinical combined administration and no adverse effects on bodies under the clinical dose and clinical medication cycle. However, long-term high-dose administration of Zuotai may have a certain effect on kidney.

Adult , Animals , Female , Humans , Male , Mice , Middle Aged , Rats , Young Adult , Clinical Trials as Topic , Drugs, Chinese Herbal , Pharmacokinetics , Toxicity , Kidney , Liver , Medicine, Tibetan Traditional , Rats, Wistar