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Objective To analyze the chemical compounds of Shenqi Dihuang decoction by the ultraperformance liquid chromatography coupled with linear quadrupole ion trap-orbitrap mass spectrometry (UPLC-LTQ-Orbitrap-MS). Methods Warters ACQUITY UPLC HSS T3 (2.1 mm ×100 mm, 1.8 μm) was used as chromatographic column with mobile phase: 0.1% formic acid water (A)-0.1% formic acid acetonitrile (B) with gradient elution, and flow rate was 0.3 ml/min. Electrospray ion source (ESI) and an electrostatic field orbital ion trap mass analyzer were adopted, which was used to collect mass spectrometry fragment information with positive and negative ion modes, by comparing with the relative retention time of the reference substance. In addition, the fragment information of the mass spectrum was used to identify the compounds. The accurate identification of the chemical components in Shenqi Dihuang decoction was confirmed with literature. Results The study found that UPLC-LTQ-Orbitrap-MS technology could be used to identify 62 chemical components, including 13 aromatic acids, 9 flavonoids, 8 saponins, and 5 aromatic amines, 3 keto acids, 2 phenols, 1 aromatic quinone and other ingredients in Shenqi Dihuang decoction. Conclusion The identification analysis method in this study was efficient and accurate, which could be applied to the identification and analysis of chemical components in Shenqi Dihuang decoction and provided the important experimental data for the research on the material basis and mechanism.
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Objective To collect the serum samples of patients with nonalcoholic fatty liver disease (NAFLD), and to investigate the changes in serum metabolic biomarkers before and after lifestyle intervention. Methods A total of 23 patients who were diagnosed with NAFLD in Department of Gastroenterology and Inpatient Department, Putuo District Central Hospital of Shanghai, from January 2019 to January 2020 were enrolled, and all patients received the intervention with aerobic exercise and equicaloric low-carbohydrate high-protein diet. A total of 13 healthy volunteers who underwent physical examination in Physical Examination Center were enrolled as control group. For the patients with NAFLD, basic information was collected before and after intervention, blood samples were collected twice to measure liver function, blood glucose, and blood lipids, and part of serum was used for serum metabolomics analysis. The serum samples were analyzed by ultra-performance liquid chromatography/tandem high-resolution mass spectrometry. The data collected were processed in Compound Discover, and then principal component analysis (PCA) and orthogonal partial least squares discriminant analysis were used to establish the profile of differentially expressed blood metabolites between patients and healthy people and perform the enrichment analysis of differentially expressed metabolic pathways. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Wilcoxon non-parametric test was used for comparison of non-normally distributed continuous data between two groups. Results After lifestyle intervention, the patients had significant reductions in body mass index ( P < 0.01), body weight ( P < 0.01), and serum biochemical parameters alkaline phosphatase, albumin, gamma-glutamyl transpeptidase, and alanine aminotransferase (all P < 0.05), as well as a significant reduction in total protein ( P < 0.01), while there were no significant improvements in cholinesterase, aspartate aminotransferase, and glucose. As for the four items for blood lipids, there was a significant reduction in triglyceride ( P < 0.01), while there were no significant improvements in high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and total cholesterol. The metabolomics analysis showed that 33 serum metabolites changed significantly after lifestyle intervention. In addition, PCA results showed that after intervention, the level of metabolites in patients tended to be normal. The signaling pathway analysis showed that exercise and diet mainly affected the pathways of bile acid, unsaturated fatty acid synthesis, and phenylalanine metabolism. Conclusion Lifestyle intervention can achieve varying degrees of reduction in the body weight of patients with NAFLD, improve serum biochemical parameters, and regulate the abnormal metabolic pathway in patients with NAFLD, which has important clinical value and significance for guiding clinicians to formulate reasonable diet and exercise strategies for patients with NAFLD and prevent the progression of NAFLD.
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Objective:To establish a method to determine vitamin B 6 and fluocinonide in compound vitamin B 6 gel.Methods:The contents of vitamin B6 and fluocinonide in the gel were determined by HPLC with a Lichrospher CN C 18column (250 mm ×4.6 mm, 5μm).The mobile phase was heptane sulfonate solution (adding 6.8 g potassium phosphate monobasic and 1.0 g heptane sulfonate into 1000 ml water) -methanol –triethylamine (35:65:0.2,adjusting pH to 3.2 with phosphoric acid).The detection wavelength was 238 nm and the column temperature was 25℃.The injection volume was 20 μl.Results: Vitamin B6 had a good linear relationship within the range of 15.0-300.0 μg· ml-1(r=1.000 0), and the average recovery was 99.65% (RSD=0.3, n=9).Fluocinonide had a good linear relationship within the range of 0.4-8.0 μg· ml-1 (r=0.9990), and the average recovery was 99.35% (RSD=0.85, n=9).Conclusion:The method is simple and reproducible, and the result is accurate.The method can be used for the deter-mination of the gel.
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Objective:To establish an HPLC method for the simultaneous determination of minoxidil and tretinoin in compound minoxidil gel. Methods:A Shim-Pack VP-ODS C18(250 mm × 4. 6 mm,5 μm)column was used with the mobile phase of methanol and 2% acetic acid solution by gradient eleution . The detection wavelength was respectively set at 280 nm and 350 nm,and the flow rate was 1. 0 ml·min -1 . The column temperature was 35℃ and the injection volume was 10 μl. Results:Minoxidil and tretinoin had a good linear relationship within the range of 4. 0- 240. 0 μg·ml-1 and 0. 05- 3. 00 μg·ml-1(r =0. 999 6 and 0. 999 3),respectively. The average recovery was 99. 56%(RSD = 0. 42% ,n = 9)and 99. 36%(RSD = 0. 50% , n = 9),respectively. Conclusion:The established method is simple,and suitable for the simultaneous determination of the two constituents in compound minoxidil gel.
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OBJECTIVE: To investigate the effect of interferon-? and verapamil on the reversal of multidrug-resistance(MDR) in breast cancer cell line.METHODS: Drug-sensitive MCF-7 and ADM-induced MDR phenotypic MCF-7/ADR were taken as in vitro experimental models,which were treated with interferon-? and verapamil in combination or alone,then the cell survival rate,IC50,drug-resistant times and reversal times of different experiment groups were measured by MTT test.The quantitive expression of P-170 on the surface of cells was detected by flow cytometry.RESULTS: Treated by interferon-? and verapamil in combination,the IC50 of adriamycin-resistance was lowered to 0.32 ?mol?L-1,superior to verapamil(1.23 ?mol?L-1) or interferon-?(2.29 ?mol?L-1) alone;the reversal times increased to 51.88 in combination group,superior to interferon-? and verapamil alone(7.25 and 13.49 respectively) and the expression of P-170 were lower than in groups treated with interferon-? and verapamil alone.CONCLUSION: The efficacy of interferon-? and verapamil exhibited a more remarkable efficacy on the reversal of multidrug-resistance(MDR) in breast cancer cell line when used in combination than alone.