ABSTRACT
Objective::To investigate the effects of black bean juice with different stewing times on the appearance character and the content changes of effective components of Polygoni Multiflori Radix Praeparata. Method::HPLC was employed with Agilent ZORBAX Extend-C18 column (4.6 mm×250 mm, 5 μm), a gradient mobile phase of methanol (A)-water (B) was eluted (0-30 min, 5%-100%A; 30-40 min, 100%A), the flow rate was 1.0 mL·min-1, the injection volume was 10 μL and the column temperature was 35 ℃, detection wavelength was set at 280 nm. The contents of stilbene glycoside, emodin, emodin methyl ether, emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside in samples prepared at different processing times were simultaneously determined by HPLC. Result::The content of stilbene glycoside decreased gradually with the increase of stewing time, compared with 8 h, its content decreased by 76% at 64 h. The contents of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside increased first, and then decreased, reaching the highest value at 24 h, and then decreased to the level similar to the content of 8 h after 40 h, and then fluctuated slightly. The contents of emodin and emodin methyl ether increased first, and then decreased, reached the maximum when stewed for 32 h, then decreased slowly and tended to be stable. Conclusion::The stewing time has significant influence on the content of various components in Polygoni Multiflori Radix Praeparata, and the changing trend is different, the processing time of Polygoni Multiflori Radix Praeparata shall be standardized. At the same time, it is not sufficient to take stilbene glycoside and anthraquinones as the indicator ingredients for this decoction pieces, the quality control indicators such as polysaccharides shall be considered.
ABSTRACT
Objective: To optimize the extract process of anthraquinones from the Rhei Radix et Rhizoma (RRR). Methods: The contents of eight bound anthraquinones (Bas) and five free anthraquinones (Fas), a total of 13 original anthraquinones, and total anthraquinones (Tas) in RRR were determined by HPLC. L9(34) orthogonal table was used, with ethanol concentration, solvent ratio, extract time, and number of extract as factors and extract quantity of Tas, five Fas, and eight Bas as the investigation index, respectively, the extract processes of Tas, Fas, and Bas from RRR were optimized. The optimized processes were verified by magnification test. Results: The optimum extract process of Tas and Fas was as follows: five times of 75% ethanol, extracting five times by reflux, 30 min for each time. Using this process, the extraction rate for both original anthraquinones and Tas was about 90%, and Fas extraction rate was over 160%. The optimum extract process of Bas was as follows: five times of 95% ethanol, extracting three times by reflux, 60 min for each time. Using this process, original anthraquinones extraction rate was close to 90%, and the one of Bas was over 80%. Conclusion: The optimized process is simple, stable, feasible, and repeatable, and can be used for the extraction of Tas, Fas, and Bas from RRR, respectively.
ABSTRACT
Diabetic nephropathy (DN) is the leading cause of end-stage failure of the kidney, but the efficacy of currently available strategies for the prevention of DN remains unsatisfactory. In this study, we investigated the effects of free anthraquinones (FARs) extract, which was extracted from the rhubarb and purified by macroporous resin DM130 with gradient mixtures of ethanol/water as the lelution solvents, in high glucose-cultured glomerular mesangial cells (MCs). The cell proliferation was determined by CCK-8 assay, the levels of TGF-beta1, CTGF, ColIV and FN proteins in the supernatant of MCs were measured by ELISA assays, and the mRNA levels of these four genes were detected by RT-PCR. The results showed that the increased proliferation of MCs, the mRNA levels and protein expression of TGF-beta1, CTGF, ColIV and FN induced by high glucose were inhibited after the treatment with the FARs extract. This indicated that FARs extract could inhibit cell proliferation and the expression of main extracellular matrix induced by high glucose in MCs. The FARs extract exhibited potential values for prophylaxis and therapy of DN.
Subject(s)
Anthraquinones , Cell Proliferation , Diabetic Nephropathies , Enzyme-Linked Immunosorbent Assay , Extracellular Matrix , Glucose , Kidney , Mesangial Cells , Rheum , RNA, Messenger , Sincalide , Solvents , Transforming Growth Factor beta1ABSTRACT
Objective: To study the in vitro transdermal absorption characteristics of the free anthraquinones of rhubarb in Dasuan Xiaohuang Pasta (DXP) and investigate the effect of various excipients (vinegar, ginger juice, and vegetable oil) and different compatibilities on rhubarb transdermal characteristics. Methods: Taking rhubarb free anthraquinones, such as aloe emodin, rhein, emodin, chrysophanol, and physcion as indexes, the studies were carried out by in vitro transdermal absorption. Results: The transdermal absorption of free anthraquinones of rhubarb with various excipients and different compatibilities were all zero-order kinetics; Using vinegar as the excipient, the 8 h accumulation permeation amount and transdermal rate of every ingredient were the largest, followed by ginger juice and vegetable oil; In the different compatibility tests, the 8 h accumulation permeation amount and transdermal rate of each ingredient in anagraph were the largest, followed by rhubarb-sodium sulfate and rhubarb-garlic, and the single rhubarb was the lowest. Conclusion: DXP using vinegar shows the best, followed by ginger juice, while vegetable oil shows the worst. Garlic and sodium sulfate show the improvement on transdermal absorption of free anthraquinones of rhubarb, and sodium sulfate shows better effect than garlic.
ABSTRACT
Objective To establish a method for the content determination of the free anthraquinones,garlic acid,berberine hydrochloride and palmatine hydrochloride in Shuangbai San,and to provide a reference for the quality control and chemical analysis of Shuangbai San.Methods HPLC with Eclipse XDB C18 column was used for the determination of the free anthraquinones.The chromatographic conditions were as follows:methanol-0.1 %phosphoric acid as a mobile phase in gradient mode and detection wavelength at 254 nm,flow rate at 1 mL?min-1,and column temperature at 30 ℃.The chromatographic conditions for the determination of gallic acid were as follows:Nucleodur C18 Gravity column,methanol-0.1 %phosphoric acid solution(2 ∶98) as mobone phase,detection wavelength at 273 nm,flow rate at 1 mL?min-1,and column temperature of 40 ℃.The chromatographic conditions for the determination of berbertine hydrochloride and palmatine hydrochloride were as follows.Nucleodur C18 Gravity,acetonitrile-0.1 %phosphoric acid(every 100 mL solution containing sodium dodecyl sulfonate 0.1 g)(39 ∶61) as mobile phase,detection wavelength at 345 nm,flow rate at 1 mL?min-1,and column temperature being 40 ℃.Results The calibration curves were linear(r≥0.999 7),and the average recoveries were all between 95 %～105 %.The total content of free anthraquinones was 5.1172～5.4933 mg?g-1,the content of gallic acid was 1.3376～2.0673 mg?g-1,and that of berbertine hydrochloride and palmatine hydrochloride were 1.2812～1.6855 mg?g-1.Conclusion This method is reliable and simple,which can provide a reference for the quality control and chemical analysis of Shuangbai San.