免疫亲和柱净化、在线电化学衍生化高效液相色谱法检测花生中的黄曲霉毒素

 采用免疫亲和柱净化、在线电化学衍生化高效液相色谱法测定了花生中黄曲霉毒素（AFT）B 1,B2,G1和G2。以体积分数为80%的甲醇提取样品中的AFT,经免疫亲和柱净化洗脱 后,以Kobra Cell装置在线衍生,反相HPLC分离定量。4种毒素的分离在13 min内完成,检 出限均达到0.1 μg/kg。5次测定花生样品的RSD值为9.2%～15%;样品添加标样0.5 ～9.0 μg/kg,回收率为74.8%～97.3%。     

免疫亲和-光化学衍生高效液相色谱检测花生及花生制品中黄曲霉毒素

建立了免疫亲和柱净化、柱后光化学衍生、高效液相色谱法结合荧光检测器测定花生及花生制品中黄曲霉毒素的新方法。方法的检出限为0.25μg/kg,相对标准偏差低于10%,回收率为64%~101%。该方法简便快速、灵敏准确、重复性好,满足出入境检验检疫工作的需要。     

超高效液相色谱法快速检测粮食中黄曲霉毒素的含量

建立了免疫亲和柱净化-超高效液相色谱法快速测定粮食中黄曲霉毒素(Aflatoxins,AF)的检测方法.样品经提取后,用免疫亲和柱净化、浓缩,Waters Acquity UPLC BEH C18色谱柱(50 mm×2.1 mm,1.7 um)分离.以甲醇-水(40∶60,V/V)为流动相,流速为0.2 mL/min,进样量为1μL,荧光检测器检测,激发波长为360 nm,发射波长为440 nm,无需衍生.黄曲霉毒素B1,B2,G1,G2的保留时间小于5min,从样品前处理到结果分析整个过程小于45 min.根据3倍信噪比的峰响应值,确定黄曲霉毒素(B1,B2,G1,G2)检出限分别为0.15,0.05,0.40,0.06 pg,4种毒素在0.4 ～ 60.0 pg,0.2～15.0 pg,1.5～ 60.0 pg和0.2～15.0 pg范围内分别呈线性相关,相关系数R2值分别为0.9999,0.9999,0.9998和0.9992;在小麦、玉米、稻谷3类样品中加标回收率为77.4％ ～ 104.2％,精密度为1.8％ ～ 8.9％.本方法无需衍生即可同时测定粮食中4种黄曲霉毒素,适用于粮食中黄曲霉毒素的快速定量测定.     

液相色谱-电化学衍生-光学检测技术研究进展

综述了以反相键合相为主的高效液相色谱-柱后电化学衍生-荧光检测(RP-HPLC/ED/FD)技术的研究进展。对RP-HPLC/ED/FD存在的主要问题进行了讨论。并介绍了离子色谱-柱后电化学衍生-光学联用装置的构建及其在极性有机物和金属离子的测定方面的应用研究。     

多组分免疫亲和柱净化/超高效液相色谱-串联质谱检测水产饲料中的黄曲霉毒素

建立了多组分免疫亲和柱净化/超高效液相色谱-串联质谱(UPLC-MS/MS)测定水产饲料中黄曲霉毒素AFB_1、AFB_2、AFG_1及AFG_2含量的方法。饲料样品用80%乙腈水超声提取后,经多组分免疫亲和柱净化,采用UPLC-MS/MS测定,外标法定量。以0.1%甲酸-乙腈为流动相,梯度洗脱分离,电喷雾正离子多反应监测模式检测。结果表明,AFB_1、AFG_1和AFB_2、AFG_2分别在2.25～22.5 ng/mL和0.75～7.5 ng/mL质量浓度范围内呈良好线性,相关系数大于0.997;AFB_1、AFG_1的定量下限均为0.7μg/kg,AFB_2、AFG_2的定量下限均为0.2μg/kg。AFB_1、AFG_1在1.5μg/kg,AFB_2、AFG_2在0.5μg/kg加标水平下的回收率为78.7%～85.5%,日内相对标准偏差(RSD)为6.0%～6.5%,日间RSD为6.6%～7.0%。该法操作简单,耗时少,重复性好,灵敏度高,适用于水产饲料中黄曲霉毒素的测定。     

荧光色谱-柱后光化学衍生法测定食品中的黄曲霉毒素B1

采用免疫亲和层析柱吸附食品样品中的黄曲霉毒素B1,以荧光色谱-柱后光化学衍生法测定食品中黄曲霉毒素B1的含量。样品用甲醇-水溶液（体积比为7：3）混合,均质器高速搅拌提取,经免疫亲和层析柱纯化后,通过LachromC18色谱柱进行分离,以甲醇一水溶液（体积比为45：55）作为流动相,荧光检测器激发波长为360nm、发射波长为420nm,外标法定量。黄曲霉毒素B,的质量浓度在1～20ng／mL范围内与色谱峰面积线性关系良好,相关系数r=0．9998,检出限为0．1gg／kg。在3个添加水平下,样品的加标回收率在88．5％～97．9％之间,相对标准偏差均小于3％（n=6）。该方法操作简单、定量准确,可用于食品中黄曲霉毒素B．的定量定性检验。     

高效液相色谱串联质谱法检测腰果中黄曲霉毒素

建立了腰果中4种黄曲霉毒素的高效液相色谱-串联质谱检测方法(HPLC-MS/MS)。样品用甲醇-水(8:2, v/v)溶液提取后用弗罗里硅土柱净化,5 mL丙酮-水-甲酸溶液(96:3.5:0.5, v/v/v)洗脱,氮吹至干,1 mL甲醇定容;在资生堂MG C18色谱柱(100 mm×3.0 mm, 3 μm)上梯度洗脱分离,然后采用电喷雾离子化三重四极杆串联质谱测定。实验结果表明,4种黄曲霉毒素在各自的线性范围内峰面积与其质量浓度线性关系良好,相关系数(r2)大于0.997;检出限(信噪比为3)为0.009～0.04 μg/kg,定量限(信噪比为10)为0.03～0.12 μg/kg;平均回收率为63.0%～78.5%,相对标准偏差为2.8%～9.1%,均符合痕量分析的要求。评价了基质效应,信号抑制/增强值为88.8%～99.4%,说明净化后的基质效应较小。该方法简单快速、准确可靠,可用于腰果中黄曲霉毒素的检测。     

高效液相色谱-串联质谱法检测花生中的黄曲霉毒素B_1

应用高效液相色谱-电喷雾串联四极杆质谱联用系统(HPLC-MS/MS),在多反应离子检测方式(MRM)下,对花生中的黄曲霉毒素B1进行检测.对花生中黄曲霉毒素B1的提取、净化、液相分离及串联质谱等相关检测参数进行了优化研究.用V(甲醇):V(水)=6:4提取,OASIS HLB SPE小柱净化,定容过滤.采用V(甲醇):V(水)(含体积分数0.1%甲酸)=7:3为流动相,前级离子313.0,二级离子241.1、269.1,ESI正离子方式检测,在3.3 min出峰.结果表明,在ESI正离子模式下,黄曲霉毒素B1在其线性定量范围0.1～50μg/kg内,相关系数达到0.9999,检出限为0.03μg/kg,最低定量限为0.1μg/kg.低、中、高浓度添加回收率范围为93%～105%.     

高效液相色谱法测定动物组织样品中黄曲霉毒素的残留量

采用免疫亲合技术,对动物肝脏和肌肉中黄曲霉毒素残留进行了提取和纯化处理,建立了一种快速、灵敏、简便的测定动物组织中黄曲霉毒素B1、B2、G1、G2的高效液相色谱方法。用反相HPLC分离,柱后光化学衍生,荧光检测器测定黄曲霉毒素含量,保留时间定性,峰面积定量,测定了样品和标准样。结果表明,4种毒素可在10min内完成分离;线性范围为15～1500pg;线性回归系数大于0．9998。用本方法对动物肝脏和肌肉样品进行了加标回收实验,4种黄曲霉毒素的平均回收率为68．71％～83．42％;相对标准偏差为3．51％～7．40％;检出限均达到2．65pg。     

超声萃取-免疫亲和柱净化-柱后光化学衍生高效液相色谱法同时测定蜂房药材中4种黄曲霉毒素

建立超声萃取-免疫亲和柱净化-柱后光化学衍生高效液相色谱同时测定蜂房药材中黄曲霉毒素B1、黄曲霉毒素B2、黄曲霉毒素G1、黄曲霉毒素G2含量的分析方法。样品经粉碎,过孔径为120μm筛后,采用70%甲醇溶液超声处理30 min,经免疫亲和柱净化、高效液相色谱分离、光化学柱后衍生,通过荧光检测器测定4种黄曲霉毒素的含量。黄曲霉毒素B1的线性范围为0.010 4～0.052 0 ng,相关系数为0.999 9;黄曲霉毒素B2的线性范围为0.003 8～0.019 0 ng,相关系数为0.999 8;黄曲霉毒素G1的线性范围为0.010 8～0.054 0 ng,相关系数为0.999 8;黄曲霉毒素G2的线性范围为0.003 8～0.019 0 ng,相关系数为0.999 8。4种黄曲霉毒素检出限分别为0.42、0.15、0.43、0.15μg/kg,测定结果的相对标准偏差不大于2.5%(n=6),样品加标回收率为92.9%～96.9%。该方法操作简便,灵敏度高,可用于蜂房中黄曲霉毒素含量的测定。     

柱前衍生化-HPLC测定仲丁胺对映体的纯度

用3, 5-二硝基苯甲酸对仲丁胺进行柱前衍生, 将其衍生物在CHIRALCEL OD-H手性固定相上拆分, 并通过二极管阵列紫外检测器对其衍生物进行检测, 建立了一种拆分仲丁胺消旋体、测定仲丁胺光学纯度的新方法. 以正己烷和乙醇或异丙醇为流动相, 在CHIRALCEL OD-H手性固定相上对仲丁胺衍生物进行了拆分, 并考察了流动相组成和柱温对该对映体衍生物分离的影响, 获得较好的分析条件, 分离因子大于1.2. 非手性试剂柱前衍生化法测定仲丁胺光学纯度与旋光度方法比较, 结果一致. 方法可用于仲丁胺对映体的质量控制.     

柱后衍生高效液相色谱法测定鸡肉中莫能菌素残留量

样品用甲醇－水溶液匀浆、提取后过滤,再用二氯甲烷进行液－液萃取,然后通过Ｓｅｐ－Ｐａｋ柱进一步净化。净化后的样液与香兰素衍生试剂在酸性和加热条件下进行柱后衍生,反应产物在５２０ｎｍ波长处测定。衍生剂：香兰素３０．０ｇ＋浓硫酸２０ｍＬ＋甲醇９５０ｍＬ,流速：０．７ｍＬ／ｍｉｎ;色谱柱：μ－ＢｏｎｄａｐａｋＴＭＣ１８３．９ｍｍｉ．ｄ．×３００ｍｍ,或相当的色谱柱;流动相：Ｖ（甲醇）Ｖ（水）Ｖ（磷酸）＝９４０６０１,流速０．７ｍＬ／ｍｉｎ;柱后反应室：体积大于１．４ｍＬ的不锈钢管（３００ｃｍ×１ｍｍｉ．ｄ．）。     

柱后光化学衍生高效液相色谱法同时测定牛奶中黄曲霉毒素

建立了高效液相色谱法同时测定牛奶中黄曲霉毒素B1,B2,G1,G2的方法。用乙腈和水的混合溶液(体积比为80∶20)提取牛奶样品中4种黄曲霉毒素,提取液经Mycosep 228 AflaPat多功能净化柱净化,浓缩后采用C18色谱柱分离,光化学衍生后进入荧光检测器测定,外标法定量。对牛奶样品进行加标回收和精密度试验,黄曲霉毒素B1,B2,G1,G2的检出限分别为0.50,0.10,0.50,0.10μg/kg,回收率均在85%以上,测定结果的相对标准偏差为1.72%～3.52%(n=6)。该方法操作简单,速度快,重现性好,满足牛奶中黄曲霉毒素检测的要求。     

柱前衍生高效液相色谱法测定啤酒中微量甲醛

甲醛作为消毒剂、工业助剂等已广泛应用于各行各业,也列在食品工业加工助剂推荐名单中[1]。然而,甲醛能引起头痛、恶心,肠胃不舒服、皮肤过敏等反应。美国环保署(EPA)称甲醛可能是致癌诱导物[2]。我国居室内空气中甲醛卫生标准不大于0·08mg/m3,而饮用水和饮料类中甲醛允许含量     

硝基氟苯柱前衍生HPLC测定减肥药中芬氟拉明

诺贝尔奖获得者F．sange使用2,4-二硝基氟苯(DNFB)确定了蛋白质中氨基酸排列次序以来,DNFB常用作为胺类化合物的衍生试剂。在碱性溶液中,DNFB与胺反应生成强紫外吸收的硝基苯胺类化合物。芬氟拉明(Fenfluramine,FFA)化学名为N-乙基-α-甲基-3-三氟甲基苯乙胺,属苯乙胺类食欲抑制剂,对人体健康有潜在危害。HPLC分析FFA的报道,检测限仅为20．6ng。本文用DNFB为衍生试剂,研究衍生化条件,色谱分析检测限降至1．02ng。     

Determination of sugars by liquid chromatography with post-column catalytic derivatization and fluorescence detection

Summary A method for the determination of reducing sugars such as fructose and glucose and nonreducing sugar such as sucrose by high performance liquid chromatography followed by an acidic hydrolysis and a derivatization with benzamidine has been developed. After separation of sugars on a gel column packed with a polymer-based cation exchange material (Sugar-Pak I, Waters-Millipore), the sucrose is first hydrolysed in a solid phase reactor to convert it into reducing subunits. A post-column fluorigenic reaction with benzamidine under alkaline condition allows the selective determination of both natural and converted reducing carbohydrates.This procedure has proven to be selective (fluorigenic detection) and highly sensitive (allowing detection as little as picomoles amounts), reproducible and linear over a broad range of concentrations: 5×10^–4 to 1.0×10^–2 M.The applicability of this method to natural matrices such as plant extracts and beverages is also described. The sugar content of a barley extract has been determined and compared with a specific enzymatic test. The determined sugar content of natural and commercial lemon juices as well as of Cola beverages has been compared with those found by the conventional LC refractive index analytical procedure. In all cases, the results were comparable and were within the experimental errors of the methods.     

Determination of the impurity of isopentylamine hydrochloride in the intermediate of bortezomib by HPLC with pre-column derivatizationEI北大核心CSCD

A method was developed for the simultaneous determination of （R）-pinacol-1-amino-3-methylbutylborate hydrochloride and isopentylamine hydrochloride by pre-column derivatization high performance liquid chromatography （HPLC） with dichloromethane as solvent,triethylamine as acid binding agent and benzoyl chloride as derivative reagent. （R）-pinacol-1-amino-3-methylbutylborate hydrochloride and isopentylamine hydrochloride reacted with derivatization reagent in an ice bath for 10 min. The derivatives were qualitatively determined by HPLC-mass spectrometry（HPLC-MS）. HPLC was performed on an AgelaVenusil MP C18 （2）column using methanol-water （70∶30,V/V）as mobile phase. The flow rate was 1.0 mL/min,the detection wavelength was 236 nm and the column temperature was 35℃ . The UV absorption produced after derivatization,which could be detected by HPLC with good specificity. Isopentylamine hydrochloride derivative showed an excellent linearity in the range of 0.075-30 μµg/mL,with the limit of quantification and the limit of detection（LOD）of 0.075 and 0.030 μµg/mL,respectively. And the S/N of the LOQ and the LOD were 44.6 and 7.8,respectively. The average recoveries were 97.7%-104.0% with the RSD of 2.9%. The method is suitable for the determination of isoamylamine hydrochloride and has significant reference value for the determination of other amine salt without UV absorption. © 2022, Youke Publishing Co.,Ltd. All rights reserved.     

柱前衍生高效液相色谱法测定鱼罐头中的组胺

建立了一种测定鱼罐头中组胺含量的柱前衍生高效液相色谱(HPLC)方法。样品匀浆后采用高氯酸水溶液超声提取,提取液经丹酰氯衍生后,采用HPLC分离,紫外检测器检测,外标法定量。采用粒径为1.8 μm固定相填料的C18色谱柱,在0.3 mL/min的流速下,样品的分析时间小于5 min,并可有效地减少流动相消耗,节约成本。组胺在0.08～8.00 mg/L内线性关系良好,相关系数为0.99998;酱煮鲐鱼罐头中组胺在不同浓度水平的平均加标回收率均大于96%,相对标准偏差(RSD)小于2.5%;鱼罐头中组胺的定量限可达5.00 mg/kg。所建立的HPLC方法快速、灵敏度高、重复性好,前处理方法简单,可用于鱼罐头中组胺的测定。     

Determination and dynamics of kanamycin A residue in soil by HPLC with SPE and precolumn derivatization

As one of the aminoglycoside antibiotics, kanamycin has been widely used in human therapy and as an additive to promote growth and prevent disease in forage. The kanamycin residue may have potenital risks for organisms and the environment. Therefore, the monitoring of this drug may have dynamic importance. In this work, a novel method for determination and dynamic study of kanamycin A was developed through solid phase extraction and derivatization with 4-chloro-3,5-dinitrobenzotrifluoride before high-performance liquid chromatography analysis. The calculated recoveries were from 72.3 to 92.5%, with relative standard deviations from 2.99 to 6.94%. The detection limit of kanamycin A in soil was 0.006?mg?kg^?1 with a signal-to-noise ratio of 3. The dynamics in soil showed that the degradation of kanamycin A coincided with the equations C?=?1.951e^{?0.0482 t} for black soil and C?=?1.807?e^{?0.0247 t} for red soil and the half-lives were 14.38 and 28.06?d respectively. The degradation rate reached 95.19% in black soil after 63 days compared with 77.14% in red soil.     

高效液相色谱法测定茶叶中茶氨酸的研究

研究了用高效液相色谱法分析茶叶中茶氨酸的方法。以异硫氰酸苯酯（ＰＩＴＣ）作衍生剂、醋酸钠缓冲液与乙腈梯度洗脱、Ｃ１８柱分离、紫外２４３ｎｍ检测,探讨了茶氨酸的提取方法。比较了不同茶叶中茶氨酸的含量。方法具有快速、精确、灵敏等特点。