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OBJECTIVE：To study the effects and active ingr edients of Tibetan medicine Lamiophlomis rotata against rheumatoid arthritis （RA）. METHODS ：Fifty-six SD rats were randomly divided into normal control group （0.5％ sodium carboxymethylcellulose），model group （0.5％ sodium carboxymethylcellulose ），methotrexate group （positive control group ，3 mg/kg），L. rotata water extract low-dose ，medium-dose and high-dose groups （0.5，1，2 g/kg，by crude drug ），L. rotata total flavonoid group （200 mg/kg，by flavonoid extat ），with 8 rats in each group. Except for normal control group ，other groups were given Freund ’s complete adjuvant （FCA）on the rat ’s right hind footpad to induce adjuvant-induced arthritis model. The next day after injection of FCA ，rats in all groups were given relevant medicine intragastrically ，once a day （once every 3 days for methotrexate group ）for 30 days. At 15th and 30th day of administration ，the degree of paw swelling of left hind foot was measured，and the arthritis index ，spleen index were calculated. At the end of 30th day of administration ，the levels of TNF-α，IL-1β， IL-6 and IL- 10 in rat serum were determined by ELISA assay ，the thymus index and spleen index were calculated ，the pathological changes of the ankle joints were observed by HE staining. RESULTS ：Compared with normal control group ，degree of paw swelling，arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α，IL-1β， msxmX0146） IL-6 in serum at 30th day of administration were significantly com increased in model group （P＜0.01）；while the thymus indexyunbinji- and IL- 10 level at 30th day of administration were ang@swu.edu.cn significantly decreased（P＜0.01）；synovial cell proliferation and infiltration of articular cavity were observed in ankle joint. Compared with model group ，degree of paw swelling ，arthritis index at 15th and 30th day of administration as well as the spleen index and the levels of TNF-α，IL-1β，IL-6 in serum at 30th day of administration were significantly decreased in each medicine group （P＜0.01）；while the thymus index and IL- 10 level at 30th day of administration were significantly increased （P＜0.01）；the pathological changes of arthritis were significantly improved. Compared with L. rotata water extract high-dose group ，there were no significant differences in degree of paw swelling at 15th day of administration as well as the arthritis index ，spleen index ，levels of inflammatory cytokines and pathological changes of ankle joint in L. rotata total flavonoid group （P＞0.05）. CONCLUSIONS ：Tibetan medicine L. rotata shows well anti-RA activity ，and total flavonoids may be the active ingredients of its efficacy.
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Objective The HPLC-MS/MS is used to comprehensively monitor the feeding conditions of raw materials in Yuanhu analgesic capsules, and the content of the index components can be detected at the same time. Methods The Inertsil ODS-3 (4.6 mm×250 mm, 5 μm) was used with the column temperature 40 ℃, Flow phase: 0.1% formic acid-acetonitrile; the gradient elution program, active ingredients were separated by HPLC, and the Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode, and reactions ion monitoring mode (MRM) for quantitative analysis were selected. Results Through analysis and contrast of the medicinal materials, reference substance of primary mass spectrogram showed the same characteristic peak, and the proprietary Chinese medicine can be judged by prescription feeding process. The tetrahydroxene and imperatorin had a good linear relationship in 5.08×10-5-30.45×10-5μg (r=0.999 4), 5.02×10-5-30.09×10-5μg (r=0.999 2). Precision test were 0.99% and 1.14%, the recovery rate were 97.02%-99.66%, 97.62%-99.94%. Conclusions The method is simple, accurate and reliable, high sensitive and fast. It is suitable to monitor the feeding condition and quality of yuanhu analgesic capsules.
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Objective To develop a HPLC-MS/MS method for comprehensive monitoring and control of the raw material feeding (honeysuchle flowers extract and baikal skullcap root extract) , and simultaneous determination of chlorogenic acid and baicalin in Yinhuang capsules. Methods The separation was performed on an Inertsil ODS-3 (2.1 mm × 150 mm, 5 μm) analytical column with the mobile phase consisting of methanol- 10 mmol/L ammonium acetate solution by gradient elution program, and the column temperature was 40 ℃. Active ingredients were separated by HPLC, and the Electrospray Ionization Mass (API) source was applied and operated in the negative ion mode, and reactions ion monitoring mode (MRM) for quantitative analysis were selected. Results The samples and the mixed Extract have the same characteristic peak in MS and MS/MS. According to the prescription feeding process, the proprietary Chinese medicine wasdetermined. The results showed that the palladium residue of 6 batches samples were up to the standard by HPLC/MS/MS chromatographic peak areas. The calibration cruve of chlorogenic acid and baicalin were linear: 0.60-4.80 μg/ml (r = 0.9989),2.87-14.40 μg/ml (r = 0.9986), with the relative standard deviation of repeatability by 0.69% and 0.69% respectively, and the mean recovery rate were 95%-102%, 95%-103%, respectively. Conclusions The method was proven to be simple, accurate, reliable, high sensitive and can be used for determination and control of the raw material feeding (honeysuchle flowers extract and baikal skullcap root extract) and quality in Yinhuang capsules.
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Objective The monitoring of the feeding situation of active pharmaceutical ingredient in Shendan-Chenpi tablet by HPLC-MS/MS,and the content of sodium taurocholate and hesperidin.Methods Using ZORBAX Eclipse Plus C18(4.6 mm×250 mm, 5 μm)Column temperature 40 ℃, Mobile phase with acetonitrile-10 mmol/sodium acetate solution, using gradient elution program. Active ingredients were separated by HPLC, and the Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode, and reactions ion monitoring mode(MRM) for quantitative analysis were selected. Results The proprietary Chinese medicine is judged by the prescription feeding process, through analysis and contrast of the medicinal materials, reference substance of primary mass spectrogram, secondary mass spectrogram of peak. The aurocholic acid sodium and hesperidin had a good linear relationship in 0.242×10-2-1.45×10-2μg(r=0.996 0), 0.688×10-2-10.30×10-2μg(r=0.999 2), and the precision test were 0.78% and 1.56%, and the recovery rate were 102%-103%,96%-109%. Conclusions The method was simple, accurate and reliable, high sensitive and fast. The comprehensive monitoring was applied to the Shedan-Chenpi tablet in feeding analysis and quality.