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OBJECTIVE:To establish a method for content determination of steroidal saponin in Allium sativum. METHODS:Pre-column derivatization of steroidal saponin was performed by using the derivatization agent of nitro-benzoic acid-chlorine. And HPLC was conducted to determine the content of steroidal saponin. The column was Shimadzu VP-ODS with mobile phase of aceto-nitrile-water mixed solution(80:20,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 254 nm,the column temper-ature was 25 ℃,and the injection volume of 20 μl. RESULTS:The linear rang of sarsasapogenin was 0-1.25 mg/ml(r=0.999 0);RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 93.1%-96.8%(RSD=1.56%,n=6). CON-CLUSIONS:The method is simple,stable with good separation,and can be use for the content determination of steroidal saponin in A. sativum.
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Aim: To observe the effects of sarsasapogenin ( SAR) on osteoblasts and osteoclasts cultured in vitro. Methods: Colonal murine calvarial osteoblast-like cell line MC3T3-E1 cells were cultured in vitro. MTT,p-nitropheneye phosphate and tinctorial method of alizarin Bordeaux were used to investigate the effects of SAR on the proliferation, ALP expression, and mineralization tuberculation of MC3T3-E1 cells. Mature osteoclasts were i-solated from the long bone of one-day rat. Meanwhile, marrow cells of mouse bone were cultured with induction of 1,25( OH)_2VitD_3. During the culturing of osteoclasts or marrow cells, SAR of different concentrations was added into the medium. The number of osteoclasts was recognized as tartrate resistant acid phosphatase( TRAP) ( +) multinucleate cells and the resorption lacuna on bone slice were examined with toluidine blue staining. Results: Comparing with the control group, SAR (0.01, 0. 1, 1μg/mL) significanthy increased the proliferation of MC3T3-E1 cells (P <0. 05, P <0. 01). There was no significant difference in the expression of ALP in early pro-liferating MC3T3-E1 cells exposed to SAR of 0.01,0. 1, 1μg/mL, but in the differentiation phase MC3T3-E1 cells, SAR improved ALP activity very significantly if compared with the control group, of which SAR of 1 μg/mL had the most promotion effect(P <0. 01). In addition, compared to the control group, there were, to various ex-tents, increased in the number of mineral nodes in MC3T3-E1 cells after 15day incubation with SAR of different conentrations. Furthermore, no obvious effects of 0.01-1μg/mL SAR on mature osteoclast were observed. But typical osteoclasts were formed when marrow cells were cultured with the induction of 1,25(OH)_2D_3 in medium for 7 days while little or no osteoclasts were induced from marrow cells in the presence of SAR. Conclusion: The results suggest that SAR can effectively promote the proliferation, differentiation and mineralization of osteoblasts cultured in vitro. Besides, SAR can inhibit the generation of osteoclasts from marrow cells.
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OBJECTIVE:To establish a TLCS method for the determination of Sarsa sa pogenin in Kangbingdu oral liquid.METHODS:TLCS was used with benzol-acetone(9 ∶1)as developing agent Sawtooth scanning was at ?S=443nm,?R=600nm RESULTS :The linear range was 1 028?g～3 624?g(r=0 9 995) The average recovery was 100.89%,and the RSD was 3 36%(n=5) The stability and precision were go od CONCLUSION:TLCS method can be used for the determination of Sarsasapogenin in Kangbingdu oral liquid
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Object To show that ELSD is an excellent detector for the detection of chemical compounds devoid of chromophore, such as sarsasapogenin Methods Sarsasapogenin in crude Anemar rhena asphodeloides Bunge and its preparation was determinated by RP HPLC ELSD Results The well separated chromatographic peaks show linearity with recovery of the added sample of 100 5% in crude medicinal material and 91 38% in its preparation, r= 0 999 0 Conclusion The method was advanced, reliable, simple and can be used for quality control of crude A asphodeloides and its preparations
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OBJECTIVE:To determine Sarsasapogenin in Jujube seed concentrated pills by RP-HPLC-ELSD.METHODS:Separation of Sarsasapogenin was performed on Zorbax C18 column with methanol-water (90:10)as a mobile phase at a flow rate of 1.0mL?min-1.The temperature of the drift tube was 85℃and the air flow-rate was at 1.71mL?min-1.RESULTS:The linear range of Sarsasapogenin was 0.112 7～0.676 2mg?mL-1(r=0.998 3).The average recovery was 99.83%(RSD=0.93%).CONCLUSION:The method is simple,accurate and reproducible,and suitable for the quality control of Jujube seed concentrated pills.