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To analyze the content change of the nephrotoxic substances, aristolochic acid derivates [AAs] in the roots of Aristolochia debilis and the products generated from the solid-state fermentation of six different medicinal fungi by HPLC-ESI-TOF-MS, the chromatographic separation was carried out on C18 column at 30 degree C with the DAD detector. The elution was performed using the mobile phase of acetonitrile [A] and 0.2% acetic acid [B]. Several new peaks were found in the scale of 0-20 min elution of HPLC diagram in the fermentation products. The ESI-MS detection [negative ion mode] was carried out by post-column flow splitting method following the automatic injection. Seven AAs in the fermentation products and A. debilis were deduced, which were recognized as AAIa or IIIa [1], AAVIa [2], AAIVa, Va, VIIa or VIIIa [3]; AAII [4]; AAIII [5]; AAI [6]; AAIV or VII [7]. The areas of almost all these seven components existing originally in the corresponding crude drug decreased after the fermentation process, suggesting that fermentation is an effective way of lowering the nephrotoxicity induced by AAs in Chinese medicines similar with A. debilis. In addition, Optimized HPLC-MS method is helpful to AAs content identification
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Objective To prepare and purify the polysaccharides in Shuanqian fungal substance (FS);To study its monosaccharide composition and property. Methods Strychnos nux-vomica was under a thirty-day solid fermentation through Trametes cinnabarina. Then crude polysaccharide was got after the process of drying, smashing, water boiling, condensing, decoloring, removing protein and ethanol precipitation. The content of polysaccharides in Shuanqian FS was determined by phenol-sulfuric acid method. Dialysis and gel chromatography method was used to purify the crude polysaccharide. The purified composition was analyzed by acid hydrolysis and thin layer chromatography. Results The content of crude polysaccharide was 47.68%and yield was 10.52%. It showed that polysaccharide in Shuanqian FS contained glucose and galactose, but no nucleic acid and protein. Conclusion Polysaccharide in Shuanqian FS was an offwhite powdery heteropolysaccharide, which contained glucose and galactose.
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Objective To investigate the content change of aconitine in Radix Aconiti Kusnezoffii in different boiling time. Methods The chromatographic procedure was carried out with methanol-water-chloroform-diethylamide (70∶30∶3∶0.2) as mobile phase at a flow rate of 1.0 mL/min, the column temperature at 30 ℃, and UV detector at 240 nm. Results There was a good linear relationship of aconitine within 0.08-0.8 μg, r=0.999 8 (n=6), and the average recovery rate was 97.16%. The content of aconitine increased gradually in 0-2 h, then gradually decreased. And the reduction was evident particularly during 2-4 hours. Conclusion HPLC method is rapid, simple, accurate, reproducible and reliable for determining aconitine. The content of aconitine can be decreased effectively by boiling 4 hours.
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Objective To optimize the extraction technology of alginic acid in Laminaria japonica Aresch. Methods The influence of the amount of solvent, the extraction time and temperature (3 factors) on the alginic acid content and yield was investigated by orthogonal test L9(34), and carried on to inspect the methodology. Result The optimum extraction condition was A1B1C2, that was adding 30 times solvent (1%Na2CO3), whisk extracting for one hour at 60 ℃. Conclusion The temperature is the most important factor. The content of alginic acid decrease obviously at above 60 ℃. The optimum extraction technology is reasonable.
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Objective:To optimize the extraction procedure of total alkaloid in Radix Aconiti.Methods:The orthogonal design(L9(34) ) was carried out for the optimum extraction conditions guided by yield of the extract and the content of aconitine,hypaconitine and mesaconitine determined with RP-HPLC method,the mobile phase was MeOH-H2O2-CHCl3-DEA(70∶30∶ 2∶0.1).Results:The best extraction process was as follows:moisten with ammonia test solution,then leach for 24 hours with 15 times of ether.Conclusion:The optimum extraction methods are rational.