高效液相色谱-串联质谱法测定猪尿中秋水仙碱

提出了猪尿中秋水仙碱的液相色谱-串联质谱检测方法。采用pH8.0磷酸盐缓冲溶液碱化5mL尿样,用乙酸乙酯提取两次,合并提取液,于40℃吹氮蒸干,用2 mL流动相溶解残渣。在色谱分离中选用Sunfire C_(18)色谱柱,流动相为甲醇-2mmol·L~(-1)乙酸铵溶液(含0.1%甲酸)混合溶液(40+60),流量为0.2mL·min,进样量为10μL。质谱测定中采用电喷雾正离子模式离子化、多反应监测模式测定秋水仙碱,外标法定量。猪尿中秋水仙碱在1～50μg·L~(-1)内呈线性,测定下限(10S/N)为0.2μg·L~(-1),方法的回收率为90%～100%,相对标准偏差(n=6)小于9.0%。     

高效液相色谱法测定盐酸甲氯芬酯胶囊的含量

建立了用高效液相色谱测定盐酸甲氯芬酯胶囊含量的方法.采用Hypersil C18柱(5 μm,4.6 mm i.d.×200 mm),流动相为V(乙腈)∶V(0.12% NH4HCO3-0.50%三乙胺)溶液＝33∶67 (甲基磺酸调pH至3.0),流速: 1.0 mL/min,检测波长为225 nm.盐酸甲氯芬酯的线性范围为1.632～163.2 μg/mL,平均回收率为99.67%,RSD=1.8% (n=9).     

高效液相色谱法测定人血浆中奥氮平的浓度

 用高效液相色谱法测定了人血浆中奥氮平的浓度。色谱条件 :采用岛津LC 6A型高效液相色谱仪 ;色谱柱为ZorbaxODS (15 0mm× 4 6mmi d ,粒径 5 μm) ;流动相为V(5 0mmol/L磷酸钠缓冲液 ,pH 7 2 )∶V(甲醇 )∶V(乙腈 ) =12∶10∶3的溶液 ;检测波长为 2 70nm ;流速为 1 0mL/min ;柱温 40℃ ;灵敏度 0 0 0 5AUFS ;纸速 2mm/min。实验结果显示 ,在上述条件下 ,该方法的线性范围为 15 μg/L～ 12 0 0 μg/L(r =0 9988) ,最低检测限为 3μg/L ,血浆中奥氮平的平均回收率为 (97 0 2± 3 11) % ,测定结果的日内平均相对偏差为 3 86 % (n =15 ) 。     

超临界二氧化碳萃取秋水仙碱（英文）

利用超临界二氧化碳对秋水仙块根（经粉碎）中的秋水仙碱进行了萃取,采用高效液相色谱法对萃取出的秋水仙碱的质量分数进行了测定。实验选择40℃20～40MPa作为超临界苹取的操作条件,采用体积分数为95％的乙醇在索氏提取器中对样品进行了对比苹取实验。结果表明:不加浸泡剂进行浸泡处理的样品中的秋水仙碱很难被超临界二氧化碳萃取,在40℃,35MPa条件下,消耗1．28mol的二氧化碳只得到3％的萃取率。加入极少量的有机溶剂浸泡处理样品15min后再进行超临界萃取,可以极大程度地提高秋水仙碱的萃取率。在所选的浸泡剂乙醚、丙酮、乙腈、甲醇、乙醇中,乙醇浸泡效果最佳,经浸泡后,在40℃,35MPa条件下进行超临界二氧化碳萃取,仅消耗0．21mol的二氧化碳,一次苹取的秋水仙碱萃取率就达80％。秋水仙碱的质量分数从原植物样品中的0．18％提高到萃取物中的6．92％,提高了近38倍,而有机溶剂乙醇提取只能达到0．98％,仅提高了5倍。超临界苹取后的样品经简单的过滤即可进样进行HPLC分析,有机溶剂乙醇提取则需要氯仿苹取、溶剂回收等样品制备过程。HPLC分离测定秋水仙碱采用苯基柱,甲醇-水梯度洗脱,流速0．5mL／min,UV检测λ=353um,经选择,其最佳条件为：从V（甲醇）：V（水）=50：50开始,15min内到V     

超高效液相色谱-三重四极杆质谱法测定化妆品中秋水仙碱、秋水仙胺和秋水仙碱苷EI北大核心CSCD

建立了一种同时测定化妆品中秋水仙碱、秋水仙胺和秋水仙碱苷的超高效液相色谱-三重四极杆质谱分析方法。试样经正己烷饱和的甲醇-乙腈(1∶1,V/V)混合溶剂超声提取,蜡基类样品经正己烷溶解分散后提取,膏霜类样品在提取液中加入乙酸铵以改善样品乳化情况,提取液离心过滤后,以5 mmol/L乙酸铵(含0.1%甲酸)-甲醇作为流动相梯度洗脱,经ACQUITY UPLC BEH C_(18)色谱柱分离后,采用超高效液相色谱-三重四极杆质谱在电喷雾正离子电离模式和多反应监测(MRM)模式检测。3种化合物在0.5~10μg/L范围内线性关系良好,相关系数大于0.995,检出限为3.0μg/kg,定量限为10.0μg/kg,在10.0,20.0,100μg/kg 3个加标水平下的平均回收率为81.5%~109.2%,相对标准偏差为0.5%~8.7%。该方法适用于化妆品中秋水仙碱、秋水仙胺和秋水仙碱苷的测定。     

高效液相色谱法测定抗癌原料药芳香异羟肟酸二丁基锡

建立了反相高效液相色谱法测定有机金属抗癌原料药芳香异羟肟酸二丁基锡. 色谱条件: Diamonsil ODS 色谱柱 (4.6 mm i.d.×200 mm, 5 μm), 以V(甲醇)∶V(水)＝35∶65, pH 2.5为流动相, 流速为1.0 mL/min, 紫外检测波长232 nm, 柱温25 ℃. 吲达帕安为内标测定芳香异羟肟酸二丁基锡. 结果表明: 线性范围为4.08～102 μg/mL, 回归方程为f=0.02253ρ-0.005843, r=0.9998 (n=7), 方法精密度RSD为0.89% (n=6), 稳定性的RSD为0.56% (n=9), 重复性的RSD为0.90% (n=9), 平均回收率为100.5% (n=9, RSD=1.1%). 方法可用来测定芳香异羟肟酸二丁基锡.     

荧光法研究秋水仙碱和牛血清白蛋白的相互结合作用

用荧光光谱法、分光光度法研究了水溶液中秋水仙碱与牛血清白蛋白 (BSA)的相互结合反应。研究表明 :二者以摩尔比 1∶1牢固结合 ,其平衡常数K0 =1.92× 10 5L/mol。根据F rster非辐射能量转移机理 ,求算了给体 (BSA)与受体 (秋水仙碱 )间距离r =3.6 3nm和能量转移效率E =0 .17。实验表明 :秋水仙碱与牛血清白蛋白的相互结合作用为单一的荧光静态猝灭过程     

反相高效液相色谱法测定兔血清中拉莫三嗪的浓度及其动物药代动力学研究

 用反相高效液相色谱法测定了兔血清中抗癫痫药拉莫三嗪的浓度 ,以苯乙酮作内标 ,甲醇 磷酸二氢钾 ( 0 0 5mol/L ,pH 3 8)为流动相 ,经YWG ODS柱 ( 2 0 0mm× 3 6mmi d )分离 ,波长为 3 0 5nm的二极管阵列检测器检测 ,结果血药浓度在 0 2 5mg/L～ 5 0mg/L范围内线性良好 (r =0 9998) ,平均回收率为 93 78% ,标准误差为2 66% ,日内和日间RSD分别为 2 4 % (n =9)和 5 3 % (n =5 )。取家兔经胃给药 ( 2 5mg/kg) ,在设定时间抽血 ,测定动物药代动力学参数 ,经 3P87药代动力学程序包拟合 ,结果为Cmax=9 3 4 μg/mL ,t1/ 2 (Ke) =8 78h。     

高效液相色谱法测定痹祺胶囊中马钱子碱、士的宁和丹酚酸B

建立了同时测定痹祺胶囊中马钱子碱、士的宁和丹酚酸B含量的方法.采用反相高效液相色谱法,色谱柱:Inertsil ODS-3(250 mm×4.6 mm,5 μm);流动相:V(乙腈): V(10 mmol/L己烷磺酸钠的磷酸盐缓冲液,pH 2.86)=22: 78;流速: 1.0 mL/min;检测波长: 254 nm;柱温:25 ℃.马钱子碱、士的宁和丹酚酸B的线性范围分别为2.7～32 mg/L(r=0.9998)、2.6～31 mg/L(r=0.9999)和12.7～151 mg/L(r=0 9991);平均加样回收率(n=6)分别为99.3%、95.6%和98.9%;RSD分别为2.30%、3.68%和1.82%.本方法简便快捷,结果准确,可用于痹祺胶囊的质量控制.     

毛细管色谱法测定芳香异羟肟酸二丁基锡中有机溶剂残留量

建立了毛细管气相色谱法测定芳香异羟肟酸二丁基锡中的有机溶剂残留量.采用HP-5 (5%苯基甲基聚硅氧烷)毛细管色谱柱,FID检测器.载气为N2,柱温为45 ℃,检测器温度为160 ℃,流速为1.2 mL/min,分流比为20∶1,四氢呋喃为内标进行测定.结果表明: 甲醇、无水乙醇、三氯甲烷、正己烷、苯的线性范围分别为: 7.9～63.2 μg/mL (r=0.9991),8.69～69.52 μg/mL (r=0.9999),19.77～158.16 μg/mL (r=0.9990),295.6～2364.8 μg/mL (r=0.9996),8.74～69.92 μg/mL (r=0.9996),平均回收率分别为104.2%、 98.8%、 97.9%、 103.1%和102.0%,RSD分别为1.6%、 2.3%、 2.1%、 3.8%和1.9% (n=9);检出限分别为3.16、 3.47、 23.90、 1.97和0.87 ng.方法可用于有机溶剂残留量的测定.     

秋水仙碱分析方法的研究概述

针对秋水仙碱作为治疗急性痛风性关节炎药物的应用,综述了国内外秋水仙碱测量方法研究进展,重点介绍了伏安法、高效液相色谱法、高效液相色谱-质谱联用法等的研究情况.     

Effect of Deacetylation on Statmhokinetic Activity of Colchicine Derivatives

Screening of colchicine derivatives that was carried out with the tumor culture CaPa in a cytotoxicity test showed that deacetylation decreases the cytotoxicity and increases the mitotic index. Lower toxicity of the deacetylated colchicine derivatives was also noted.     

Polymorphism of Inclusion Complexes and Unsolvated Hosts. III. Dimorphism of the Alkaloid Colchicine Hydrate. The Structure of Colchicine Monohydrate

The alkaloid colchicine forms, in addition to the previously known dihydrate host–guest complex, a monohydrate complex. The crystal structure of the monohydrate was determined by direct methods and refined to a final R value of 0.046 for 1425 observed reflections. Crystal data are: orthorhombic, space group P2 12 12 1, a = 9.145(2) Å; b = 13.270(3) Å; c = 17.942(4) Å, V = 2177(1) Å3, Z= 4, Dx = 1.22 g cm-3, T = 293 K. The conformation of the molecule is practically identical with the conformation in the dihydrate inclusion complex. Water molecules show proton donor as well as proton acceptor properties and are hydrogen bonded with the three colchicine molecules giving rise to the three dimensional H-bonded network.     

An Optical-Fiber Sensor for Colchicine Using Photo-Polymerized N-Vinylcarbazole

An optical-fiber sensor for colchicine has been fabricated. It is based on the use of the fluorescent dye N-vinylcarbazole, which contains a vinyl unit making it photo-copolymerizable in the presence of a monomer, 1,2-cyclohexanediol diacrylate, on a quartz surface of an optode slide. Prior to the photo-polymerization process the quartz surface was modified with -(methacryloxy)propyl trimethoxysilane to introduce vinyl units. A dye-incorporating optode membrane tightly immobilized on the quartz surface was prepared in this way. The spectral characteristics of the optode for sensing colchicines were evaluated, and the response mechanism was discussed. The optode membrane has a relatively long lifetime and short response and recovering time. The linear range of determining colchicine is from 1.0×10^–6 to 4.0×10^–3molL^–1. The sensor was applied to determine the colchicine content in pharmaceutical tablets the results of which agreed with those obtained by the pharmacopoeia method.Received December 12, 2002; accepted March 25, 2003 Published online July 16, 2003     

取代薁酮的反应性研究: 秋水仙碱合成新策略的探索

本工作以探索秋水仙碱(Colchicine)合成新策略为目标, 研究了烷氧基取代薁酮2a的独特反应性, 发现了一系列罕见的重排过程和环庚三烯(Tropilidene)降蒈二烯(Norcaradiene)互变异构现象, 探讨了相应的反应机理.     

某些芳香族氨基酸作探针荧光猝灭法测定秋水仙碱

在适当的酸性介质中, 秋水仙碱(COL)能与色氨酸(Trp)、酪氨酸(Tyr)和苯丙氨酸(Phe)等芳香族氨基酸反应并形成结合产物, 此时将引起上述氨基酸的荧光发生猝灭, 最大猝灭波长分别位于350 nm (Trp), 304 nm (Tyr), 284 nm (Phe). 其荧光猝灭值(ΔF)在一定范围内与秋水仙碱成正比. 当用Trp和Tyr作探针时, 荧光猝灭法测定秋水仙碱具有高灵敏度, 其检出限分别为15.1 ng/mL (3.78×10^－8 mol/L)和19.8 ng/mL (4.96×10^－8 mol/L). 文中研究了适宜的反应条件和影响因素, 考察了共存物质的影响, 表明方法有良好的选择性, 可用于秋水仙碱的测定. 文中讨论了复合物的组成、结合力和结合模式. 通过温度的影响以及Stern-Volmer作图, 判断该反应为静态猝灭反应, 它们的结合常数(K)在25 ℃时分别为9.7×10⁶ (COL-Trp), 8.9×10⁶ (COL-Tyr)和6.3×10⁵ (COL-Phe). 其作用力主要是芳基堆集作用和氢键结合作用, 而芳基堆集作用是发生荧光猝灭的主要原因.     

秋水仙碱水解产物与某些酸性磺酞类染料相互作用的 共振瑞利散射光谱及其分析应用

在pH 2.9～4.6 Britton-Robinson (BR)缓冲溶液中, 秋水仙碱的水解产物(H-COL)能与溴酚蓝(BPB)、溴甲酚绿(BCG)、溴百里酚蓝(BTB)和百里酚蓝(TB)等酸性磺酞类染料(ASPD)反应形成1∶1的离子缔合物, 此时将引起共振瑞利散射(RRS)的急剧增强, 并产生新的RRS光谱. 秋水仙碱水解产物与溴酚蓝、溴甲酚绿、溴百里酚蓝和百里酚蓝形成离子缔合物的最大散射波长分别位于327, 311, 305和306 nm处. 散射增强(ΔI)与秋水仙碱浓度在一定范围内成正比, 不同体系对于秋水仙碱的检出限(3σ)分别为12.3, 15.1, 16.4和20.0 ng•mL－1 (TB). 研究了适宜的反应条件, 考察了共存物质的影响, 表明方法有较好的选择性. 基于秋水仙碱水解产物与酸性磺酞类染料离子缔合物的反应, 发展了一种较灵敏, 且简便、快捷测定秋水仙碱的新方法. 方法用于片剂、黄花、血清和尿样中秋水仙碱的测定, 获得了满意的结果.     

Electrochemical Determination of Natural Drug Colchicine in Pharmaceuticals and Human Serum Sample and its Interaction with DNA

Colchicine (COLC) is a natural toxic product and secondary metabolite most commonly used to treat gout. In this study, its electrochemical behavior and determination was investigated by employing modification‐free boron‐doped diamond electrode (BDDE). Besides, its interaction with DNA was monitored using electrochemical methods. It was found that oxidation of this compound proceeds in two steps, where first sharp and well defined oxidation peak occurs at potential of around 1.19 V, and second one at around 1.37 V, in Britton‐Robinson buffer solution at pH 7.5. Wide dynamic range from 1 to 100 μM was obtained with a detection limit (3σ_intercept/slope) of a 0.26 μM, based on the evaluation of first oxidation peak using differential pulse voltammetry. The proposed method was also found to be suitable for monitoring interaction of this drug with DNA as important segment for medical use. Concerning the validation, the analytical procedure shows excellent selectivity and sensitivity toward COLC detection and after method development it was successfully used for its quantification in pharmaceutical preparation and human serum sample, with satisfactory recovery. Obviously, this approach can be promising replacement for time‐consuming and expensive separation methods.     

A New Insight into Toxicity of Colchicine Analogues by Molecular Docking Analysis Based on Intestinal Tight Junction Protein ZO-1

Colchicine (COL) is a well-known plant alkaloid long used for medical purposes due to the selective anti-inflammatory effect on acute gouty arthritis. It is also a kind of mitosis toxin with strong inhibitory effects of cell division and is therefore being applied to the treatment of various cancers. However, this product shows a variety of adverse effects that are significantly correlated with the dosage and have attracted much attention. For the first time, the present work obtained a new insight into the gastrointestinal toxicity of colchicine analogues by molecular docking analysis, which was based on the 3D structure of intestinal tight junction protein ZO-1 and the ligand library containing dozens of small-molecule compounds with the basic skeleton of COL and its metabolites. The binding energy and mode of protein–ligand interaction were investigated to better understand the structure–toxicity relationships of COL analogues and the mechanism of action as well. Cluster analysis clearly demonstrated the strong correlation between the binding energy and toxicity of ligand molecules. The interaction mode further revealed that the hydrogen bonding (via the C-7 amide or C-9 carbonyl group) and hydrophobic effect (at ring A or C) were both responsible for ZO-1-related gastrointestinal toxicity of COL analogues, while metabolic transformation via phase I and/or phase II reaction would significantly attenuate the gastrointestinal toxicity of colchicine, indicating an effective detoxication pathway through metabolism.     

Synthesis of Lipid Derivatives of Colchicine

The synthesis of glycero-lipids linked to colchicine derivatives is reported. The lipid structures are designed to perform two-dimensional crystallization experiments with tubulin, the structural subunit protein of microtubules.