高效液相色谱-电喷雾-串联质谱研究四溴双酚A双(2-羟乙基醚)诱导大鼠嗜铬细胞瘤细胞呼吸链氧化磷酸化和能量代谢紊乱

四溴双酚A衍生物的毒理学研究亟须开展。已有研究发现四溴双酚A双(2-羟乙基醚)(tetrabromobisphenol A bis(2-hydroxyethyl ether),TBBPA-BHEE)可诱导大鼠嗜铬细胞瘤细胞(PC12)活性氧(reactive oxygen species,ROS)的生成。然而TBBPA-BHEE对PC12细胞线粒体呼吸链氧化磷酸化过程的干扰机制尚不明确,TBBPA-BHEE是否通过破坏线粒体功能干扰细胞能量代谢亟待进一步探讨。建立了基于HPLC-ESI-MS/MS分析PC12细胞内ATP、ADP、AMP及cAMP(cyclic AMP)浓度的方法,在此基础上评价了TBBPA-BHEE暴露对PC12线粒体呼吸链氧化磷酸化过程及能量代谢的影响。研究发现,TBBPA-BHEE可加速PC12线粒体呼吸链氧化磷酸化过程;TBBPA-BHEE诱导的PC12线粒体功能紊乱可引起细胞能量代谢紊乱。一方面揭示了TBBPA-BHEE对PC12潜在的毒性作用机制,另一方面也证实HPLC-ESI-MS/MS是研究细胞线粒体呼吸链氧化磷酸化及能量代谢过程的有力工具。     

柱后补偿-液相色谱-电喷雾式检测器测定饮料中的三氯蔗糖

A method was developed for the determination of sucralose in soft drink by liquid chromatography ( LC) tandem a charged aerosol detector( CAD) with post column compensation. The results showed that the...     

Determination of lipid content and stability in lipid nanoparticles using ultra high-performance liquid chromatography in combination with a Corona Charged Aerosol Detector

For many years, lipid nanoparticles (LNPs) have been used as delivery vehicles for various payloads (especially various oligonucleotides and mRNA), finding numerous applications in drug and vaccine development. LNP stability and bilayer fluidity are determined by the identities and the amounts of the various lipids employed in the formulation and LNP efficacy is determined in large part by the lipid composition which usually contains a cationic lipid, a PEG-lipid conjugate, cholesterol, and a zwitterionic helper phospholipid. Analytical methods developed for LNP characterization must be able to determine not only the identity and content of each individual lipid component (i.e., the parent lipids), but also the associated impurities and degradants. In this work, we describe an efficient and sensitive reversed-phase chromatographic method with charged aerosol detection (CAD) suitable for this purpose. Sample preparation diluent and mobile phase pH conditions are critical and have been optimized for the lipids of interest. This method was validated for its linearity, accuracy, precision, and specificity for lipid analysis to support process and formulation development for new drugs and vaccines.     

Investigation of polar organic solvents compatible with Corona Charged Aerosol Detection and their use for the determination of sugars by hydrophilic interaction liquid chromatography

A range of organic solvents (ethanol, isopropanol and acetone) has been investigated as alternatives to acetonitrile and methanol when used in conjunction with Corona Charged Aerosol Detection (Corona CAD). These solvents have been evaluated with regard to their effect on the response of the Corona CAD. Three dimensional response surfaces were constructed using raw data showing the relationship between detector response, analyte concentration and percentage of organic solvent in the mobile phase, using sucralose or quinine as the test analyte. The detector response was non-linear in terms of analyte concentration for all solvents tested. However, detector response varied in an approximately linear manner with percentage of organic solvent over the range 0–40% for ethanol or isopropanol and 0–80% for acetone and methanol. The chromatographic performance of the various solvents when used as aqueous–organic mobile phases was evaluated for isocratic and gradient separations of sugars and sugar alcohols by hydrophilic interaction liquid chromatography (HILIC) using an Asahipak NH2P-504E column coupled with Corona CAD detection. It was found that whilst acetonitrile provided the highest column efficiencies and lowest detection limits of the solvents studied, acetone also performed well and could be used to resolve the same number of analytes as was possible with acetonitrile. Typical efficiencies and detection limits of 5330 plates m⁻¹ and 1.25 μg mL⁻¹, respectively, were achieved when acetone was used as the organic modifier. Acetone was utilised successfully as an organic modifier in the HILIC separation of carbohydrates in a beer sample and also for a partially digested dextran sample.     

在线固相萃取法结合电雾式检测器测定黄芪及其复方中黄芪甲苷的含量

采用在线固相萃取高效液相色谱法,结合电雾式检测器,建立了快速测定黄芪及其复方中黄芪甲苷的方法。采用Acclaim Polar AdvantageⅡC18(50 mm×4.6 mm,3μm)为在线净化柱,以双三元液相色谱的右泵作为上样净化泵,甲醇-水为流动相,梯度洗脱,进行富集和净化;采用Acclaim C18(150 mm×4.6 mm,5μm)为分析柱,以双三元液相色谱的左泵作为分析泵,乙腈-水作为流动相,等度洗脱,流速为1.0 mL/min,采用中心切割的方式将目标物从在线净化柱转移至分析柱中。电雾式检测器( CAD)的雾化器温度为30℃;氮气压力为241.3 kPa。黄芪甲苷在药材及复方基质中获得较好分离,黄芪甲苷在4.0~80 mg/L范围内,相关系数r=0.9998,平均回收率为97.6％。实验表明,本方法快速、简便,专属性、重现性较好,测定结果可靠,可应用于黄芪及其复方中黄芪甲苷含量的检测。     

局部麻醉药的反相色谱安培法检测

本文系统研究了普鲁卡因,利多卡因,丁卡因等3种局部麻醉药的液相色谱安培法检测行法,并与液相色谱光度法检测结果进行了比较。     

银黄口服液的质量控制及其高效液相色谱指纹图谱的研究

利用高效液相色谱方法,建立了复方银黄口服液及其原料药材黄芩和金银花的指纹图谱,测定了银黄口服液中黄芩苷 和绿原酸的含量。以Lichrospher C18色谱柱(250 mm×4.6 mm i.d.,5 μm)为分离柱,以甲醇和0.1%磷酸水溶液为流动 相进行二元梯度洗脱,流速0. 6 mL/min,在254 nm波长下检测。采用夹角余弦和相关系数的方法,计算6批银黄口服液指 纹图谱的相似度均在0.99以上,表明银黄口服液的质量未见显著差异。将银黄口服液指纹图谱中的共有峰与原料药材黄芩 和金银花的指纹图谱中的共有峰相比较,对其归属进行确认,大部分共有峰可以匹配。     

液相色谱电化学检测抗结核药物—异烟肼

本文讨论了以无有机溶剂存在的水溶液为流动相异烟肼的液相色谱一薄层安培检测法。在所选定的实验条件下,检测限达2ng。应用本法分析了片剂中异烟肼的含量     

Universal response model for a corona charged aerosol detector

The universality of the response of the Corona Charged Aerosol Detector (CoronaCAD) has been investigated under flow-injection and gradient HPLC elution conditions. A three-dimensional model was developed which relates the CoronaCAD response to analyte concentration and the mobile phase composition used. The model was developed using the response of four probe analytes which displayed non-volatile behavior in the CoronaCAD and were soluble over a broad range of mobile phase compositions. The analyte concentrations ranged from 1μg/mL to 1mg/mL, and injection volumes corresponded to on-column amounts of 25ng to 25μg. Mobile phases used in the model were composed of 0-80% acetonitrile, mixed with complementary proportions of aqueous formic acid (0.1%, pH 2.6). An analyte set of 23 compounds possessing a wide range of physicochemical properties was selected for the purpose of evaluating the model. The predicted response was compared to the actual analyte response displayed by the detector and the efficacy of the model under flow-injection and gradient HPLC elution conditions was determined. The average error of the four analytes used to develop the model was 9.2% (n=176), while the errors under flow-injection and gradient HPLC elution conditions for the evaluation set of analytes were found to be 12.5% and 12.8%, respectively. Some analytes were excluded from the evaluation set due to considerations of volatility (boiling point <400°C), charge and excessive retention on the column leading to elution outside the eluent range covered by the model. The two-part response model can be used to describe the relationship between response and analyte concentration and also to offer a correction for the non-linear detector response obtained with gradient HPLC for analytes which conform to the model, to provide insight into the factors affecting the CoronaCAD response for different analytes, and also as a means for accurately determining the concentration of unknown compounds when individual standards are not available for calibration.     

Disposable Carbon Electrodes for Liquid Chromatographic Detection of Catecholamines in Blood Plasma Samples

This paper describes a comparative evaluation of disposable carbon electrodes and conventional glassy carbon electrodes. The detection of catecholamines was demonstrated in processed blood plasma and in the presence of catecholamine metabolites. Calibration plots of norepinephrine, epinephrine, 3,4‐dihydroxybenzylamine and dopamine were linear over three to four orders of magnitude with detection limits of 0.50, 0.73, 1.06, and 1.13 pg, respectively. The relative standard deviation of peak areas was ±2.3% for norepinephrine and ±5.3% for epinephrine from 30 injections of a 10 ng/mL mixed standard. Spike recoveries for norepinephrine, epinephrine, and dopamine from human blood plasma were 86±6%, 81±5%, and 77±4%, respectively.     

Rapid and Sensitive Method for Citrinin Determination Using High-Performance Liquid Chromatography with Fluorescence Detection

Citrinin is a toxic product of secondary metabolism of fungi, such as certain Aspergillus, Penicillium, and Monascus species that are usually contaminating cereals. A new sensitive liquid chromatographic method with fluorescence detection was developed, validated, and applied for citrinin determination. The method is based on reversed-phase separation at pH 2.5, where citrinin exhibits the highest fluorescence quantum yield. In this setup, no derivatization step is needed. The method shows linearity in the range between 0.2 μg/mL and 0.1 mg/mL. The detection limit reached is 90 ng/mL (3.6 × 10^?7 M). Validated method was successfully applied on analysis of spiked and real cereal samples.