分散固相萃取/高效液相色谱-离子阱飞行时间高分辨质谱法对纺织品中全氟化合物的快速筛查和确证

采用液相色谱离子阱飞行时间串联质谱(LC-MS-IT-TOF)技术,结合分散固相萃取净化方法,建立了纺织品中全氟化合物快速筛查、定性识别和准确定量的分析方法,构建了14种全氟化合物的多级碎片离子质谱图数据库。样品用甲醇超声提取,提取液经C18吸附剂净化后,以C8色谱柱(150 mm×4.6 mm,3.5μm)分离,5 mmol/L乙酸铵-甲醇为流动相梯度洗脱。考察了吸附剂种类和用量对加标回收率的影响,优化了净化条件。结果显示,14种全氟化合物的检出限为1~10μg/kg,在10,50,100μg/kg 3个加标水平下的平均回收率为49.2%~101.4%,相对标准偏差为1.1%~14.0%。该方法利用精确质量数匹配和自建标准谱库检索进行快速筛查,并使用保留时间和多级特征碎片离子进行定性确证,具有简便、快速、高效、准确等优点,可用于纺织品中全氟化合物的快速筛查和确证。     

HPLC法快速测定熟肉制品中10种食品添加剂

建立了熟肉制品中10种食品添加剂的高效液相色谱分析方法。以乙醇–氨水溶液为提取溶剂,样品经超声波辅助溶剂萃取法萃取后,以Atlantis d C18(250 mm×4.6 mm,5μm)色谱柱为分离柱,20 mmol/L乙酸铵–甲醇为流动相进行分离测定。10种目标化合物的质量浓度在1.0~200.0 mg/L范围内与其峰面积线性关系良好(r0.999),在20.0,500.0,1 000.0 mg/kg的添加水平下,10种目标化合物的平均加标回收率为82%~104%,测定结果的相对标准偏差小于4.0%(n=6),方法的检出限为1.0~5.0 mg/kg(S/N=3)。该方法简单、快速、准确,适合于熟肉制品中食品添加剂的检测。     

高效液相色谱法同时快速检测腌腊肉制品中20种非法添加合成色素

采用亚3μm色谱柱建立了同时快速检测腌腊肉制品中20种非法添加合成色素的高效液相色谱法。样品经石油醚脱脂、乙醇-氨水-水(7∶2∶1)提取、钨酸钠溶液沉淀蛋白,采用Agilent Poroshell 120 EC C18(50 mm×4.6 mm,2.7μm)色谱柱以甲醇-0.02 mol/L乙酸铵(p H 5.5)为流动相梯度洗脱,二极管阵列检测器检测,外标法定量。结果表明,待测物可在17 min内实现完全分离。20种成分的质量浓度在0.15~20.0 mg/L范围内与峰面积呈线性关系,相关系数(r)大于0.998,方法的回收率为70.3%~107.9%,相对标准偏差(RSD)为0.5%~3.1%,检出限为0.1~0.2 mg/kg。与传统5μm色谱柱ACE C18(250 mm×4.6 mm,5μm)相比,本方法节约了62%的分析时间,并能在常规HPLC系统内使用,具有广阔的应用前景。     

二维液相色谱法在线富集测定银杏叶提取物中银杏酸含量

通过2台高压恒流泵、1台紫外检测器、1个手动进样阀、1个高压切换阀和2根色谱柱组建了二维液相色谱系统,建立了在线富集测定银杏叶提取物(EGB)及其制剂中微量银杏酸的方法。第一维色谱柱为富集柱,采用ZORBAX Eclipse Plus-C18(2.1 mm×12.5 mm,5μm),流动相为甲醇-水-三氟乙酸混合液(体积比80∶20∶0.01),流速为0.25 m L/min,可实现300μL样品的富集,有效降低了基体干扰。第二维色谱柱为分析柱,采用ZORBAX Eclipse Plus-C18(2.1 mm×150 mm,5μm),流动相为甲醇-水-三氟乙酸混合液(体积比90∶10∶0.01),流速为0.25 m L/min,可有效分离银杏酸。使用紫外检测器在310 nm处检测,可实现对银杏酸的准确定量。研究了试样的溶剂对富集效果的影响,当试样溶剂与第一维流动相一致时可获得较好的富集效果。该方法对试样中5种银杏酸在0.200~100.0 mg/kg范围内具有良好的线性关系,相关系数均大于0.998,检出限达0.02~0.06 mg/kg。对实际样品测定结果的相对标准偏差均不大于5.0%,在5、10 mg/kg加标水平下,加标回收率为94.0%~101.3%。     

液相色谱-串联质谱法同时测定调制乳制品中5种人工合成甜味剂

建立了调制乳制品中安赛蜜、甜蜜素、糖精钠、阿斯巴甜和纽甜等5种人工合成甜味剂的液相色谱-串联质谱测定方法。对蛋白沉淀剂、色谱柱、流动相组成、超声时间以及质谱参数进行了优化。结果表明,样品经过0.2%乙酸水沉淀蛋白,MGⅢ-C18色谱柱(150 mm×2.0 mm,5μm)分离,甲醇和10 mmol/L乙酸铵水溶液流动相进行梯度洗脱,流速0.3 m L/min,采用液相色谱-串联质谱仪对目标化合物进行测定,基质外标法定量。在10~1020μg/kg范围内,线性相关系数(R2)0.999,3个不同浓度添加水平下平均加标回收率90.0%~109.7%,精密度(RSD)为0.8%~7.6%(n=6)。调制乳制品中安赛蜜、甜蜜素、糖精钠、阿斯巴甜和纽甜的检出限分别为3.0,30,10,5和1.0μg/kg,定量限为10,100,40,20和3.3μg/kg。采用所建立的方法对市面上销售的8种调制乳中5种人工合成甜味剂进行筛查,达到预期效果。方法适用于调制乳制品中人工合成甜味剂的快速分析。     

液相色谱-串联质谱法快速筛查减肥类中药及保健品中非法添加30种化学药

建立了中药及保健品中30种减肥类化学药的高效液相色谱-串联质谱(LC-MS/MS)快速筛查方法。实验优化了前处理方法、色谱分离条件和质谱参数。样品采用甲醇超声萃取,Agilent poroshell 120 ECC18(4.6 mm×100 mm,2.7μm)色谱柱分离,流动相为乙腈-水溶液(含10 mmol/L甲酸铵,0.1%甲酸,10%甲醇),梯度洗脱,流速0.25 m L/min,采用正负离子切换模式的电喷雾质谱检测,多反应选择离子监测(MRM)。该方法能同时筛查30种化学药,覆盖面广,简便、快速、准确可靠,已用于减肥类中药及保健品中非法添加化学药的筛查及检测。     

降压类中成药及保健食品中21种非法添加化学药物的快速检测与确证方法研究

采用亚3μm色谱柱建立了降压类中成药及保健食品中21种非法添加化学药物的高效液相色谱快速检测及液相色谱-质谱联用确证方法。样品经甲醇-乙腈(体积比5∶5)超声提取,采用Agilent Poroshell 120Phenyl-Hexyl(100 mm×4.6 mm,2.7μm)色谱柱,以甲醇-乙腈(体积比2∶1)-甲酸水溶液(p H 2.5±0.1)为流动相,梯度洗脱,二极管阵列检测器检测,外标法定量,液质联用法进一步定性确证。结果表明,21种成分在17 min内完成分离,方法检出限为0.03~0.50 mg/g,定量下限为0.09~1.50 mg/g,平均回收率为82.0%~109.0%。采用上述方法对107批从互联网收集的样品进行检测,阳性检出率为42.1%。该方法快速、准确,适用于降压类中成药及保健食品中非法添加药物的快速检测。     

高效液相色谱法测定食品中的爱德万甜

建立了食品中爱德万甜检测的高效液相色谱分析方法。样品经甲醇-水(50∶50,体积比)溶液提取10 min,离心后取上清液进行分析。采用Agilent TC-C18(2)(250 mm×4.6 mm,5μm)色谱柱分离,以甲醇-0.02 mol/L乙酸铵(53∶47,体积比)等度洗脱,在280 nm波长下检测,外标法定量。结果表明,目标化合物在1.0~200.0 mg/L范围内呈良好的线性,相关系数(r)为0.999 8,饮料、酸奶和蛋糕的检出限(S/N≥3)分别为0.6、1.5、9.0 mg/kg。方法的回收率为90.9%~103.4%,相对标准偏差(RSD,n=5)为1.1%~4.5%。该方法的色谱分离效果好,干扰少,准确度高,可用于实际样品中爱德万甜的测定。     

QuEChERS前处理结合HPLC-Q-TOF/MS非靶向快速筛查凉茶中的非法添加物

建立了QuEChERS法净化,结合高效液相色谱-四极杆-飞行时间质谱法(HPLC-Q-TOF/MS)非靶向快速筛查凉茶中未知非法添加物的方法。凉茶中的非法添加物经0.5%乙酸-乙腈提取,氨丙基粉净化,C_(18)色谱柱(100 mm×2.1 mm,1.7μm)分离,HPLC-Q-TOF/MS测定。利用8种质控化合物建立的筛查方法检出限为2.0～10μg/kg,定量下限为5.0～25μg/kg。3个不同加标水平的平均回收率为75.8%～95.6%,RSD(n=6)为2.1%～6.5%。在该定量方法基础上利用目标化合物特征离子的精确质量数、同位素匹配、二级碎片信息进行数据库匹配,无标准品情况下非靶向筛查非法添加物。结果表明,该方法无需标准品即可快速筛查凉茶中的非法添加西药,能应用于凉茶样品的实际筛查。应用该方法对50份散装凉茶样品进行测定,其中10份凉茶样品检出对乙酰氨基酚,含量为4.14～2 188 mg/kg。该方法快速、准确、分析通量高,可为凉茶中非法添加物的快速筛查和质量控制提供重要依据。     

亚3微米色谱分离-串联质谱法同时筛查中药材中非法添加的30种人工色素EI北大核心CSCD

建立了中药材中30种人工色素的高效液相色谱-串联质谱(LC-MS/MS)筛查测定方法。试验优化了前处理方法,分离条件和质谱参数。样品采用甲醇超声,提取液经亚3μm填料色谱柱Agilent Poroshell 120 EC-C18(3.0 mm×100 mm,2.7μm)分离,乙腈-水(10 mmol/L乙酸铵)为流动相梯度洗脱,流速0.30 m L/min,采用电喷雾离子源,正、负离子切换采集模式多反应选择离子监测(MRM)。30种人工色素及其中的3对同分异构体得到了理想分离和定性定量。本法简便、快速、准确可靠,已经应用于中药材中非法添加的人工色素的筛查及检测。     

高效液相色谱-质谱法测定降血压保健食品中的8种违禁添加成分

建立了降血压保健食品中8种违禁添加成分的高效液相色谱-电喷雾质谱联用(HPLC-ESI/MS)测定方法。采用Spherigel C18反相色谱柱,以0.005 mol/L甲酸铵缓冲盐(pH 3.0)-乙腈-甲醇为流动相,梯度洗脱,质谱定性定量。在该条件下,以西力士为内标,ESI+模式下的最低检出限(LOD)为2.5 μg/L;ESI-模式下的最低检出限为50 μg/L;回收率为63.3%～107.4%。该方法的样品处理简单,结果准确,重现性好,选择性及灵敏度高,适用范围广,可用于降压类药物及保健食品中违禁成分的检测。     

高效液相色谱-四极杆/静电场轨道阱高分辨率质谱法快速筛查中成药和保健食品中非法添加的42种化学药物

建立了高效液相色谱-四极杆/静电场轨道阱高分辨率质谱法快速筛查中成药和保健品中非法添加化学药物的方法。中成药和保健品样品,经提取溶剂提取,以12000 r/min离心后进行质谱分析。采用Phe-nomenex C18色谱柱(100 mm×4.6 mm,2.6μm)进行分离,以乙腈和0.1%甲酸溶液作为流动相,进行梯度洗脱。质谱采用正离子和负离子同时扫描,Fullms-dd-MS2模式进行分析。在所建立的色谱条件下,42种化学药物能够得到较好分离,在线性范围内线性关系良好,相关系数均大于0.99。通过加标验证,在20,50和100 ng/g加标水平下,所有分析物的平均回收率为69.3%~105.2%,相对标准偏差(RSD)小于8.9%。运用本方法对31种保健品和中成药进行快速筛查,发现其中3种添加了盐酸二甲双胍,1种添加了西地那非,1种添加了羟基莫豪西地那非。本方法样品处理过程简单,分析时间短,准确可靠,灵敏度高。适用于中成药和保健品中非法添加化学药品的定性与定量检测,可用于非法添加药物的筛查。     

Reverse-phase liquid chromatographic determination of α-lipoic acid in dietary supplements using a boron-doped diamond electrode

A fast liquid chromatographic separation, coupled with sensitive and straightforward detection using a boron-doped diamond (BDD) electrode, was developed and validated for the determination of α-lipoic acid in dietary supplement samples. The analysis was carried out using a reversed phase C18 (150 mm × 4.6 mm, 5 μm) column with a mobile phase consisting of a 1:1 (v/v) ratio of 0.05 M phosphate solution (pH 2.5):acetonitrile, at a flow rate of 1.0 mL/min. The detection potential obtained from hydrodynamic voltammetry was 1.05 V vs. Ag/AgCl. Under optimized conditions, the chromatographic separation was performed in less than 5 min, a good linear relationship was obtained between the current and the α-lipoic concentration within the range of 0.01-60 μg/mL (correlation coefficient of 0.9971), and a detection limit of 3.0 ng/mL was determined. Furthermore, this method was successfully applied to determine α-lipoic acid concentrations in selected commercial dietary supplement samples. The recovery of α-lipoic acid in spiked samples at 0.5, 5.0 and 30 μg/mL ranged from 94.4% to 103.6% with a relative standard deviation (RSD) of between 1.2% and 3.7%. In real samples, this developed methodology produced results that were highly correlated with the standard HPLC-UV approach. Therefore, the present method can be used for fast, selective and sensitive quantification of α-lipoic acid in dietary supplements.     

Determination of ephedrine alkaloids in dietary supplements and botanicals by liquid chromatography/tandem mass spectrometry: collaborative study

An interlaboratory study was conducted to evaluate the accuracy and precision of a method for ephedrine-type alkaloids [i.e., norephedrine (NE), norpseudoephedrine (NPE), ephedrine (E), pseudoephedrine (PE), methylephedrine (ME), and methylpseudoephedrine (MPE)] in dietary supplements and botanicals. The amount of ephedrine-type alkaloids present was determined using liquid chromatography with tandem mass selective detection. The samples were diluted to reflect a concentration of 0.0200 to 1.00 microg/mL for each alkaloid. An internal standard was added and the alkaloids were separated using a 5 microm phenyl LC column with an ammonium acetate, glacial acetic acid, acetonitrile, and water mobile phase. Eight blind duplicates of dietary supplements or botanicals were analyzed by 10 collaborators. Included was a negative control, ephedra nevadensis, and negative controls fortified at 2 different levels with each of the 6 ephedrine-type alkaloids. The spike levels were approximately 100 and 1000 microg/g for NE, 100 and 600 microg/g for NPE, 6500 and 65000 microg/g for E, 1000 and 10 000 microg/g for PE, 300 and 3000 microg/g for ME, and 100 and 1000 microg/g for MPE. On the basis of the accuracy and precision results for this interlaboratory study, it is recommended that this method be adopted Official First Action for the determination of 6 different individual ephedrine-type alkaloids in dietary supplements and botanicals.     

LC of sulfonamide residues in poultry muscle and eggs extracts using fluorescence pre‐column derivatization and monolithic silica column

A new, rapid, sensitive and selective HPLC method with fluorescence detection is described for the simultaneous determination of 12 sulfonamides, in the presence of putrescine as internal standard, after pre‐column derivatization with fluorescamine. The drugs were separated on a Chromolith Performance RP‐18 column (100×4.6 mm), using a gradient elution with a binary mobile phase of methanol/0.05 M acetate buffer (pH 3.4). Linearity of derivatization was obtained for concentrations from 3.0 to 300 μg/L in standard solutions. The whole procedure was evaluated and fully validated, according to the European Union Decision 2002/657/EC, for the determination of sulfonamides in turkey muscle and hen eggs following SPE. The LODs varied from 2 to 17 μg/kg in turkey and 2 to 15 μg/kg in egg samples. The average recoveries ranged between 96.9–108.6% in turkey muscle and 96.0–108.4% in egg samples, respectively.     

超高效液相色谱-串联质谱法同时测定保健食品中丙磺舒、别嘌醇和苯溴马隆

建立了基于QuEChERS前处理的超高效液相色谱-串联质谱（UPLC-MS/MS）同时测定抗痛风类保健食品中别嘌醇、丙磺舒和苯溴马隆3种药物的检测方法。样品经含0.1%（v/v）氨水的乙腈溶液超声提取后，采用乙二胺-N-丙基硅烷（PSA）和十八烷基键合硅胶（C₁₈）吸附剂净化，用C₁₈色谱柱进行分离，0.1%（v/v）甲酸水溶液和甲醇为流动相进行梯度洗脱，采用电喷雾电离源、正负离子切换模式和多反应监测（MRM）模式检测。结果表明，别嘌醇、丙磺舒和苯溴马隆的检出限分别为5、25和25 μg/kg，定量限分别为17、80和80 μg/kg。抗痛风类保健食品中3种药物的平均加标回收率为76.8%~116.6%，相对标准偏差（RSD）为2.7%~14.6%。应用该法对68批次保健食品进行分析，其中1批次样品检出别嘌醇药物。该法操作简单，灵敏度高，可用于抗痛风类保健食品中别嘌醇、丙磺舒和苯溴马隆的测定。     

高效液相色谱-串联质谱法同时测定保健品及中成药中非法添加的17种壮阳类化学药

建立了高效液相色谱-串联质谱(LC-MS/MS)同时测定补肾壮阳类保健品及中成药中非法添加的17种壮阳化学药的定性、定量分析方法。实验优化了前处理方法,并针对几对同分异构体成分优化了分离条件和质谱参数。样品经甲醇超声萃取,提取液经Aglient Extend C18色谱柱(100 mm×2.1 mm, 3.5 μm)分离,流动相为乙腈和水(含有10 mmol/L乙酸铵),梯度洗脱,流速为0.25 mL/min,以电喷雾离子源正离子多反应监测(MRM)模式进行MS/MS检测。该方法能很好地分离并定量17种壮阳类化学药以及其中的3组同分异构体。该方法简便、快速、准确可靠,已经应用于补肾壮阳类保健品及中成药中非法添加壮阳类化学药的筛查及检测。统计近3年检测的样品,发现补肾壮阳类产品中非法添加成分检出率高,需引起相关监管部门的重视。     

A rapid and reliable UPLC-MS/MS method for the identification and quantification of fourteen synthetic anti-diabetic drugs in adulterated Chinese proprietary medicines and dietary supplements

An ultraperformance liquid chromatography–tandem mass spectrometry (UPLC‐MS/MS) method was developed for simultaneous qualitative and quantitative analysis of 14 synthetic anti‐diabetic drugs in adulterated Chinese proprietary medicines (CPMs) and dietary supplements. The samples were prepared by ultrasonic extraction with methanol and separated on a C₁₈ column with mobile phase consisting of acetonitrile and water (both containing 0.10% formic acid). Gradient elution was applied with a flow rate of 0.20 mL/min. Two transitions from protonated molecules were monitored for each synthetic anti‐diabetic drug in positive mode of electrospray ionization (ESI). The two transitions, the peak area ratio of the two transitions and the retention time were used for identification. The more intensive transition was used for quantification. The analysis time was 6 min per sample. Satisfactory linear relationships were estimated between the peak area and the concentration with correlation coefficients higher than 0.995. The limit of detection ranged from 0.03 to 5.45 ng/mL. The relative standard deviation of intra‐day precision was below 7.6%, the RSD of inter‐day precision was below 15% and the relative error of accuracy was between –10 and 7.8%. The proposed method is rapid, selective, reliable and was successfully applied to the analysis of 30 real samples of 22 CPMs and eight dietary supplements from the local market in China. Copyright © 2010 John Wiley & Sons, Ltd.     

Simultaneous Determination of Atenolol,Rosuvastatin, Spironolactone,Glibenclamide and Naproxen Sodium in Pharmaceutical Formulations and Human Plasma by RP‐HPLC

A rapid and sensitive high‐performance liquid chromatographic method was developed and validated for the simultaneous determination and quantification of atenolol, rosuvastatin, spirnolactone, glibenclamide and naproxen sodium in bulk drugs, pharmaceutical formulations and in human plasma in the presence of internal standard (flurbiprofen). Chromatograms were developed with methanol and water (80:20, v/v) solvent system on a Purospher start, C₁₈ (5 μm, 250 × 4.6 mm) column and pH was adjusted to 3.40 with ortho‐phosphoric acid. Mobile phase was pumped with a flow rate of 0.90 mL/minute with 235 nm UV detection. Standard curves were linear over the concentration range 0.25‐30 μg/mL^?1. The coefficients of variation (C.V.%), were < 3% and LOD and LOQ were <0.0154 & 0.06 for inter‐ and intra‐day, respectively. The method was applied to drug interaction studies of atenolol with rosuvastatin, spironolactone, glibenclamide and naproxen to illustrate the scope and application of the methods to manage four different therapeutic classes of drugs, as they are co‐administered.     

Determination of key flavonoid aglycones by means of nano‐LC for the analysis of dietary supplements and food matrices

A method for the analysis of flavonoids (myricetin, quercetin, naringenin, hesperitin, and kaempferol), with interesting bioactivity, has been developed and validated utilizing nano‐LC technique. In order to find optimal conditions, capillary columns (75 μm id × 10 cm) packed with different types of stationary phases, Kinetex® C18 core–shell (2.6 μm particle size), Hydride‐based RP‐C18 (sub‐2 μm particle size), and LiChrospher® 100 RP‐18 endcapped (5 μm particle size) were evaluated. The method was validated using Hydride‐based RP‐C18 stationary phase, with sub‐2 μm particle size. A good chromatographic performance, expressed in terms of repeatability (RSD, in the range 1.63–4.68% for peak area), column‐to‐column reproducibility (RSD not higher than 8.01% for peak area), good linearity and sensitivity was obtained. In particular limit of detection values between 0.07 and 0.31 μg/mL were achieved with on column focusing technique. The method was applied to the determination of studied flavonoids in dietary supplements as well as in food matrices. The amount of quercetin found in the first analyzed dietary supplement, was in agreement to the labeled content. In the other samples, where the content of flavonoids was not labeled, most of the studied flavonoids were determined in amounts somewhere comparable to those reported in literature.