A simple method has been proposed for the determination of clozapine (CLZ) and chlorpromazine (CPZ) in human urine by dispersive liquid-liquid microextraction (DLLME) in combination with high-performance liquid chromatography-ultraviolet detector (HPLC-UV). All important variables influencing the extraction efficiency, such as pH, types of the extraction solvent and the disperser solvent and their volume, ionic strength and centrifugation time were investigated and optimized. Under the optimal conditions, the limit of detection (LODs) and quantification (LOQs) of the method were 13 and 39 ng/mL for CLZ, and 2 and 6 ng/mL for CPZ, respectively. The relative standard deviations (RSDs) of the targets were less than 5.1% (C=0.100 μg/mL, n=9). Good linear behaviors over the tested concentration ranges were obtained with the values of R (2)>0.999 for the targets. The absolute extraction efficiencies of CLZ and CPZ from the spiked blank urine samples were 98.3% and 97.8%, respectively. The applicability of the technique was validated by analyzing urine samples and the mean recoveries for spiked urine samples ranged from 93.3% to 105.0%. The method was successfully applied for the determination of CLZ and CPZ in real human urine.

OBJECTIVETo study the chemical constituents of Macrothelypteris viridifrons and their anti-proliferative effects on tumor cell.

METHODThe compounds were isolated by column chromatography with silica gel, C18 reverse-phase silica gel, sephadex LH-20, and their structures were elucidated on the basis of physiochemical propertities and spectral analysis. The antitumor activities of all compounds were tested with MOLT4, Hep G2, A-549, MCF-7, HT-29, PC-3 tumor cell lines.

RESULTFive compounds were isolated and identified as protoapigenone (1), protoapigenin (2), protoapigenin-4'-O-beta-D-glucopyanoside (3), 5,7-dihydroxy-2-(1,2-isopropyldioxy-4-oxo-cyclohex-5-enyl) -chromen-4-one (4), 5,7-dihydroxy-2-(1-hydroxy-2,6-dimethoxy-cyclohex-4-oxo) -chromen-4-one (5), respectively.

CONCLUSIONAll compounds were obtained from this plant for the first time. Compounds 1, 4 and 5 showed strong anti-proliferative effects on six tumor cells, which were in concentration-dependent manner.

Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ferns ; chemistry ; drug effects ; Flavonoids ; chemistry ; pharmacology ; HT29 Cells ; Hep G2 Cells ; Humans

A simple method has been proposed for the determination of clozapine (CLZ) and chlorpromazine (CPZ) in human urine by dispersive liquid-liquid microextraction (DLLME) in combination with high-performance liquid chromatography-ultraviolet detector (HPLC-UV).All important variables influencing the extraction efficiency,such as pH,types of the extraction solvent and the disperser solvent and their volume,ionic strength and centrifugation time were investigated and optimized.Under the optimal conditions,the limit of detection (LODs) and quantification (LOQs) of the method were 13 and 39 ng/mL for CLZ,and 2 and 6 ng/mL for CPZ,respectively The relative standard deviations (RSDs) of the targets were less than 5.1％ (C=0.100 μg/mL,n=9).Good linear behaviors over the tested concentration ranges were obtained with the values of R2＞0.999 for the targets.The absolute extraction efficiencies of CLZ and CPZ from the spiked blank urine samples were 98.3％ and 97.8％,respectively.The applicability of the technique was validated by analyzing urine samples and the mean recoveries for spiked urine samples ranged from 93.3％ to 105.0％.The method was successfully applied for the determination of CLZ and CPZ in real human urine.

OBJECTIVE： To optimize the extraction technology of Shengmaiyin polysaccharide， and to investigate the regulation effects of Shengmaiyin and its polysaccharide on intestinal function of spleen deficiency model rats. METHODS： The contents of polysaccharide were determined by phenol-sulfuric acid method， and the extraction rate of polysaccharide was calculated. Using extraction rate of Shengmaiyin polysaccharide as investigation index， singel factor and orthogonal tests were used to optimize material-liquid ratio， extraction time， extraction temperature and extraction times of Shengmaiyin polysaccharide. Validation test was also conducted. Totally 80 male SD rats were randomly divided into blank group， model group， Shengmaiyin low-dose， medium-dose and high-dose groups （350， 700， 1 400 g/L， by crude drug）， Shengmaiyin polysaccharide low-dose， medium-dose and high-dose groups （24.5， 49， 98 g/L， by crude drug）， with 10 rats in each group. Except for blank group， other groups were given Rheum palmatum water decoction 10 mL/kg to induce spleen deficiency model， once a day， for consecutive 15 d. Since the 16th day， blank group and model group were given isovolumic water intragastrically， while other groups were given corresponding drugs， once a day， for consecutive 10 d. The general status of rats and body weights were recorded in each group. The serum contents of D-xylose， gastrin （GAS） and vasoactive intestinal peptide （VIP） were detected by phloroglucinol method or ELISA. RESULTS： The optimal extraction technology of Shengmaiyin polysaccharide was material-liquid ratio 1 ∶ 10（g/mL）， extraction time 45 min， extraction temperature 80 ℃， extracting for 1 time. Results of validation test showed that extraction rates of the polysaccharide in 3 times were 7.43％， 7.64％， 7.80％ （RSD＝1.01％， n＝3）. After modeling， except for blank group， other groups suffered from loose stools， thin body and reduced food intake， and the body weight and serum level of D-xylose were decreased significantly compared with blank group （P＜0.01）. After last medication， above symptoms of administration groups were improved to different extents. Except for model group， body weight and serum contents of D-xylose in other groups were increased significantly than those before modeling or before medication （P＜0.05 or P＜0.01）. Compared with blank group， body weight and serum content of GAS were decreased significantly in model group， while serum content of VIP was increased significantly （P＜0.01）. Compared with model group， body weight of Shengmaiyin medium-dose group and Shengmaiyin polysaccharide low-dose and high-dose groups， serum contents of D-xylose and GAS in Shengmaiyin medium-dose and high-dose groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were increased significantly， while serum contents of VIP in Shengmaiyin groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： The optimized extraction technology of Shengmaiyin polysaccharide is stable and feasible. Shengmaiyin and its polysaccharide contribute to the recovery of intestinal function of spleen deficiency model rat， the effects of which may be associated with the secretion regulation of GAS and VIP.

Traditional Chinese herbs (TCH) are currently gaining attention in disease prevention and health care plans. However, their general bitter taste hinders their use. Despite the development of a variety of taste evaluation methods, it is still a major challenge to establish a quantitative detection technique that is objective, authentic and sensitive. Based on the two-bottle preference test (TBP), we proposed a novel quantitative strategy using a standardized animal test and a unified quantitative benchmark. To reduce the difference of results, the methodology of TBP was optimized. The relationship between the concentration of quinine and animal preference index (PI) was obtained. Then the PI of TCH was measured through TBP, and bitterness results were converted into a unified numerical system using the relationship of concentration and PI. To verify the authenticity and sensitivity of quantified results, human sensory testing and electronic tongue testing were applied. The quantified results showed a good discrimination ability. For example, the bitterness of Coptidis Rhizoma was equal to 0.0579 mg/mL quinine, and Nelumbinis Folium was equal to 0.0001 mg/mL. The validation results proved that the new assessment method for TCH was objective and reliable. In conclusion, this study provides an option for the quantification of bitterness and the evaluation of taste masking effects.