ABSTRACT
Objective To establish a new quality control standard for danshen capsules. Methods The qualitative identification of danshen capsules was characterized by ultraviolet fluorescence and thin-layer chromatography( TLC ). The contents of tanshinoneⅡA,cryptotanshinone,salvianolic acid B,danshensu and protocatechuic aldehyde in danshen capsules were determined by high performance liquid chromatography(HPLC)on a C18 column. The flow rate was 1 mL·min-1,and the column temperature was 35 ℃. Results The HPLC linear ranges of tanshinone ⅡA,cryptotanshinone,salvianolic acid B, danshensu and protocatechuic aldehyde were 2. 046-40. 92 μg · mL-1 ,1. 482 25 -59. 29 μg · mL-1 ,1. 502 55 -60. 102 μg·mL-1 ,11. 49-459. 582 μg·mL-1 ,and 0. 617 4-24. 696 μg·mL-1 ,respectively,and r values were 0. 999 9. The average recoveries were 99. 66%(RSD of 0. 91%)for tanshinoneⅡA,99. 26%(RSD of 0. 88%)for cryptotanshinone,99. 09%(RSD of 0. 76%)for salvianolic acid B,100. 51%(RSD of 0. 62%)for danshensu,and 100. 62%(RSD of 0. 82%)for protocatechuic aldehyde,respectively. The contents of the tanshinoneⅡA,cryptotanshinone,salvianolic acid B,danshensu showed a certain high level in the 3 batches of danshen capsule samples,but protocatechuic aldehyde was low by comparison. Conclusion The HPLC method is proven to be sensitive,accurate,repeatable,and can be used for quality control of the danshen capsules.
ABSTRACT
Objective To investigate the expression of lipid rafts (LRs) and actin cytoskeleton (F-actin) in the peripheral blood B lymphocytes of patients with systemic lupus erythematosus (SLE).Methods Peripheral blood mononuclear cells (PBMCs) were separated by Ficoll-Hypaque.B lymphocytes were isolated by positive selection from PBMCs.Membrane staining for LRs was achieved with FITC-conjugated cholera toxin B (CTB).The level and distribution of LRs were studied by flow cytometry and confocal microscopy.Staining for F-actin was carried out with Rhodamine phalloidin.The expression of F-actin was analyzed by confocal microscopy.In an in vitro examination,the effect of Leflunomide on lipid rafts in B lymphocytes from SLE was analyzed.Disease carried out was measured using the SLE disease activity index (SLEDAI).Analysis of the enumerical data was performed using ANOVA or paired-samples t test.Correlation was examined by Pearson's rank correlation test.Results The number of CTB-binding lipid rafts in B cell from active SLE patients or from SLE patients in disease remission.who were treated with immunosuppressive drugs was higher than B cells from healthy controls [(59+4)％,(51±5)％,(33±4)％,F=9.21,P=0.001].Confocal microscopy revealed that in B cell from healthy controls,lipid raft was found to be small and uniformly distributed on the plasma membrane.F-actin was found mainly in the cortical region of the cells.This pattern was different from the pattern seen in B cells from patients with SLE,which presented with stronger staining and irregular large clustering of LRs,with a decrease in F-actin levels.In addition,the number of CTB-binding LRs in B cells from the active SLE patients was correlated significantly with the SLEDAI score (r=0.632,P=0.028).Furthermore,thein vitro results showed that leflunomide treatment reduced the number of CTB-binding LRs in B cell from SLE patients [(48±5)％ vs (39±5)％,t=2.29,P=0.048].Conclusion The altered expression of Lipid raft and F-actin can been seen in B lymphocytes in SLE,and modulation of LRs and F-actin expression may be a potential approach in the treatment of SLE.
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Object To study the pharmacokinetics of loganin in LIUWEI DIHUANG DECOCTION (LWDHD) in mice. Methods HPLC-UV detection was used to determine the loganin levels in biological samples. Results After ig LWDHD in mice, the plasma concentration-time course fitted well to one-compartment model with the 1st order absorption and with the following pharmacokinetic parameters; Ka= 0.04min -1, Ke=0.019 min -1, T (peak)=42.4 min, t 1/2ka=17.4 min, t 1/2ke=35.75 min. Conclusion The pharmacokinetic parameters of loganin obtained in the study may be attributed to the effect of other constituentsin the DECOCTION.