Modernization of Chinese herbal compound and the high performance liquid chromatography tandem mass spectrometry (HPLC-MS)

Chinese herbal compound is playing an important role on curing human diseases.And it has been a trend that Chinese herbal compound is being used all over the world in 21 century.However,our Chinese herbal compound is facing serious challenge for the lack of canonical system of quality criterion for Chinese herbal compound so it has been a urgent problem to set up the quality control standards and reveal therapeutic basis of Chinese herbal compound.In order to give full play to the advantages of Chinese herbal compound,modern scientific and technological is used to research of Chinese herbal compound,especially the high performance liquid chromatography tandem mass spectrometry(HPLC-MS),because it is high sensitive,rapid,and obtain more information.It is very necessary that HPLC-MS is uesed to elucidate the effective components of basic substances of Chinese Herbal Compound,and endow traditional Chinese medicine with modern scientific connotation.

Detection of 36 antibiotics in coastal waters using high performance liquid chromatography-tandem mass spectrometry

Among pharmaceuticals and personal care products released into the aquatic environment, antibiotics are of particular concern, because of their ubiquity and health effects. Although scientists have recently paid more attention to the threat of antibiotics to coastal ecosystems, researchers have often focused on relatively few antibiotics, because of the absence of suitable analytical methods. We have therefore developed a method for the rapid detection of 36 antibiotic residues in coastal waters, including tetracyclines (TCs), sulfanilamides (SAs), and quinolones (QLs). The method consists of solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, using electrospray ionization (ESI) in positive mode. The SPE was performed with Oasis HLB and Oasis MCX cartridges. Chromatographic separation on a C18 column was achieved using a binary eluent containing methanol and water with 0.1% formic acid. Typical recoveries of the analytes ranged from 67.4% to 109.3% at a fortification level of 100 ng/L. The precision of the method, calculated as relative standard deviation (RSD), was below 14.6% for all the compounds. The limits of detection (LODs) varied from 0.45 pg to 7.97 pg. The method was applied to determine the target analytes in coastal waters of the Yellow Sea in Liaoning, China. Among the tested antibiotics, 31 were found in coastal waters, with their concentrations between the LOD and 212.5 ng/L. These data indicate that this method is valid for analysis of antibiotics in coastal waters. The study first reports such a large number of antibiotics along the Yellow Sea coast of Liaoning, and should facilitate future comprehensive evaluation of antibiotics in coastal ecosystems.

Determination of thyreostats in bovine urine using ultra-high performance liquid chromatography-tandem mass spectrometry

Five thyreostats(TSs),namely tapazole,thiouracil,methylthiouracil,propylthiouracil,and phenylthiouracil,were determined in bovine urine using ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)in positive electrospray ionization mode.Extraction and clean-up were achieved using a ChemElut cartridge with tert-butyl methyl ether,without a derivatization step.Separation was achieved on an Acquity UPLC SS T3 column.The mobile phase was acetonitrile and water containing 0.2%(v/v)formic acid.The mass spectrometer was operated in multiple reaction monitoring mode.Urine samples were spiked with TS solution at levels corresponding to 5,10,15,and 20μg/L.The accuracy(internal standard corrected)ranged from 92%to 107%,with a repeatability precision(relative standard deviation,RSD)less than 15%for all five analytes.The RSDs within-laboratory reproducibility was less than 26%.The decision limits(CCα)and detection capabilities(CCβ)were obtained from a calibration curve and were in the ranges of 3.1-6.1μg/L and 4.0-7.4μg/L,respectively.The CCαand CCβvalues were below the recommended concentration,which was set at 10μg/L.The results show that the described method is suitable for the direct detection of TSs in bovine urine.This method can also be used to determine TSs in porcine urine.

Determination of okadaic acid related toxins from shellfish (<i>sinonovacula constricta</i>) by high performance liquid chromatography tandem mass spectrometry

Consumption of shellfish contaminated with algal toxins produced by marine dinoflagellates can lead to diarrhetic shellfish poisoning (DSP). It was therefore essential that there are analytical techniques to identify and quantify DSP toxins in shellfish. This new methodology could facilitate DSP monitoring and create a means of rapidly responding to incidents threatening public health. In the last years there were different analytical methods for DSP, such as mouse bioassay and LC-FLD. With the development of instrument, Liquid chromatography-mass spectrometry was substituted for other analytical methods with its good sensitivity and selectivity and without derivatization for the determination of DSP. In this report, a high performance liquid chromatogra-phytandem mass spectrometric(HPLC-MS/MS)method was developed for the simultaneous determination of okadaic acid (OA) and dinophysistoxins(DTX1) in Sinonovacula constricta. Optimization of pretreatment experiment was carried out to maximize recoveries and the effectiveness. The analytes were determined under multi-reactions monitoring (MRM) scan type with tandem mass analyzer using negative ion electrospray ionization (-ESI) mode .Finally, the detection and identification of OA and DTX-1 were based upon their retention times (RT) and the fragmentation patterns of their mass spectra. The method of LOQ for the two poisons was 0.02 mg·kg-1.The real sample test showed that this method could be used for sensitive, fast, and accurate determination of the two diarrheic shellfish poisons in shellfish.

Determination of 24 Allergens in Perfume with High Performance Liquid Chromatography Tandem Mass Spectrometry

A method of 24 allergens determination in cosmetics were established with high performance liquid chromatography tandem mass spectrometry. The targeted compounds were extracted with acetonitrile and determined with LC-MS/MS (MRM mode) with external method. The linearity between concentrations and peak area ratio was obtained from 1.0~5.0 mg/L. The limits of detection were 1.0 mg/L for the instrument and 5.0 mg/kg for the method respectively. The LOQ was 15.0 mg/L. The average recoveries of 24 allergens were between 85.9% and 110.0% at spiked levels of 5, 10 and 20 mg/kg with relative standard derivation (RSDs) of 5.5%~12.0%(n=10). The method could be used as a reliable means for simultaneous quantitative determination of allergens in cosmetics.

HPLC及LC-MS测定榛子中氯吡苯脲和多菌灵残留

本文采用高效液相色谱法(HPLC)和液相色谱-质谱法(LC-MS)对干果榛子中氯吡苯脲和多菌灵残留量进行含量测定.HPLC和LC-MS均使用C18色谱柱,含量测定采取等度洗脱,流动相:甲醇-水(55∶45,体积比);体积流量:1.0 mL·min-1;测定波长:260 nm;柱温:30℃.LC-MS采取电喷雾离子源,梯度洗脱,体积流量:0.6 mL·min-1,柱温:30℃.结果表明,榛子中氯吡苯脲与多菌灵存在残留,氯吡苯脲质量浓度在1.00~10.00μg·mL-1范围内线性关系良好,加样回收率在95.58%~100.58%;多菌灵质量浓度在1.005~15.075μg·mL-1范围内具有良好的线性关系,加样回收率在95.61%~104.39%.实验证明,HPLC与LC-MS相结合的方法具有操作简便、灵敏度高、检出限低等优点,能有效地检测到榛子样品中膨大剂氯吡苯脲及杀菌剂多菌灵的残留,并确定其残留量,线性关系和回收率结果均令人满意.根据被检测的8批样品中氯吡苯脲和多菌灵两项农药残留量推断,作为一般干果食用榛子是安全的.

HPLC-MS/MS测定保健食品中6种大麻素

目的:利用高效液相色谱-串联三重四极杆质谱(HPLC-MS/MS)对保健食品中6种大麻素进行定性定量分析。方法:HLB固相萃取小柱净化,采用XBridge C18色谱柱,乙腈溶液作为有机相,10mmol·L^(-1)乙酸铵水溶液作为水相,2种离子模式同时扫描,同位素内标定量。结果:所测大麻素在1-100μg·L^(-1)浓度范围均呈现良好线性关系,相关系数(r2)均>0.999。方法的检出限和定量限分别为5μg·kg^(-1)和10μg·kg^(-1)。该方法稳定性好,灵敏度高,适用于常见保健食品中6种大麻素的定性定量分析。结论:本研究成功建立了保健食品中6种大麻素的定性定量分析,为保健食品品质评价、安全监管及风险防范提供技术支持。

高效液相色谱-质谱法(HPLC-MS)测定化妆品中达克罗宁的含量

建立了一种高效液相色谱-质谱法(HPLC-MS)测定化妆品中达克罗宁含量的方法。样品经乙腈提取15 min后,用体积分数20%乙腈溶液稀释,经Capcell PAK C_(18)色谱柱(100 mm×2.1 mm×2μm)分离后,多反应监测模式进行检测。结果表明:达克罗宁在1.0~25.0 ng/mL线性关系良好,相关系数(r)大于0.995;加标回收率在89.4%~104.5%。该方法高效、准确,可用于化妆品中达克罗宁的定性和定量检测。

HPLC-MS/MS法测定保健品中麻黄碱、伪麻黄碱的不确定度评估

建立HPLC-MS/MS法测定保健品中麻黄碱、伪麻黄碱的测定不确定评估方法。通过构建麻黄碱、伪麻黄碱含量的数学模型,对不确定来源进行分析。分析结果表明,在麻黄碱和伪麻黄碱测定过程中不确定度主要来源是标准溶液的配制和标准曲线的拟合。当保健品中麻黄碱的含量为9.850μg/kg时,其扩展不确定度为0.7458μg/kg(k=2);伪麻黄碱含量为9.557μg/kg时,其扩展不确定度为0.9547μg/kg(k=2)。

HPLC-MS/MS测定食糖中4种合成着色剂

目的:建立高效液相色谱-串联质谱法(High Performance Liquid Chromatography-Tandem Mass Spectrometry,HPLC-MS/MS)测定食糖中4种着色剂的分析方法。方法:样品经80%甲醇超声提取,再用PWA-2固相萃取柱净化后,经VP-ODS C_(18)柱(2.0 mm×150 mm,5.0μm)分离,以乙酸铵和甲醇为流动相,梯度洗脱,用HPLC-MS/MS检测。结果:4种化合物在考察范围内线性良好,相关系数均大于0.999;加标回收率在85.36%~102.30%,RSD为0.42%~2.72%;检出限为2~5μg·kg^(-1)。结论:该方法灵敏度高、分辨率高、分析速度快,适用于食糖中合成着色剂的快速筛查和定量分析。

QuEChERS-HPLC-MS/MS法检测蚕砂中氯霉素残留量

本文建立了一种QuEChERS和C18固相萃取柱净化结合液相色谱串联质谱方法测定蚕砂中氯霉素残留。经乙酸乙酯提取,GCB和C18固相萃取柱净化,以甲醇和水为流动相梯度洗脱,C18柱(2.1 mm×100 mm,1.7μm)分离,在电喷雾离子源(ESI)负离子模式下采用液相色谱-串联质谱仪测定,内标法定量。结果显示,在0.02μg/L~5.00μg/L,氯霉素具有良好的线性关系,相关系数为1.000。氯霉素在4个加标水平(0.1μg/kg、0.3μg/kg、1.0μg/kg、2.0μg/kg)下回收率范围为74.7%~103.0%,RSD在4.36%~9.64%。该方法操作快速简单,且灵敏度与回收率高,可用于蚕砂中氯霉素残留的检测分析。

UPLC-MS/MS法检测3种食品中松仁过敏原

基于食品基质中松仁过敏原Pin k 2建立了一种超高效液相色谱-串联质谱法。将松仁经过研磨、脱脂、浸提、酶解后经Easy-nLC 1000-QExactive高分辨质谱仪进行分离分析,结合Uniprot蛋白数据库以及ProteinPilotTM软件对质谱图进行数据处理,经BLAST验证特异性,最终筛选3条松仁特异性肽段。方法学验证结果表明,方法在0.001~50mg/mL范围内线性关系良好,定量限为1mg/kg;在饼干、巧克力和饮料3种空白基质中的平均回收率为88.50%~107.57%,相对标准偏差不高于6.08%,基质效应为89.77%~96.13%。该方法具有灵敏度高、特异性好的优势,可应用于饼干、巧克力、饮料等食品样品中松仁过敏原的检测,为我国食品标签真实性检验及食品中隐性过敏原的检测提供技术支持。

COFs材料磁固相萃取-HPLC-MS/MS法测定烘焙食品中两种主要晚期糖基化终末产物

为实现烘焙食品中晚期糖基化终末产物(Advanced Glycation End Products,AGEs)的准确、高灵敏测定,建立基于磁性共价有机骨架(Covalent Organic Frameworks,COFs)材料的磁固相萃取(Magnetic Solid Phase Extraction,MSPE)技术,实现烘焙食品中两种主要AGEs,即羧甲基赖氨酸(N^(ε)-(carboxymethyl)lysine),(CML)和羧乙基赖氨酸(N^(ε)-(carboxyethyl)lysine,CEL)的选择性、高效富集,并结合高效液相色谱-串联质谱法对其进行定量分析。结果表明:所合成的磁性COFs材料具有较大的比表面积和较高的磁化强度,适合应用于MSPE技术。MSPE的适宜条件为吸附剂用量20 mg,涡旋萃取时间20 min,洗脱溶剂选用5%氨水/甲醇溶液,洗脱溶剂体积4 mL,洗脱时间15 min。CML和CEL的检出限分别为2.05 ng/mL和2.31 ng/mL,定量限分别为6.83 ng/mL和7.71 ng/mL;日内及日间精密度良好(RSD_(日内)≤4.74%,RSD_(日间)≤5.38%);在饼干、坚果2种典型样品基质中,CML和CEL的加标回收率分别为90.70%~108.74%、85.50%~113.00%。将本文所建立的分析方法应用于市售的26种烘焙食品中CML和CEL的含量测定发现,饼干、糕点类烘焙食品中二者含量分别为(9.60±0.16)~(78.10±1.12)mg/kg、(4.37±0.23)~(26.70±1.27)mg/kg;坚果类烘焙食品中二者含量分别为(6.49±0.13)~(82.00±0.98)mg/kg、(6.65±0.30)~(55.90±0.67)mg/kg,该方法能有效净化样品,显著降低基质效应,定量分析准确性较高,且所合成的磁性COFs材料可再生使用5次,适合烘焙食品中CML和CEL的检测。

QuEChERS-UPLC-MS/MS同时测定畜禽肉中23种驱虫类药物残留量

研究旨在建立同时检测畜禽肉中多种驱虫类药物的分析方法,通过优化前处理方法和仪器分析条件,建立了分散固相萃取法-超高效液相色谱-串联质谱法(UPLC-MS/MS)同时测定畜禽肉中23种驱虫类药物残留的分析方法,目标物以乙腈提取,分散固相萃取法(QuEChERS)净化,用乙腈+0.1%甲酸水作为流动相、UPLC-MS/MS确证检测、外标法定量。结果显示:所建立的方法定量限为0.2~1.0μg/kg,回收率为71.4%~107.5%,相对标准偏差为0.9%~8.8%。表明该方法定量限低,稳定性好,适用于畜禽肉中23种驱虫类药物残留的同时检测。

羊肉组织中6种抗微生物药物残留检测的HPLC-MS/MS方法建立

【目的】为了对羊肉及其副产品中抗微生物药物残留进行快速痕量检测,优化检测技术,建立羊可食组织中恩诺沙星、环丙沙星、氧氟沙星、庆大霉素、替米考星、泰乐菌素6种药物残留检测的高效液相色谱串联质谱法(HPLC-MS/MS)。【方法】对0.1%甲酸乙腈和0.1%甲酸甲醇2种不同药物提取液进行了对比;以动物性食品中氟喹诺酮类、氨基糖苷类和大环内酯类药物检测国家标准为参考,比较HLB和C_(18)2种固相萃取柱对6种目标药物的净化效果;同时,分别用0.1%甲酸水、乙腈、0.1%甲酸乙腈、甲醇、0.1%甲酸甲醇作为洗脱液对提取净化后的样品进行洗脱,对比交叉组合的提取和洗脱效果。将建立方法应用于实际样品检测,并与国家标准检测方法进行对比和统计分析。【结果】确定了以0.1%甲酸乙腈提取30 min,HLB固相萃取柱净化,0.1%甲酸甲醇洗脱时,恩诺沙星、环丙沙星、氧氟沙星、庆大霉素、替米考星、泰乐菌素6种药物质谱信息捕捉准确,色谱分离效果良好,在限定浓度范围内标准曲线回归系数R2均大于0.99,在5、20、50μg/kg 3个加标浓度下回收率为73.3%~114.0%,相对标准偏差(n=6)1.01%~13.01%。6种药物在羊3种可食组织中检出限(LOD)0.05~1.2μg/kg,定量限(LOQ)0.16~3.8μg/kg,均低于现行国家标准中规定的该类药物残留检出限。单因素方差分析结果显示,方法与国家标准检测方法在检测结果上差异不显著(P>0.05)。【结论】建立了羊可食组织中恩诺沙星、环丙沙星、氧氟沙星、庆大霉素、替米考星、泰乐菌素药物残留检测的高效液相色谱串联质谱法,该方法灵敏、准确、高效,可为羊肉及其副产品中抗微生物药物残留监测、筛选和精准识别提供技术支撑。

基于硅烷化三聚氰胺海绵的改良QuEChERS结合UPLC-MS/MS快速测定猪肉中49种抗生素多残留

采用十八烷基三氯硅烷(octadecyltrichlorosilane,OTS)与N-[3-(三甲氧基硅基)丙基]乙二胺(N-[3-(trimethoxysilyl)propyl]ethylenediamine,ATS)改性三聚氰胺海绵(melamine sponge,MeS)制备得到2种新型弹性多孔硅烷化MeS(OTS@MeS和ATS@MeS)净化材料,将其用于改良QuEChERS(quick,easy,cheap,effective,rugged and safe)基质净化过程,通过溶液自发浸润和物理挤压过程即可快速、高效分离干扰基质,结合超高效液相色谱-串联质谱技术建立了同时测定猪肉中49种抗生素多残留的分析方法。样品用1.0%乙酸-乙腈溶液提取并用2.0 g Na_(2)SO_(4)和0.5 g NaCl盐析,离心后取1 mL上清液复配使用OTS@MeS与ATS@MeS净化,随后使用Agilent ZORBAX Eclipse Plus C_(18)色谱柱,以甲醇溶液和含有0.1%甲酸、5 mmol/L乙酸铵的甲醇-水溶液(5∶95,V/V)为流动相进行梯度洗脱,采用电喷雾电离源、正离子多反应监测模式进行定性定量分析。结果表明:49种抗生素的相关系数均大于0.999,基质效应为-13.5%~10.9%,检出限在0.1~10μg/kg之间,定量限在0.3~30.3μg/kg之间;利用该方法在低、中、高3个加标水平下进行回收率实验,回收率范围为65.0%~112.7%,日内和日间相对标准偏差分别为0.3%~11.8%和2.4%~18.4%。该方法样品前处理过程简单、快捷、灵敏度高、准确性好,可用于猪肉中49种抗生素多残留的高效快速检测。

UPLC-MS/MS法同时测定蛋白饮料和液体调味品中36种防腐剂

采用提取和净化相结合的前处理方式,建立并优化了同时测定蛋白饮料、液体调味品中36种防腐剂的超高效液相色谱-串联质谱检测方法。样品采用饱和NaCl溶液(用磷酸调节pH=3)净化和乙腈-甲醇(9∶1,体积比)(含体积分数为0.2%的甲酸)提取。采用C18反向色谱柱进行分离,以甲醇和5 mmol/L乙酸铵为流动相梯度洗脱,采用电喷雾离子源正负离子同时扫描,动态多反应监测模式,基质匹配标准曲线,外标法定量。结果表明,36种防腐剂在1~250 ng/mL线性良好(相关系数≥0.999),方法定量限为0.04~0.2 mg/kg;空白样品不同加标水平下的平均加标回收率为75%~119%;相对标准偏差为0.90%~9.8%。建立的高通量检测方法灵敏、快速,准确度高,操作简便且能有效减少基质干扰、降低检测成本,极大提高多种防腐剂的定性定量检测效率。

UPLC/MS-MS结合QuEChERs检测可食性鸡组织中8-α-羟基妙林残留

为准确测定可食性鸡组织中8-α-羟基妙林的残留量,试验建立了一种高效液相色谱-串联质谱(UPLC/MS-MS)结合QuEChERs的快速检测方法。样品经酸化丙酮提取,Bond Elut QuEChERs柱净化,氢氧化钠碱溶液水解,乙酸乙酯反萃取,高效液相色谱-串联质谱仪测定。结果显示:鸡肉和鸡肝中8-α-羟基妙林在50~200μg/kg浓度范围内,方法具有良好的线性相关性,相关系数≥0.99;在可食性鸡组织中的检出限为10~20μg/kg,定量限为25~50μg/kg;8-α-羟基妙林在50、100和200μg/kg添加水平下平均回收率为65.07%~103.16%,批内、批间变异系数均≤20%。研究提示:建立的方法操作简便、结果准确,满足了兽药残留药物痕量检测方法规定要求,适用于可食性鸡组织中8-α-羟基妙林的残留量测定。

HPLC-MS/MS法快速测定电子烟油中58种合成大麻素

本文旨在建立一套高效液相色谱-串联质谱法(HPLC-MS/MS)快速定性定量检测电子烟油中58种合成大麻素的分析方法。在研究中,选择适宜液相色谱柱,对色谱条件和质谱条件进行优化。样品中的目标物经甲醇萃取后,使用五氟苯基丙基(PFPP)色谱柱(100 mm×2.1 mm×5μm)分离,以乙腈和20 mmol/L乙酸铵+0.1%(体积分数)甲酸缓冲水溶液为流动相进行梯度洗脱,在电喷雾离子源正离子(ESI+)模式下,采用动态多反应监测模式(d-MRM)测定,使用特征离子碎片的峰面积进行外标法定量。结果表明,58种合成大麻素在0.5~50 ng/mL浓度范围内线性关系良好,R2均大于0.99;检出限(LOD,S/N=3)为0.5~1μg/g,定量限(LOQ,S/N=10)均为2μg/g;58种合成大麻素在空白基质中高、中和低三个添加水平下的平均回收率为89.9%~144.0%,相对标准偏差均小于15%;基质效应考察结果表明,78%的合成大麻素呈现为基质增强效应,其他为基质抑制效应;仅5%的合成大麻素表现为强基质效应,其他为中等或弱基质效应;采用基质匹配标准曲线校正,可有效降低基质效应的影响。将该方法应用于本年度内深圳市办案机关提供的电子烟油检测,其中涉及的6种合成大麻素均被准确筛查和定量检测。该方法操作简单,检测覆盖范围广,耗时少,准确度高且检出限低,适用于电子烟油中合成大麻素的快速筛查和定量分析。

HPLC-DAD-ESI-MS/MS结合分子网络技术分析彭祖寿柑果肉的化学成分

以彭祖寿柑(Citrus reticulata Blanco)果肉为材料,采用高效液相色谱-二极管阵列检测器-电喷雾电离质谱(HPLC-DAD-ESI-MS/MS)结合分子网络技术对果肉成分进行分析。采用Thermo Hypersil GOLD C18色谱柱(250 mm×4.6 mm,5μm),以甲醇-0.1%甲酸水为流动相进行梯度洗脱。在正负离子模式下采集质谱数据,并上传至全球天然产物社会分子网络(Global Natural Products Social Molecular Networking,GNPS)创建彭祖寿柑果肉分子网络。根据文献报道及在线数据库mzcloud、pubchem,比对二级质谱碎片信息及紫外吸收特征,排除分子网络假阳性结果,在果肉中共注释出70个成分,包括黄酮类7个、苯丙素类25个、二苯基庚烷类8个、有机酸类11个、脂肪酸类5个、氨基酸类4个、萜类4个、核苷酸类2个、生物碱类1个与其他类3个,其中8个二苯基庚烷成分和2个木脂素成分可能为其潜在的新成分。该研究将液相色谱-质谱联用技术与分子网络技术结合,能够快速、准确、较全面地注释彭祖寿柑果肉成分,为彭祖寿柑果肉进一步的开发与利用提供理论参考。