Dispersive liquid-liquid microextraction combined with high-performance liquid chromatography for the determination of clozapine and chlorpromazine in urine

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.     

翅柄铁线蕨治疗小鼠实验性良性前列腺增生有效部位的筛选

目的:研究翅柄铁线蕨对良性前列腺增生（BPH）模型小鼠的治疗作用,并筛选其有效部位。方法:皮下注射丙酸睾酮连续21d复制小鼠BPH模型,同时给药组分别灌胃给予翅柄铁线蕨总提取物及不同极性部位（石油醚、乙酸乙酯、正丁醇）(600mg·kg^-1·d^-1)。于末次给药后禁食,于次日剥离前列腺组织,称取前列腺湿质量,计算前列腺指数并进行病理学观察。结果:总提取物和乙酸乙酯部位组前列腺指数明显减小（P<0.05）,各用药组前列腺组织病理学变化均有不同程度的改善,其中乙酸乙酯部位组对BPH小鼠前列腺病理学改善效果最显著。结论:翅柄铁线蕨对BPH有一定的疗效,其中乙酸乙酯部位效果最好,初步确定为翅柄铁线蕨治疗BPH的有效部位。     

普通针毛蕨质量标准的研究

目的 研究普通针毛蕨的质量标准.方法 运用显微鉴定、薄层色谱、紫外可见分光光度法、高效液相色谱等技术对普通针毛蕨进行研究,对其水分、总灰分、酸不溶灰分、总黄酮、原芹菜素含量进行测定.结果 参照《中国药典》2005年版(Ⅰ部),普通针毛蕨生药中水分不得超过15.00％,总灰分不得超过8.31％,酸不溶性灰分不得超过3.90％,t检验显示,上述指标,因产地不同,无显著性差异(P＞0.05);生药中原芹菜素的含量不得低于0.71％,总黄酮含量不得低于28.21％.因药材产地不同,其总黄酮、原芹菜素含量有显著性差异(0.01＜P≤0.05).结论 实验结果为建立普通针毛蕨药材的质量标准提供了依据,方法简单易行.     

翠绿针毛蕨中特殊B环结构的黄酮及其抑制 肿瘤细胞增殖作用的研究

目的:研究翠绿针毛蕨Macrothelypteris viridifyons的化学成分及其抑制肿瘤细胞增殖作用.方法:利用硅胶、C18反相硅胶、Sephadex LH-20凝胶等色谱技术对其提取物进行分离纯化,根据化合物的理化性质和波谱数据进行结构鉴定,以MTT法测试各化合物对MOLT-4,Hep G2,A-549,MCF-7,HT-29,PC-3肿瘤细胞增殖的抑制作用.结果:分离得到5个具有特殊B环结构的黄酮类化合物,分别鉴定为:原芹菜素(1),4-羟基-原芹菜素(2),4'-羟基-原芹菜素4'-O-β-D-葡萄糖苷(3),5,7-二羟基-2-(1,2-异丙二氧基-4-酮-环己-5-烯)-色原酮(4),5,7-二羟基-2-(1-羟基-2,6-二甲氧基-环己-4-酮)-色原酮(5).结论:所有化合物均首次从翠绿针毛蕨中分离得到,化合物1,4和5对6种人肿瘤细胞增殖有明显的抑制作用,且呈现出良好的剂量依赖性.     

梯度反相高效液相色谱法测定普通针毛蕨地下部分Protoapigenone含量

目的 建立测定普通针毛蕨地下部分中Protoapigenone的梯度洗脱反相高效液相色谱法。方法 采用Eurospher C18色谱柱（250 mm× 4.0 mm,5 μm）,甲醇 水（0.1%磷酸,pH值＝3.0）为流动相,梯度洗脱,检测波长250 nm,柱温25 ℃。结果 Protoapigenone在7.5～480.0 mg&#8226;mL-1范围里线性关系良好,普通针毛蕨地下部分中Protoapigenone含量为1.14%,RSD为3.5%。结论 该法稳定、简便,可用于普通针毛蕨药材的质量控制。     

Inhibitory effect of diosgenin on experimentally induced benign prostatic hyperplasia in rats

This study investigated the effect of diosgenin, a natural sapogenin possessing various pharmacological activities, on benign prostatic hyperplasia (BPH) in rats and the possible mechanisms. BPH was established in the castrated rats by subcutaneous injection of testosterone propionate. Animals were randomly divided into four groups (n=10 each): model group (0.5% sodium carboxymethyl cellulose); positive control group (3 mg/kg finasteride); two diosgenin groups (50 and 100 mg/kg). The drugs were intragastricaly given in each group for consecutive 3 weeks. Another 10 rats with no testicles cut off served as negative controls and they were subcutaneously injected with 0.1 mL olive oil per day and then treated with 0.5% sodium carboxymethylcellulose. After 3-week administration, the prostate index and serum PSA level were determined, and histopathological examination was carried out. The levels of MDA, SOD and GPx in prostates were also measured. Additionally, the expression of Bcl-2, Bax and p53 was examined using Western blotting. The results showed that the prostate index and serum PSA level were significantly decreased, and the pathological changes of the prostate gland were greatly improved in diosgenin groups as compared with the model group. Elevated activities of SOD and GPx, and reduced MDA level were also observed in diosgenin-treated rats. In addition, the expression of Bcl-2 in prostates was down-regulated, whereas that of Bax and p53 was up-regulated in diosgenin-treated rats. These results indicated that diosgenin was effective in inhibiting testosterone propionate-induced prostate enlargement and may be a candidate agent for the treatment of BPH.     

Preparation of Magnetic Molecularly Imprinted Polymers for Selective Isolation and Determination of Kaempferol and Protoapigenone in Macrothelypteris torresiana

Novel uniform-sized magnetic molecularly imprinted polymers （MMIPs） were synthe- sized for selective recognition of active antitumor ingredients of kaempferol （KMF） and protoapi- genone （PA） in Macrothelypteris torresiana （M. torresiana） by surface molecular imprinting tech- nique in this study. Super paramagnetic core-sheU nanoparticles （γ-MPS-SiO2@Fe3O4） were used as seeds, KMF as template molecule, acrylamide （AM） as functional monomer, and N, N＇-methylene bisacrylamide （BisAM） as cross-linker. The prepared MMIPs were characterized by X-ray diffraction （XRD）, Fourier transform infrared spectrum fiT/R）, transmission electron microscopy （TEM） and thermo-gravimetric analysis （TGA）, respectively. The recognition capacity of MMIPs was 2.436 times of non-imprinted polymers. The adsorption results based on kinetics and isotherm analysis were in accordance with the pseudo-second-order model （R2=0.9980） and the Langmuir adsorption model （R2=0.9944）. The value of E （6.742 kJ/mol） calculated from the Dubinin-Radushkevich isotherm model suggested that the physical adsorption via hydrogen-bonding might be predominant. The Scatchard plot showed a single line （R2=0.9172） and demonstrated the homogeneous recognition sites on MMIPs for KMF. The magnetic solid phase extraction （MSPE） based on MMIPs as sorbent was established for fast and selective enrichment of KMF and its structural analogue PA from the crude extract of M. torresiana and then KMF and PA were detected by HPLC-UV. The established method showed good performance and satisfactory results for real sample analysis. It also showed the feasi- bility of MMIPs for selective recognition of active structural analogues from complex herbal extracts.     

针毛蕨的化学成分及其体内外抗肿瘤活性研究

 目的 研究针毛蕨（Macrothelypteris oligophlebia化学成分及其体内外抗肿瘤活性。方法 采用各种色谱技术进行分离纯化,根据理化性质和波谱数据进行结构鉴定,以MTT法测试各化合物对肿瘤细胞的体外抑制作用,并观察针毛蕨提取物对S₁₈₀肉瘤小鼠肿瘤生长的影响。结果 分离得到7个黄酮类化合物,分别鉴定为：荚果蕨素（1、柚皮素（2、柚皮素-5-O-β-D-吡喃葡萄糖苷（3、原芹菜素（4、去甲氧基荚果蕨素（5、芹菜素（6和原芹菜素-4′-O-β-D-葡萄糖苷（7。结论 所有化合物均首次从针毛蕨中分离得到;化合物4对肿瘤细胞增殖有显著的抑制作用,且呈现出良好的剂量依赖性;体内抗肿瘤试验显示针毛蕨提取物对S₁₈₀肉瘤小鼠肿瘤的生长有明显的抑制作用。     

翠绿针毛蕨中黄酮类化学成分的研究

目的 研究翠绿针毛蕨(Macrothelypteris viridifrons)中的黄酮类化学成分.方法 利用硅胶、C18反相硅胶、sephadexLH-20凝胶等色谱技术对其提取物进行分离纯化,根据化合物的理化性质和光谱数据进行结构鉴定.结果 分离得到9个黄酮类化合物,分别为:山柰酚-3-O-β-D-葡萄糖苷(1),...