棘白菌素B母核关键杂质的制备与鉴定

采用制备液相色谱技术对棘白菌素B母核粗品中的关键杂质进行分离纯化,并通过质谱和核磁共振波谱分析鉴定杂质结构。制备的2个棘白菌素B母核杂质(化合物M和N)与棘白菌素B母核具有相同的主体结构,分别为母核F位和E位氨基酸残基的去甲基产物,为阿尼芬净的杂质研究和质量控制提供了技术支持。     

米卡芬净的加压毛细管电色谱法测定

建立了加压毛细管电色谱法测定抗真菌药米卡芬净。采用EP-100-20/45-3-C18-BP反相毛细管色谱柱,以乙腈∶2.5 mmol/L磷酸盐缓冲液(p H 6.5)(35∶65)为流动相,检测波长270 nm,分离电压15 k V。米卡芬净在62.5~800μg/ml浓度范围内线性关系良好,平均回收率为99.6%,RSD为1.47%。     

棘白菌素的临床用药进展

棘菌素类抗真菌药物靶向作用于真菌细胞壁,具有独特的抗真菌效应。该类抗真菌药物如卡泊芬净（caspofungin）、米卡芬净（micafungin）和阿尼芬净（anidulafungin）对念珠菌属、曲霉菌属以及某些对唑类药物耐药的真菌菌种具有良好的抗菌活性。本文主要综述棘白菌素类药物临床应用进展。     

新型棘白菌素类抗真菌药阿尼芬净

阿尼芬净(anidulafungin,VER-002;LY303366),是第三代棘白菌素类的半合成抗真菌药,是两性霉素B的衍生物,由美国Vicuron制药公司研制,同其他棘白菌素类抗真菌药比较,分布容积更大,抗菌谱更广,目前在美国处于Ⅲ期临床试验阶段.介绍阿尼芬净的作用机制、药效学、药动学和临床评价.     

棘白菌素类抗真菌药临床应用研究进展

棘白菌素类（echinocandins）是21世纪初开发的最新型的一类广谱抗真菌药,其通过抑制真菌细胞壁的β-（1,3）-D-葡聚糖的合成,破坏真菌细胞壁的完整性,导致真菌细胞溶解死亡,是一种具有全新作用机理的抗真菌药物,对很多耐唑类药物的真菌仍具有良好的抗菌活性。因其通过干扰真菌细胞壁的合成而产生抗真菌作用,而人体细胞没有细胞壁,所以该类药对人体的毒性较低,是迄今为止安全性最高的一类抗真菌药物,具有抗菌谱广、抗真菌作用强、半衰期长、不良反应少、患者耐受性好等特点。该类药对念珠菌为杀菌剂,对曲霉菌属于抑菌剂,无交叉耐药性,是目前用于治疗全身性真菌感染的一线药物。目前国外已上市的棘白菌素类抗真菌药有三种：卡泊芬净、米卡芬净和阿尼芬净,其中卡泊芬净已在国内上市。本文对棘白菌素类药物的作用机制、适应证、抗菌谱、药动学、药物相互作用及不良反应等进行简要综述。     

棘白菌素B母核的分离纯化工艺研究

目的 研究大孔吸附树脂结合亲水性C18硅胶填料分离纯化棘白菌素B母核(ECBN)的工艺,有效去除色素及其他杂质,获得高纯度ECBN.方法 用HPLC检测方法,以ECBN的吸附量和洗脱率为指标,考察大孔吸附树脂和亲水性C18填料分离纯化ECBN的吸附性能和洗脱参数.结果 分离纯化ECBN的最佳吸附剂为大孔吸附树脂XAD-18和亲水性C18填料,洗脱剂分别为8％乙醇和3％甲醇.此工艺路线获得的成品纯度大于95％,总收率为65％.结论 该分离纯化方法为国内首次报道,工艺简单,使用有机溶剂少,分离效果好,适用于工业化生产.     

新型抗真菌药米卡芬净

对米卡芬净的作用机制、抗菌谱、药动学、药物相互作用、临床疗效有效性和安全性评价以及不良反应和耐受性的有关研究进行了综述.     

新型抗深部真菌感染药物——棘白霉素类

棘白霉素类(卡泊芬净、米卡芬净、阿尼芬净)抗真菌药是最新型的1种广谱抗真菌药,作用于真菌细胞壁,对于念珠菌属以及曲霉菌属均有效,使用安全性较高。该文主要综述了棘白霉素类药物的作用机制、适应证、抗菌谱、药动学、药物相互作用及不良反应。     

棘白菌素类抗真菌药物耐药机制研究

棘白菌素类新型抗真菌药物为β-(1,3)-D-葡聚糖合成酶抑制剂,在临床广泛应用.与唑类药物不同,霉菌和酵母菌对棘白菌素类药物的耐药涉及Fksl介导的耐药机制,本文简要综述棘白菌素类药物的真菌耐药机制研究进展.     

基于脱酰基酶转化阿尼芬净前体echinocandin B条件的研究

脱酰基酶可以将echinocandin B(ECB)脱去酰基侧链得到ECB母核,用于化学合成抗真菌药阿尼芬净。本研究考察了以二甲基亚砜(DMSO)助溶剂的转化体系脱酰基酶对ECB转化的各因素,建立了转化工艺(磷酸缓冲浓度：0.02mol/L,pH7.0,KCl浓度：0.68mol/L,DMSO浓度：15%,ECB浓度：450mg/L,加酶量：10%,转化温度：40℃)。为了提高转化效率,探索了新的转化体系即以β-环糊精助溶剂的转化体系,其优势为：当ECB与β-环糊精的浓度比为1:6时,ECB可在体系中完全溶解,底物浓度可以达到2g/L。     

Rapid separation and determination of structurally related anthraquinones in Rhubarb by pressurized capillary electrochromatography

A pressurized capillary electrochromatography (pCEC) with monolithic column has been developed for the rapid separation and determination of five structurally related anthraquinones in Rhubarb. The possibility of rapid separation resulted from the unique pore structure with high permeability and favorable mass transfer characteristics of the monolithic stationary phase. The effect factors such as organic modifier, acidity and concentration of running buffer, separation voltage were investigated to acquire the optimum condition. In the 220 nm wavelengths, the five anthraquinones could be baseline-separated rapidly within 5 min with the separation voltage of -20 kV in 10 mmol/L phosphate buffer (pH 6.2) containing 65% acetonitrile. The calibration graphs of rhein, aloe-emodin, emodin chrysophanol and physcion were linear by plotting the peak area against the analytes concentration over the range of 0.2-65, 0.1-30, 0.1-55, 0.5-30 and 0.5-55 microg/mL, respectively. The detection limits of five anthraquinones were ranged from 0.06 to 0.2 microg/mL and the recoveries of Rhubarb samples were about 81.3-86.4% (R.S.D.< or = 5.2%). This proposed method was successfully applied to determination of the five analytes in Rhubarb with satisfactory results.     

Separation and determination of coumarins in Fructus cnidii extracts by pressurized capillary electrochromatography using a packed column with a monolithic outlet frit

The pressurized capillary electrochromatography (pCEC) was utilized for the separation and determination of coumarins in Fructus cnidii extracts from 12 different regions. After a thorough study of analytical parameters such as acetonitrile content of the mobile phase, the concentration and pH of the buffer, and the applied voltage, a methodology was proposed to separate and determine six coumarins of F. cnidii extracts in less than 15 min. The experiments were performed in an in-house packed column with a monolithic outlet frit under the optimal conditions: pH 4.0 ammonium acetate buffer at 10 mM containing 50% acetonitrile at −6 kV applied voltage. The calibration curves were linear in the range of 10.0–100.0 μg/mL for bergapten, 20.0–200.0 μg/mL for imperatorin, 5.0–400.0 μg/mL for osthole, 10.0–100.0 μg/mL for 2′-acetylangelicin, 10.0–200.0 μg/mL for oroselone, and 10.0–200.0 μg/mL for O-acetylcolumbianetin. The correlation coefficients were between 0.9967 and 0.9995. With this pCEC system, fingerprints of F. cnidii extracts were preliminarily established to distinguish three types of coumarins by characteristic peaks, and the quality of various sources of raw materials was evaluated by determining the contents of six coumarins.     

应用加压毛细管电色谱技术检测鸡蛋中四环素类农兽药残留的研究

目的 建立了鸡蛋样品中四环素类农药多残留同时检测的加压毛细管电色谱分析方法。方法 以二氯甲烷为沉淀剂处理鸡蛋样品,运用加压毛细管电色谱法进行快速检测。以甲醇-乙腈-20 mmol/L草酸溶液(pH=4.25)(10：20：70,v/v/v)作流动相,等度洗脱,外加电压为-4kV,检测波长为270 nm。结果 经方法学考察,4种四环素类药品土霉素、4-差向金霉素、盐酸金霉素、强力霉素在各自标准曲线浓度范围内线性良好,R2均在0.993以上。回收率为80.46％～100.49％,日内和日间精密度的相对标准偏差(RSD)均低于5％。结论 该方法简单方便,重现性好,准确可靠,回收率较高,适用于鸡蛋样品中抗生素多残留的测定。     

毛细管区带电泳法分离盐酸美沙酮对映体

目的:采用毛细管区带电泳法对盐酸美沙酮对映体进行了拆分。方法:比较了3种衍生化β-环糊精为手性添加剂的分离效果,对缓冲液的pH及浓度、手性添加剂的浓度、柱温、分离电压等方面进行了考察及优化。结果:确定采用75μm×75 cm未凃渍石英玻璃管柱,运行缓冲液为含50 mmol.L-1的Tris和10 mmol.L-1的HP-β-环糊精的水溶液(以磷酸调节pH至2.0),分离电压为25 kV,柱温25℃,检测波长为205 nm,结论:该方法简便、快速,在15 min内分离度可达到1.9。     

棘白霉素B脱酰基酶高产菌株的选育及转化条件优化

目的 为提高棘白霉素B脱酰基酶产生菌的转化率。方法 采用常压室温等离子体诱变复合链霉素抗性筛选转化高产菌株;对新菌株转化棘白霉素B的底物溶剂、转化温度和转化时间等条件进行优化。结果 获得稳定突变株ZH-6-78比出发菌的转化率提高了26.7%;优化后转化条件为：转化温度35℃,转化时间24h。结论 高产菌种在优化的转化条件下,转化率达到90%,比出发菌原工艺提高了50.0%,为工业化生产奠定基础。     

手性功能化纳米材料涂层的毛细管/芯片电色谱手性拆分研究进展

目的阐述近年来基于手性功能化纳米材料涂层的毛细管/芯片电色谱手性拆分相关研究进展。方法归纳国内外最新的文献报道,对相关手性功能化纳米材料的理化性质及基于手性功能化纳米材料涂层的毛细管/芯片电色谱手性拆分研究进行综述。结果将手性功能化纳米材料涂层用于毛细管/芯片电色谱手性拆分中可以显著提高对映体的分离效果。结论手性功能化纳米材料涂层在毛细管/芯片电色谱手性分离领域具有良好的发展前景。     

Echinocandin B的分离纯化工艺研究

目的开发echinocandin B的分离纯化工艺。方法采用大孔吸附树脂吸附解吸-溶剂萃取-溶剂结晶的分离纯化方法。结果 HZ830树脂为最佳吸附剂,吸附流速选择为2BV/h,洗脱剂为85%乙醇,结晶溶剂为丙酮。在此实验条件下所得产品纯度≥95%,提取总收率大于60%。结论此方法简单,分离效果好,适于工业化生产。     

以盐酸去甲万古霉素为手性选择剂毛细管电泳法分离西替利嗪对映体

目的:建立以阳离子表面活性剂碘化四丁基铵为电渗流改性剂,以盐酸去甲万古霉素为手性选择剂的毛细管电泳法分离西替利嗪对映体。方法:考察了盐酸去甲万古霉素浓度、Tris 浓度和缓冲液 pH 对分离的影响,对分离条件进行了优化。结果:在含0.04 g·L~(-1)碘化四丁基铵和1.0 mmol·L~(-1)盐酸去甲万古霉素的25 mmol·L~(-1)Tris 磷酸缓冲液(pH 4.5)的运行电解质体系中,西替利嗪对映体在分离电压为20 kV 的条件下得到良好分离,西替利嗪对映体分离度达1.8。结论:本法可用于西替利嗪对映体的分离。     

Rapid analysis of trace levels of flavins by pressurized capillary electrochromatography-laser induced fluorescence detection with sulfonated N-octadecyl methacrylate monolith

In this paper, pressurized capillary electrochromatography (pCEC) with laser induced fluorescence detection (LIF) was demonstrated as a viable approach for the separation and determination of trace flavins in human plasma, where flavins tend to be degraded ex vivo. Using a sulfonated N-octadecyl methacrylate monolithic column in isocratic pCEC separation, symmetrical peak shapes and rapid separation could be obtained in a weakly acidic mobile phase. Baseline separation of riboflavin, flavin mononucleotide and flavin adenine dinucleotide could be achieved within 4.5 min in a mobile phase containing 60% (v/v) acetonitrile and 40% (v/v) of 20 mmol L⁻¹ phosphate buffer (pH 4.0), with −22 kV of applied voltage and 290 psi of supplementary pressure and 0.02 mL min⁻¹ of flow rate. Based on a 473 nm laser diode double pumped solid state source, flavins could be determined by LIF with the detection limit (LOD) as low as 0.5 nmol L⁻¹ (S/N = 3). The concentration ranges were 0.005–2 μmol L⁻¹ for RF and FMN, and 0.02–40 μmol L⁻¹ for FAD. Owing to the weakly acidic condition selected in this experiment, the high fluorescence quantum yields and good stability of flavins contributed to a preferable analysis. Combined with a simple clean-up procedure, this method has been proved to be effective for the rapid and selective analysis of trace levels of flavins in human plasma without sample preconcentration.