Novel method for the determination of five carbamate pesticides in water samples by dispersive liquid-liquid microextraction combined with high performance liquid chromatography

A novel method for the determination of five carbamate pesticides （metolcarb, carbofuran, carbaryl, isoprocard and diethofencard） in water samples was developed by dispersive liquid-liquid microextraction （DLLME） coupled with high performance liquid chromatography-diode array detector （HPLC-DAD）. Some experimental parameters that influence the extraction efficiency were studied and optimized to obtain the best extraction results. Under the optimum conditions for the method, the calibration curve was linear in the concentration range from 5 to 1000 ng mL^-1 for all the five carbamate pesticides, with the correlation coefficients （r^2） varying from 0.9984 to 0.9994. Good enrichment factors were achieved ranging from 80 to 177- fold, depending on the compound. The limits of detection （LODs） （S/N = 3） were ranged from 0.1 to 0.5 ng mL^-1. The method has been successfully applied to the analysis of the pesticide residues in environmental water samples.

Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Optimization strategies for separation of sulfadiazines using Box-Behnken design by liquid chromatography and capillary electrophoresis

Development of effective chromatographic or electrophoretic separation involves judicious deciding of selection of optimal experimental conditions that can provide an adequate resolution at a reasonable run time for the separation of interested components. Box-Behnken factorial design was effectively applied for the separation optimization of eight structurally related sulfonamides using capillary zone electrophorosis and reverse high performance liquid chromatography. Optimum values for volume ratio of THF to H2O in eluent, column temperature and flow rate of eluent are found as 12 to 88, 35℃ and 1.0 mL/min, respectively. Box-Behnken modified optimization model is extended to separation by capillary electrophoresis (CE). While using CE, a satisfactory separation is achieved with a minimum resolution larger than 1.0 for a separation time less than 10 min.

益肾固精暖脐贴制备工艺的研究

目的研究益肾固精暖脐贴的提取工艺及其成型工艺。方法以蛇床子素、羟基-α-山椒素、金丝桃苷以及浸膏得率为考察指标,以提取时间、提取溶媒倍数、乙醇浓度为考察因素,运用Box-Behnken设计-响应面法优选出最佳提取工艺;以凝胶贴膏的初黏力、持黏力、剥离强度、感官评价为指标,采用D-最优混料设计优选益肾固精暖脐贴的最佳成型工艺。结果益肾固精暖脐贴的最佳提取工艺为回流提取120 min,提取溶媒倍数为12,乙醇浓度为57%,同法提取2次;最佳成型工艺的质量比为NP700占6.00%、PVP-k90占0.60%、甘羟铝占0.15%、填充剂5.76%、EDTA-2Na占0.10%、甘油占21.76%、柠檬酸占0.20%、水和药液总量占65.43%。结论该实验优选的益肾固精暖脐贴的提取工艺及其凝胶贴膏的制备工艺稳定可行,可为该产品的进一步开发利用提供参考。

基于分析质量源于设计理念的鱼腥草芩蓝合剂水提浓缩液中7种化学成分的定量研究

该研究基于分析质量源于设计理念,建立了测定鱼腥草芩蓝合剂水提浓缩液中绿原酸、隐绿原酸、槲皮苷、黄芩苷、汉黄芩苷、汉黄芩素、黄芩素含量的高效液相色谱法。使用鱼骨图识别可能影响方法属性的方法参数,通过确定性筛选设计明确关键方法属性为槲皮苷与前后峰的分离度和最后一个色谱峰的保留时间。首先采用加权决定系数法筛选出关键方法参数为第一梯度结束时间、第三梯度结束时间、第四梯度结束时间、柱温、流速,随后通过逐步回归法建立关键方法参数与关键方法属性之间的定量模型,模型的决定系数(R2)值超过0.90。采用不达标概率值定量表征不同参数组合下的风险大小,根据风险大小确定方法可操作设计区域,并成功验证了其可靠性。确定的色谱条件为:Sunshell-C_(18)(150 mm×4.6 mm,2.6μm)色谱柱,流动相为0.60%甲酸水(A)-乙腈(B),梯度洗脱,检测波长为260 nm和326 nm,流速0.80 mL/min,柱温39.0℃,进样量10μL。对优化后的分析方法进行方法学验证,方法线性良好,精密度、稳定性和重复性试验结果良好,相对标准偏差均在5.0%以内,7种成分的平均回收率为92.6%~106%,相对标准偏差在2.0%内。

基于分析质量源于设计的银杏叶提取液中有机酸含量分析方法

基于分析质量源于设计(AQbD)理念建立了银杏叶提取液中6种有机酸含量的高效液相色谱-二极管阵列检测器(HPLC-DAD)分析方法。首先,使用鱼骨图确定潜在的关键方法参数,然后通过确定性筛选设计辨识关键方法评价指标,再采用逐步回归法建立关键方法参数与关键方法评价指标之间的定量关系,采用达标概率法建立了方法的可操作设计区域并成功验证。用Plackett-Burman设计评价分析方法的耐用性。该方法通过方法学验证,可用于银杏叶提取液中有机酸含量的测定。

牛膝盐炙工艺研究

目的:探究牛膝最佳盐炙工艺。方法:采用超高效液相色谱串联质谱技术,以牛膝补肾壮骨的5种有效成分——β-蜕皮甾酮、25R-牛膝甾酮、25S-牛膝甾酮、人参皂苷R0及竹节参皂苷Ⅳa为评价指标,采用单因素和Box-Behnken设计方法,选择加盐量、炒制温度和炒制时间为考察因素,优化牛膝的盐炙工艺。结果:牛膝盐炙最佳工艺为2%加盐量,炒制温度100℃,炒制时间20 min。结论:所得牛膝盐炙工艺稳定可靠,有利于提高牛膝盐炙品的质量均一性。

活血消痛酒质量标准及制备工艺研究

目的:建立活血消痛酒质量标准并对其制备工艺进行优化。方法:采用薄层色谱法(TLC)对活血消痛酒的君药两面针、三七和臣药血竭鉴别;采用高效液相色谱法(HPLC)测定三七苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb_(1)的含量。色谱柱为Waters X Bridge Shield RPC18(4.6×250 mm,5μm),流动相:水(A)-乙腈(B),梯度洗脱(0~15 min,20%~23%B;15~25 min,23%B;25~35 min,23%~40%B;35~36 min,40%~90%B);流速为1 mL/min;柱温30℃;检测波长为203 nm;进样量为10μL;采用中心组合设计(CCD)响应面法对活血消痛酒工艺优化。结果:两面针、三七、血竭薄层色谱特征斑点清晰,分离度良好,阴性对照无干扰;三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb_(1)线性范围内良好(r^(2)>0.999),阴性对照无干扰、精密度、稳定性、重复性、加样回收率结果良好,10批活血消痛酒的三种皂苷含量范围为0.4225~0.5982 mg/mL;采用CCD法结合实际情况得出最优工艺:含醇量为42%、物料比为16.7 mg/mL、浸泡时间为7d。结论:该研究建立的方法操作简便、准确可靠,可用于活血消痛酒的质量标准研究;本研究通过中心组合设计法,科学、全面准确地找到了活血消痛酒的最优工艺。

微柱离心-HPLC法测定枸橼酸他莫昔芬脂质体包封率的方法研究

建立了枸橼酸他莫昔芬脂质体(TAM-LIPs)包封率的测定方法。采用薄膜分散法制备TAM-LIPs,用葡聚糖凝胶G-50填充微柱,以水为洗脱液,离心分离TAM-LIPs与游离药物后,通过高效液相色谱法(HPLC)测定枸橼酸他莫昔芬浓度并计算包封率,利用Box-Behnken效应面法优化微柱离心法测定条件。优化条件下,HPLC法能准确测定药物含量,洗脱曲线表明洗脱4次可以使载药脂质体完全被洗脱,可实现脂质体与游离药物良好分离,验证制备的3批TAM-LIPs包封率的RSD值满足方法学要求。建立的葡聚糖凝胶微柱离心-HPLC法测定结果准确,操作简单快捷,可用于TAM-LIPs包封率的准确测定。

Study on chromatographic fingerprint of sarcandra glabra (Thunb.) by microwave-assisted extraction coupled to HPLC/DAD

Microwave-assisted extraction(MAE)was used for extraction of effective components of sarcandra glabra(Thunb.),and then chromatographic fingerprint of sarcandra glabra(Thunb.)was studied by high performance liquid chromatography/diode array detector(HPLC/DAD).The conditions of MAE were optimized by an orthogonal experiment,and then the authentication and validation of the chromatographic fingerprint were conducted.Nine peaks were identified as common peaks in the fingerprint chromatograms,and isofraxidin was considered as a reference compound and quantified.Relative standard deviations of retention time and peak area of each component were less than 3% and 8%,respectively.Similarity and difference analysis were conducted by use of PCA and relation coefficient.Twenty batches of sarcandra glabra(Thunb.)samples from two different producing areas could be classified into two different groups in the PCA model.The results showed that MAE-HPLC/DAD method was simple,efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra(Thunb.),which could provide more reliable and precise information for quality evaluation.

Extraction optimization of coumarins from radix angelicae pubescentis by HPLC-DAD coupled with uniform design

Uniform design was used to optimize extraction condition of direct refluence extraction of coumarins from the Chinese traditional medicine of radix angelicae pubescentis(Duhuo); the sum peak area of coumarins separated with high performance liquid chromatography(HPLC) at detection wavelength of 320nm was considered as detection index, two factors of solvent concentration and extraction time were mainly studied at extraction temperature of 85℃ and a volume ratio of solvent to sample of 10∶1. Optimal subclass, quadric polynomial step by step aggression and neural network method were applied to process the experimental results. The results show that the first and second methods give the same factors combination (concentration of ethanol: 95%, extraction time: 3.6 h) and the second method is much better than the first one. The extraction model is consequently developed.

高效液相在仪器分析设计性实验中的教学设计与实践

设计性实验是综合多门学科知识和实验原理来设计实验方案,要求学生能充分利用所学知识,去发现问题、解决问题。高效液相色谱仪为药学专业学生常用分析测试仪器,通过设计性实验方案,训练学生文献检索、实验方案设计、仪器使用、数据分析及论文撰写等多方面能力。实践结果表明,基于设计性的高校液相实验教学能够启发学生的积极性,激发学生的学习热情,有效提高实验教学质量。

超高效液相色谱-串联质谱法测定毛发中7种新型苯二氮[艹卓]类策划药物的含量

由于新型苯二氮[艹卓]类策划药物具有用量小、药效强、体内代谢快和含量低等特点,很难在受害人的血液和尿液中检出,而毛发具有检出时效长、易获取和易保存的优势,因此提出了题示方法.取剪至1~2 mm的毛发20 mg,加入1 m L含内标(1μg.L^(-1)阿普唑仑Gd5)提取剂,于-50℃预冷5min,冷冻研磨两次,每次90s,中间间隔60s,并离心5min,取上清液浓缩至干,用100μL甲醇复溶,经0.22μm有机滤膜过滤.滤液在Acquity UPLCHSST3色谱柱上进行分离,经不同体积比的0.1%(体积分数,下同)甲酸溶液G含0.1%甲酸的乙腈溶液的混合液为流动相进行梯度洗脱,采用电喷雾离子源正离子模式(ESI+)扫描,多反应监测(MRM)模式检测,以保留时间和特征离子定性,内标法定量.结果表明,7种新型苯二氮[艹卓]类策划药物(瑞马唑仑、科纳唑仑、氟阿普唑仑、氟溴唑仑、玻玛唑仑、依替唑仑、溴替唑仑)工作曲线的线性范围为1~500pg.mg^(-1),检出限(3S/N)为0.11~0.35pg.mg^(-1).按照标准加入法进行回收试验,回收率为90.2%~104%,日内精密度和日间精密度试验所得相对峰面积的相对标准偏差(n=6)分别为1.7%~4.7%和2.9%~7.0%.

基于正交试验设计的大黄甘草洗液提取工艺研究

目的对大黄甘草洗液的提取工艺进行优化,为中试放大生产提供质量控制依据。方法利用高效液相色谱法测定蒙花苷的含量,采用正交试验考察加水倍量、浸泡时间、提取时间及提取次数等因素对提取效果的影响。结果大黄甘草洗液的最佳提取工艺为药材加14倍量水浸泡1小时,提取2次,每次2小时。结论提取工艺优化后稳定可靠,可为该药的量产提供保证。

葛渣总黄酮提取及葛根素含量测定

目的:优化葛渣中总黄酮提取工艺条件,测定葛渣中葛根素含量。方法:以葛渣为原料、总黄酮得率为评价指标,采用L9(34)正交试验设计,考察乙醇体积分数、提取时间、固料比、提取温度对葛渣总黄酮提取效果的影响,同时采用高效液相色谱法(HPLC法)测定葛根素的含量。结果:葛渣中总黄酮的最佳提取工艺条件为以体积分数70%乙醇溶液为溶剂、提取150min、料液比1:40(g/mL)、提取温度80℃,此条件下的总黄酮得率最高,为32.41mg/g。在该提取条件下,葛根、葛渣和葛粉中总黄酮的含量分别为46.11、32.41mg/g和11.23mg/g;葛根、葛渣和葛粉中葛根素的含量分别为40.63、21.24mg/g和4.71mg/g。结论:葛渣中含有丰富的总黄酮和葛根素,该方法可作为从葛渣中提取总黄酮和葛根素的方法。

微波辅助萃取人参总皂苷与单体皂苷含量分析

通过单因素试验和正交试验对微波辅助萃取人参皂苷的工艺条件进行优化,采用分光光度法和高效液相色谱法对萃取物中的人参总皂苷及8种单体皂苷进行测定,考察不同提取条件下所得人参皂苷产率和组成的差异。结果表明:1)以总皂苷为提取目标时,最佳提取条件为萃取功率600 W、萃取温度45℃、萃取时间5 min、料液比1∶20,萃取3次;2)以8种单体皂苷为提取目标时,最佳提取条件为萃取功率300 W、萃取温度35℃、萃取时间15 min、料液比1∶25,萃取3次(提取率为0.98%);3)在最佳提取条件下,以体积分数80%甲醇为提取溶剂时,总皂苷提取率为6.02%,而单体皂苷提取率之和为0.43%;以水饱和正丁醇为提取溶剂时8种单体皂苷提取率达到0.71%。综上所述,人参质量评价和工艺优化结果与提取方法、提取条件、评价指标密切相关。

超临界CO_2萃取南美白对虾虾青素的工艺优化

为提高虾青素萃取物得率和虾青素纯度,采用超临界CO2萃取南美白对虾虾头废弃物中的虾青素,皂化后用高效液相色谱对虾青素含量进行测定。单因素试验确定萃取参数对虾青素萃取物得率和虾青素纯度的影响,然后进一步运用响应面法对萃取工艺参数进行优化。结果表明:超临界CO2萃取工艺参数对虾青素萃取物得率影响不显著(P>0.05),但对虾青素纯度影响显著(P<0.05);物料粒径40目,质量含水率9%时,响应面理论优化最佳萃取参数为萃取压力403.95Pa,萃取温度39.95℃,CO2流量1.16L/min,虾青素纯度为796.3g/g。结合实际应用,验证试验在萃取压力400Pa,萃取温度40℃,流量1.2L/min时,虾青素纯度为789.61g/g。研究结果可为虾青素的提取和纯化提供参考。

均匀设计法优选穿山龙的水解工艺条件

目的:优化穿山龙水解条件,为工业上提取薯蓣皂甙元提供工艺参数。方法:以薯蓣皂甙元的得率为指标,采用均匀设计法和高效液相色谱法探讨穿山龙水解的最佳工艺条件。结果:盐酸浓度为2.5 mol/L,料液比为1:5,反应温度为140℃,反应时间为4.0h,搅拌速率为120rpm,穿山龙的水解效果较好。结论:采用优化后的水解条件明显提高薯蓣皂甙元的得率,盐酸浓度对结果影响极大,应加以控制。

响应曲面法用于高效液相色谱优化分离磺胺的研究

通过响应曲面法对高效液相色谱分离磺胺的优化过程进行了研究,分别考察了分离的3个主要因素(柱温、流动相配比、流速)对最小分离度与分离时间的影响,建立了3个因素与最小分离度和最大保留时间的目标函数,从响应曲面图可优化预测因素水平范围内的最佳分离条件。在选定优化条件下:柱温为20℃,乙腈的体积分数19%,流速1.5 mL/min,8种磺胺组分可在8 min内达到基线分离。

应用质量源于设计的理念优化大鼠血浆中大黄蒽醌的分析方法

应用质量源于设计理念建立一种高效液相色谱-荧光检测法(HPLC-FLD)用于测定大鼠血浆中5种大黄蒽醌。用Plackett-Burman设计考察流动相中甲醇含量、pH值、流速、柱温和进样体积对色谱峰的分离度、理论塔板数、最末洗脱峰的保留时间和拖尾因子的影响,结果显示流动相中甲醇含量、流速和柱温对色谱系统的影响显著(p<0.05)。继而采用Box-Behnken设计结合响应面法考察上述三因素对分离度、保留时间和理论塔板数的影响。用Derringer渴求函数评价了响应值的综合作用。得出最优色谱条件为:以甲醇-0.1%(v/v)磷酸水溶液(81.4∶18.6,v/v)为流动相等度洗脱,流速1.1 mL/min,柱温31℃,荧光检测激发波长为440 nm,发射波长为540 nm。建立的模型显示良好的预测性。结果表明:质量源于设计的理念可有效地应用于优化高效液相色谱分析方法。