Creation of reversed phase high-performance liquid chromatographic technique to assay platelet-activating factor

Objective: To establish a new assay for platelet-activating factor (PAF), to compare it with bio-assay; and to discuss its significance in some elderly people diseases such as cerebral infarction and coronary heart disease. Methods: To measure PAF levels in 100 controls, 23 elderly patients with cerebral infarction and 65 cases with coronary heart disease by reversed phase high-performance liquid chromatographic technique (rHPLC). Results:rHPLC is more convenient, sensitive,specific, and less confusing, compared with bio-assay. The level of plasma PAF in patients with cerebral infarction was higher than that in the controls (P<0.01), and in patients with coronary heart disease. Conclusion: Detection of PAF with rHPLC is more reliable and more accurate. The new assay has important significance in PAF research.

High Performance Liquid Chromatographic Determination of Phenolic Acids in Fruits and Vegetables

A simple isocratic HPLC technique has been developed for the quantitative analysis of phenolic acids (PAs) in fruits and vegetables. Nine benzoic and cinnamic acid derivatives were separated in less than 30 min, and the resolution was all more than 1.23. The ranges of linearity for PAs standards were 0.2-100 ng, even up to 600 ng (r = 0.983-1.000) and the detection limits were 0.02-0.24 mg/kg. Samples of fresh vegetables and fruits were extracted with 80% mcthanol and ethyl acetate, then purified with C18 Sep-Pak cartridge and determined by HPLC. This method was applied to the determination of PAs in 7 kinds of fruits and vegetables, i.e., apple, pear, Chinese cabbage, cauliflower, turnip, soybean sprout and white grape wine. The content of the 9 PAs varied widely in the 7 kinds ol'foods studied. The average concentrations ofchlorogenic acid in apple (100.2 mg/kg) and pear (30.8 mg/kg) were quite high, and sinapinic acid was remarkable (42.5 mg/kg) in Chinese cabbage, and protocatechuic acid had the highest concentration of all the PAs in white wine.

Determination of Trace Amount of Polycyclic Aromatic Hydrocarbons in Urban Sewage by Solid-phase Extraction Coupled with High Performance Liquid Chromatograph

[Method] This study aimed to determine trace amount of polycyclic aromatic hydrocarbons(PAHs) in urban sewage by using solid-phase extraction(SPE) coupled with high performance liquid chromatograph(HPLC).[Method] From the aspects of solid-phase extraction column,elution solvent,elution volume,elution speed and so forth,the test conditions of SPE-HPLC method were optimized,and trace amount of PAHs in urban sewage was determined.[Result] The optimized solid-phase extraction conditions were SUPELCLEAN LC-18 solid-phase extraction column,methylene dichloride as elution solvent,15 ml elution volume,2 ml/min elution speed,5 ml/min loading speed,1 000 ml water with 200 ml methanol loading volume.Under the optimized extraction conditions,the recovery was high,namely 76.3%-105.2%;relative standard deviation was 3.8%-6.0%,showing good precision;detection limit was low,only 0.000 8-0.048 0 μg/L.[Conclusion] This method is user-friendly,with high sensitivity and good precision,and suitable for continuous determination of a large volume of water samples.

Determination of Organic Acids in Root Exudates by High Performance Liquid Chromatography:Ⅱ.Influence of Several Testing Conditions

Effects of column temperature and flow rate on separation of organic acids were studied by determining nine low-molecular-weight organic acids on reversed- phase C18 column, using high performance liquid chromatography (HPLC) with a wavelength of UV (ultraviolet) 214 urn and a mobile phase of 18 mmol L-1 KH2PO4 buffer solution (pH 2.1). The thermal stability of organic acids was determined by comparing the recoveries of organic acids in different temperature treatments. The relationships between column temperature, flow rate or solvent pH and retention time were analyzed. At low solvent pH, separation efficiency of organic acids was increased by raising the flow rate of the solvent because of lowering the retention time of organic acids. High column temperature was unfavorable for the separation of organic acids. The separating effect can be enhanced through reducing column temperature in organic acid determination due to increasing retention time. High thermal stability of organic acids with low concentrations was observed at temperature of 40 ℃-45℃. Sensitivity and separation effect of organic acid determination by HPLC were clearly improved by a combination of raising flow rate and lowering column temperature at low solvent pH.

Preparative Optimization of Cellulose Microspheres Applied as Supports for High-Performance Liquid Chromatography

The aim of this work is optimizing the techniques to prepare pure cellulose microspheres, which are used as packing adsorbents for high-performance liquid chromatography. Thereupon, cellulose was dissolved in a pre-cooled NaOH/urea solution, from which various-size microspheres were prepared. The volume-average diameters were controlled approximately at 30 p,m, 8 ~tm and 4 pm grades when cyclohexane, liquid paraffin and pump oil were used as dispersants, respectively. The present investigations reveal that higher viscosity dispersant is suitable for the preparation of smaller-size microspheres, while larger size microspheres are prepared preferably using lower-viscosity dispersant. The chiral stationary phase derived from 8 μm grade microspheres can separate the enantiomers of efavirenz.

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.

Determination of Doxycycline in Doxycycline Hydrochloride Soluble Powder under Different Chromatographic Conditions

[Objectives]This study was conducted to compare the accuracy and detection speed of high performance liquid chromatography for determining the content of doxycycline hydrochloride in doxycycline hydrochloride soluble powder under different chromatographic conditions, in order to improve the level of laboratory testing. [Methods] Four sets of experiments were designed through a cross-test method. The contents of doxycycline hydrochloride in the raw materials of doxycycline hydrochloride and doxycycline hydrochloride soluble powder were determined by changing the chromatographic column and mobile phase conditions, and the feasibility and practicability of the four methods were judged by comparing the detection results through data and chromatographic signal processing. [Results] The content of doxycycline hydrochloride in finished doxycycline hydrochloride soluble powder and the raw materials of doxycycline hydrochloride could be accurately determined under the four chromatographic conditions, of which the experimental group using Agilent C18 column and the mobile chromatography conditions of finished doxycycline hydrochloride soluble powder had the shortest retention time. [Conclusions] The high performance liquid chromatography method using Agilent C18 chromatographic column and mobile phase chromatographic conditions of finished doxycycline hydrochloride soluble powder products can quickly and accurately determine the content of doxycycline hydrochloride in doxycycline hydrochloride soluble powder.

通腑泻下颗粒指纹图谱的建立及7种成分的定量分析

建立通腑泻下颗粒(TongFu XieXia Granules,TFXXG)的高效液相色谱(HPLC)指纹图谱,并同时测定大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、厚朴酚以及和厚朴酚的含量。采用Waters Symmetry C18(2.1 mm×150 mm,3.5μm)色谱柱,以0.1%磷酸水溶液(A)和甲醇(B)为流动相梯度洗脱;流速为0.4 mL/min;柱温为30℃;检测波长为254 nm;进样量为10μL。建立10批TFXXG的指纹图谱,并采用“中药色谱指纹图谱相似度评价系统”(2012版)计算相似度;测定大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、厚朴酚以及和厚朴酚的含量。结果表明,10批TFXXG共确定出25个共有峰,相似度均在0.950以上;共指认出12个共有峰,分别为芥子碱硫氰酸盐、咖啡酸、松果菊苷、柚皮苷、新橙皮苷、芦荟大黄素、大黄酸、厚朴酚、和厚朴酚、大黄酚、大黄素以及大黄素甲醚。其中,芦荟大黄素、大黄酸、厚朴酚、和厚朴酚、大黄素、大黄酚以及大黄素甲醚的含量分别为0.1244~0.1304、0.4271~0.4387、3.0479~3.0853、1.2910~1.3039、0.2427~0.2523、0.7718~0.7897和0.5085~0.5249 mg/g。本文所建指纹图谱和定量分析方法重复性好,可操作性强,可用于控制TFXXG的质量。

液相色谱仪计量性能期间核查

参照JJG 705—2014,对其中的部分方法进行优化,从泵流量设定值误差、流量稳定性、梯度误差、波长示值误差、波长重复性、基线噪声、基线漂移、最小检测浓度、线性范围、定性重复性、定量重复性11个方面对液相色谱仪进行系统核查。核查结果显示:泵流量设定值误差为0.5%,流量稳定性为0.8%,梯度误差为±0.2%,波长示值误差为1.0nm,波长重复性为1.2 nm,基线噪声为1.6×10^(-4),30 min内基线漂移为8×10^(-4),最小检测质量浓度为2.2×10^(-8)g/mL,线性范围为1578.9,定性重复性为0.5%,定量重复性为0.8%。该方法的实验数据可为液相色谱仪管理者提供仪器评价依据,为食品药品企业验证液相色谱仪提供参照依据。

纳米棒状MgO固相萃取结合高效液相色谱荧光法测定食用油中的苯并[a]芘

采用直接沉淀法合成了一种纳米棒状Mg O吸附剂,结合固相萃取-高效液相色谱荧光检测(SPE-HPLC/FLD)建立了食用油中苯并[a]芘(Ba P)的快速分析方法。采用X射线衍射(XRD)、傅里叶变换红外光谱(FTIR)、扫描电子显微镜(SEM)、比表面积和孔径分析仪(BET)对合成的棒状Mg O进行了表征,结果表明Mg O纳米材料是多晶立方结构,呈棒状形貌,具有较大的比表面积,非常适合作为SPE的吸附剂。对SPE过程中吸附剂用量、清洗液种类、解吸液种类及用量、上样液体积进行了详细的优化。在优化的条件下,食用油中Ba P在0.5~20.0μg/kg浓度范围内,线性方程回归系数的平方(R^(2))为0.9999,以信噪比的3倍和10倍计算方法检出限和定量限分别为0.10μg/kg和0.33μg/kg。同时,Ba P在低、中、高3种加标浓度下回收率在85.3%~96.5%,日内日间相对标准偏差范围在2.3%~9.0%,表明本方法具有很好的准确度和精密度。最后,将该方法应用于8种实际油样中Ba P的分析,所有样品的回收率在77.8%~92.5%之间,相对标准偏差小于9.8%,表明该方法具有很好的适用性,在实际样品分析中具有很好的应用前景。

高效液相-蒸发光散射检测器(HPLC-ELSD)测定红参中的精氨酸单糖苷(AF)及精氨酸双糖苷(AFG)含量

目的建立一种同时测定人参加工品中精氨酸单糖苷(AF)和精氨酸双糖苷(AFG)含量的方法。[方法]用PrevailTMC18column(4.6 mm×250 mm,5μm)色谱柱进行检测。流动相A:色谱乙腈,流动相B:5.0mmol/L七氟丁酸的0.7%三氟醋酸溶液。色谱条件为[0%A(0 min);0%A(5 min);15%A(8 min);35%A(25 min)];流速0.8 mL/min;漂移管温度为115℃;气体流量3.2 L/min。[结果]AF和AFG的检测限分别为0.015和0.010 mg/mL;线性关系相关系数分别为,AF:0.9997,AFG:0.9999,表明线性关系良好。AF和AFG检测的精密度,重复性,稳定性以及回收率的相对标准偏差分别为0.43%&0.37%,0.43%&0.55%,0.43%&0.49%,0.45%&0.15%。AF和AFG检测的回收率分别为99.5%&100%,表明方法稳定。经检测,高丽红参,中国红参和生晒参中AF含量分别为0.74%,0.91%和1.14%,AFG含量分别为6.69%,5.12%和0.85%。[结论]这种方法成功用于高丽红参,中国红参及生晒参中AF及AFG的检测;且红参中AF和AFG含量明显高于生晒参。该方法测定结果可靠,缩短了测定时间,较少杂质干扰。但因为本研究考察氨基酸种类较少,当衍生物种类较多时,仍需进一步考察其分离度。

柱前衍生法-HPLC测定化妆品中甲醛的不确定度评定

分析柱前衍生法-HPLC测定化妆品中甲醛的不确定度。根据《化妆品安全技术规范》(2015年版)第四章理化检验4.12方法甲醛测定和JJF1059.1-2012《测量不确定度评定与表示》,建立分析柱前衍生法-高效液相色谱法测定甲醛的不确定度数学模型,通过对整个测定过程中的不确定度来源进行分析和合成,运用最小二乘法对甲醛标准曲线拟合不确定度进行评定。取样量为0.2 g时,置信区间为95%时,K=2,化妆品中甲醛含量为(14.9±0.072104)mg/g。

加味增液汤HPLC指纹图谱及3种主要成分的含量测定

目的建立加味增液汤HPLC指纹图谱并对3种主要成分进行含量测定。方法色谱柱为岛津WondaSil C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.3%磷酸水溶液,梯度洗脱;流速为1.0 mL/min;检测波长为265、203、310、290 nm;柱温为25°C;进样量为20μL。建立10批加味增液汤HPLC指纹图谱,采用中药色谱指纹图谱相似度评价系统软件(2012A版)进行相似度分析,对共有峰进行鉴定和药材归属,并测定3种主要成分的含量。结果10批加味增液汤的指纹图谱有17个共有峰,其相似度介于0.872~0.989之间。指认出9、14、17号峰分别为阿魏酸、橙皮苷、哈巴俄苷。阿魏酸、橙皮苷、哈巴俄苷的含量分别为0.0673~0.1748、0.4988~1.5227、0.2709~0.8024 mg/g,转移率分别为30.74%~55.63%、11.77%~35.94%、23.15%~68.56%。结论该研究所建立的HPLC指纹图谱结合主要成分定量分析方法可用于加味增液汤汤剂的质量分析与控制,分析方法简单,结果可靠。

基于UPLC-Q TOF-MSE技术的藤黄健骨丸指纹图谱建立及共有峰鉴定

建立藤黄健骨丸高效液相色谱(HPLC)指纹图谱,完善产品质量标准。采用Waters Symmetry C18(150 mm×2.1 mm,3.5μm)色谱柱;流速为0.4 mL/min;进样量为2μL;紫外检测波长为260 nm;在柱温30℃下以0.1%甲酸水(A)和0.1%甲酸乙腈(B)为流动相进行梯度洗脱。利用“中药色谱指纹图谱相似度评价系统(2012版本)”建立藤黄健骨丸指纹图谱,并分析相似度。采用液质联用技术(UPLC-Q TOF-MSE),根据化合物串联质谱信息结合对照品进行了共有峰结构鉴定。共确定了10批藤黄健骨丸指纹图谱中20个共有峰,相似度均在0.95以上。基于对照品和串联质谱信息指认了共有峰中13种成分的结构,并进行了峰归属。对10批藤黄健骨丸指纹图谱共有峰的相对保留时间和相对峰面积进行了计算,结果显示各批次样品共有峰的相对保留时间和相对峰面积均比较稳定,表明它们所对应的各成分含量较为均一。采用该指纹图谱对中间体进行了相似度评价,发现中间体与成品相关性良好。该方法精密度、重复性和稳定性好,不仅可用于藤黄健骨丸成品的质量控制,还可对中间体进行检测,用于生产过程的控制。

亲脂性匹配色谱分离-超高效液相色谱-串联质谱法测定果蔬中21种三唑类杀菌剂

为了提高农药残留的色谱分离效率和降低基质效应,本研究提出亲脂性匹配色谱分离,选取三唑类杀菌剂为研究对象,建立了果蔬中21种三唑类杀菌剂的超高效液相色谱-串联质谱(Ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)检测方法。样品经乙腈提取,盐析分相,分散固相萃取净化,选用与三唑类杀菌剂具有相近亲脂性的色谱柱进行分离,探究不同亲脂性烷基键合相对基质效应、回收率以及三唑杀菌剂和基质组分色谱分离的影响。结果表明,亲脂性匹配色谱分离能够提高色谱分离效率,改善三唑杀菌剂和基质组分的色谱分离,21种三唑杀菌剂的基质效应为-8.3%~4.7%,在5~250μg/L浓度范围内线性关系良好,决定系数R2≥0.999,平均回收率为91.4%~108.1%,定量限为0.5~3.5μg/kg。该检测方法能够有效降低基质效应,使用溶剂校准曲线进行定量即可获得满意的回收率,显著提高了检测效率,具有简便、准确、灵敏度高等特点,适用于果蔬中三唑杀菌剂的检测。所述亲脂性匹配色谱分离,为农药残留的液相色谱-串联质谱法分析中液相色谱柱的选择和基质效应的降低提供了方法参考。

参苏丸质量标准提高研究

目的通过修订参苏丸中甘草的鉴别方法,并整合参苏丸陈皮中橙皮苷、枳壳中柚皮苷和新橙皮苷的含量测定方法,提高参苏丸质量标准。方法在2020年版《中国药典》方法的基础上,对参苏丸中甘草的薄层色谱鉴别方法进行了改进,同时新增高效液相色谱法对陈皮中橙皮苷、枳壳中柚皮苷和新橙皮苷的含量测定。结果改进薄层色谱法可以用于鉴别处方中的甘草。含量测定方法柚皮苷在0.0140~0.2235mg·ml^(-1)(r=1.0000)范围内有良好的线性关系,水丸平均加样回收率为97.22%,RSD为0.68%(n=6),蜜丸平均加样回收率为99.43%,RSD为0.32%(n=6);橙皮苷在0.0114~0.1824mg·ml^(-1)(r=1.0000)范围内有良好的线性关系,水丸平均加样回收率为95.93%,RSD为0.48%(n=6),蜜丸平均加样回收率为96.86%,RSD为0.32%(n=6);新橙皮苷在0.0131~0.2102mg·ml^(-1)(r=1.0000)范围内有良好的线性关系,水丸平均加样回收率为100.21%,RSD为0.49%(n=6),蜜丸平均加样回收率为97.57%,RSD为0.42%(n=6)。结论本方法专属性强、重现性好,可有效控制参苏丸质量。

QuEChERS-高效液相色谱法测定食品中溴硝醇残留量

目的 建立QuEChERS-高效液相色谱法测定食品中溴硝醇残留量的分析方法 。方法 前处理采用QuEChERS方法 ,样品粉碎后用甲醇提取,改性多壁碳纳米管净化, Agilent ZORBAX Eclipse XDB-C18色谱柱(250 mm×4.6 mm, 5μm)对净化液进行分离,以甲醇-乙腈(含0.1%磷酸)体系作为流动相, 210 nm波长下检测分析。结果 在优化实验条件下,溴硝醇在0.02~10.00μg/mL范围内具有良好的线性关系,相关系数大于0.9999,检出限(S/N=3)为0.050 mg/kg,加标回收率为95.3%~104.0%,相对标准偏差为4.2%~5.3%,实际样品检测中均未检出溴硝醇。结论 该方法 简单、快速、准确、灵敏度高,可应用于粮食、水果和蔬菜中溴硝醇残留的快速检测。

现代分析技术在食品分析检测过程中的应用

食品分析与检测是确保食品安全性、营养价值和合规性的关键过程,涵盖营养成分、添加剂和污染物的识别与量化。本文探讨了食品分析的基本原理,包括样品采集、处理和不同分析方法,以及现代分析技术如高效液相色谱技术、气相色谱技术、质谱技术等在食品安全评估和质量控制中的应用,旨在提高食品分析检测的灵敏度和准确性,维护食品安全和提高公众健康水平。

复方醋酸氟轻松酊有关物质测定方法研究

目的:建立复方醋酸氟轻松酊中有关物质的检测方法。方法:色谱柱为Ultimate LP-C18(250*4.6mm,5μm),流动相为55%甲醇水溶液,流速为1.0 mL/min,检测波长为235nm和245nm,柱温为30℃,进样量为10μL。结果:专属性试验中,已知杂质与主峰及杂质与杂质峰之间分离度大于2.0,各峰间分离良好;系统适用性试验中,峰面积RSD值为1.42%,保留时间RSD值为0.84%;溶液稳定性试验中,醋酸氟轻松原料药溶液24h内杂质极差变化小于0.05%;精密度试验中,氟轻松测定12次的结果极差在0.1%以内;定量限和检测限试验中,氟轻松定量限浓度为供试品浓度(0.4mg/mL)0.032%;线性与范围试验中,醋酸氟轻松在0.30μg/mL~6.04μg/mL范围内线性关系良好,方程:y=16807 x+3692.8;准确度试验中,氟轻松回收率在80.0%~120.0%,平均回收率为100.5%,RSD为1.5%;耐用性试验中,改变条件后测得单杂和总杂含量变化在0.1%之间。结论:该方法专属性强、灵敏度好、准确度高,可用于复方醋酸氟轻松酊中有关物质的检测。