超高压液相色谱-串联四极杆质谱法对牛奶中24种磺胺类药物残留的检测

利用超高压液相色谱-电喷雾串联四极杆质谱（UPLC-MS/MS）联用技术,建立了一种能在10 min内快速分离和测定牛奶中24种磺胺类药物残留的方法。样品经匀浆、超声、乙腈重复提取、氮吹浓缩,流动相溶解,饱和正己烷脱脂。采用ACQUITY UPLCTMBEHC18柱（100 mm×2.1 mm i.d.,1.7μm）,以乙腈-0.2%乙酸水溶液（体积比1∶9）为流动相,梯度洗脱,目标分析物使用超高压液相色谱-电喷雾串联质谱进行测定;以保留时间和离子对进行定性和定量,在ESI（＋）和MRM监测模式下进行样品分析。该方法检出限（LOD）为0.04～1.35μg/kg;在1～200μg/L范围内线性关系良好,回收率为61%～117%,相对标准偏差为2.92%～18.98%。该法样品前处理简单、分析速度快、回收率和灵敏度高、检出限低,可以满足各国对牛奶中24种磺胺类药物的检测要求。     

Comparison of LC and UPLC Coupled to MS–MS for the Determination of Sulfonamides in Egg and Honey

Determination of ten sulfonamides (SAs) in egg and honey has been compared using column liquid chromatography (LC) and ultra-performance liquid chromatography (UPLC) coupled to tandem mass spectrometry (MS–MS). A liquid–liquid extraction with acetonitrile followed by solid-phase extraction on a Strata-X cartridge was developed for sample preparation. The analytical performance of both methods was compared applying the alternative matrix-comprehensive in-house validation approach using specially designed software InterVal?. Using UPLC the separation time was shortened about 30% reducing the run time by 8 min and a better resolution was achieved compared to LC. Due to higher peak efficiency achieved with UPLC, the decision limit values obtained by both techniques were almost equal (6.61–9.43 μg kg^?1 and 7.25–11.9 μg kg^?1 for UPLC and LC, respectively), despite the fact that in UPLC twice lower sample volumes were injected. Satisfactory and comparable recoveries (80–110%) were obtained by UPLC and LC for all the SAs, except for sulfacetamide by LC and sulfabenzamide by both methods. For a majority of the spiked compounds, UPLC gave significantly better precision.     

Monitoring of sulfonamide antibacterial residues in milk and egg by polymer monolith microextraction coupled to hydrophilic interaction chromatography/mass spectrometry

A simple, rapid, and sensitive method for the determination of traces of thirteen sulfonamide antibacterials in milk and eggs is presented. This method is based on the combination of polymer monolith microextraction (PMME) technique with hydrophilic interaction chromatography/mass spectrometry (HILIC/MS). The extraction was performed with a poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column while the subsequent separation was carried out on a Luna NH₂ column by HILIC. To obtain optimum results, several parameters relating to HILIC and PMME were investigated. After optimization, acetonitrile (contain 0.05% formic acid, v/v) was used as the elution solution, which was well compatible with the mobile phase in HILIC. Good linearities were obtained for thirteen SAs with the correlation coefficients (R²) above 0.997. The limits of detection (S/N = 3) of the method were found to be 0.4–5.7 ng mL⁻¹ of SAs in whole milk and 0.9–9.8 ng g⁻¹ of SAs in eggs. The recoveries of thirteen SAs in two matrices ranged from 80.4 to 119.8%, with relative standard deviations less than 11.8%.     

A novelty strategy for the fast analysis of sulfonamide antibiotics in fish tissue using magnetic separation with high‐performance liquid chromatography–tandem mass spectrometry

A simple, fast and low‐cost extraction method with high‐performance liquid chromatography–tandem mass spectrometry (HPLC‐MS/MS) determination was developed on sulfonamide antibiotics (SAs) in fish tissue. Magnetic separation was first introduced into the rapid sample preparation procedure combined with acetonitrile extraction for the analysis of SAs. Partitioning was rapidly achieved between acetonitrile solution and solid matrix by applying an external magnetic field. Acetonitrile solution was collected and concentrated under a nitrogen stream. The residue was redissolved with 1‰ formic acid aqueous solution and defatted with n‐hexane before analysis. The recoveries of SAs were in the range of 74.87–104.74%, with relative standard deviations <13%. The limits of quantification and the limits of detection for SAs ranged from 5.0 to 25.0 μg ^kg?1 and from 2.5 to 10.0 μg ^kg?1, respectively. The presented extraction method proved to be a rapid method which only took 20 min for one sample preparation procedure. Copyright © 2016 John Wiley & Sons, Ltd.     

Rapid and Sensitive UPLC-MS–MS for the Determination of Trimetazidine in Human Plasma

A specific and sensitive UPLC-MS–MS was developed for the determination of trimetazidine in human plasma. The sample preparation was based on a single-step liquid–liquid extraction with acetic ether. The chromatographic separation was on a C₁₈ analytical column (50 mm × 2.1 mm, 1.7 μm) with acetonitrile and 10 mM ammonium acetate (30:70, v/v) as the mobile phase, and a triple-quadrupole mass spectrometer equipped with an electrospray ionization source (ESI) applied for detection. The method was linear over the concentration ranges of 0.25–100.00 ng mL⁻¹ for trimetazidine, and the lower limit of quantification (LLOQ) was 0.25 ng mL⁻¹. The intra- and inter-day relative standard deviation (RSD) were less than 15% and the relative error (RE) were all within 15%. Finally, this method has been successfully applied to analyze plasma samples from a bioequivalence study with 18 volunteers.

     

Rapid and Sensitive UPLC-MS–MS for the Determination of Trimetazidine in Human Plasma

A specific and sensitive UPLC-MS–MS was developed for the determination of trimetazidine in human plasma. The sample preparation was based on a single-step liquid–liquid extraction with acetic ether. The chromatographic separation was on a C₁₈ analytical column (50 mm × 2.1 mm, 1.7 μm) with acetonitrile and 10 mM ammonium acetate (30:70, v/v) as the mobile phase, and a triple-quadrupole mass spectrometer equipped with an electrospray ionization source (ESI) applied for detection. The method was linear over the concentration ranges of 0.25–100.00 ng mL^?1 for trimetazidine, and the lower limit of quantification (LLOQ) was 0.25 ng mL^?1. The intra- and inter-day relative standard deviation (RSD) were less than 15% and the relative error (RE) were all within 15%. Finally, this method has been successfully applied to analyze plasma samples from a bioequivalence study with 18 volunteers.     

Analysis of toldimfos in porcine muscle and bovine milk using liquid chromatography–triple quadrupole mass spectrometry

An analytical method was developed for the detection of toldimfos sodium residues in porcine muscle and bovine milk using liquid chromatography–triple quadrupole tandem mass spectrometry (LC–MS/MS) analysis. The drug was extracted from muscle and milk using 10 mm ammonium formate in acetonitrile and then purified using n ‐hexane. The drug was well separated on a Luna C₁₈ column using a mixture of 10 mm ammonium formate in ultrapure water (A) and acetonitrile (B) as the mobile phase. Good linearity was achieved over the tested concentration range (0.005–0.03 mg/kg) in matrix‐matched standard calibration. The determination coefficients (R ² ) were 0.9942 and 0.9898 for muscle and milk, respectively. Fortified porcine muscle and bovine milk contained concentrations equivalent to and twice the limit of quantification (0.005 mg/kg) yielded recoveries in the range of 75.58–89.74% and relative standard deviations of ≤8.87%. Samples collected from large markets located in Seoul, Republic of Korea, tested negative for toldimfos sodium residue. In conclusion, ammonium formate in acetonitrile can effectively extract toldimfos sodium from porcine muscle and bovine milk without solid‐phase extraction, which is usually required for cleanup before analysis. This method can be applied for the routine analysis of toldimfos in foods of animal origins.     

Simultaneous determination of 24 sulfonamide residues in meat by ultra-performance liquid chromatography tandem mass spectrometry

The present study used the liquid extraction pretreatment method and developed an ultra-performance liquid chromatography triple quadrupole tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of 24 kinds of sulfonamide residues in meat. The meat samples were homogenized, extracted and deproteinized by acetonitrile, defatted by n-hexane, and further liquid-liquid extracted by ethyl acetate. All of 24 sulfonamide residues were simultaneously separated and determined by UPLC-MS/MS within 15 min. The sulfonamide residues were monitored via the ESI(+) ionization method and quantified by six-channel multiple reaction monitoring (MRM). The calibrations were performed in sample matrixes by the isotope dilution method and the interference effect of sample matrixes on the ionization was effectively eliminated. Good linear relationship (R(2)=0.991-0.999) was observed within the concentration range of 0.2-50 microg/kg. Satisfied recoveries (67.8-113.9%) of all the sulfonamides were demonstrated in different standard-spiked levels except sulfanitran (SNT). The analytical category, separation speed, selectivity, sensitivity and repeatability of sulfonamides using UPLC-MS/MS were significantly improved compared to other analytical methods. Quantitative results of 240 meat samples demonstrated that the present method has a convenient operation and good practicability, which can be applied to the quantitative analysis of a large number of samples.     

Validation Method for the Determination of Sulfonamide Residues in Bovine Milk by HPLC

A simple, rapid, sensitive and reliable high-performance liquid chromatographic method for the simultaneous determination of eight sulfonamides (SAs) in bovine milk was developed (sulfadiazine, sulfathiazole, sulfamethazine, sulfamethoxypyridazine, sufamonomethoxine, sulfamethoxazole, sulfadimethoxine and sulfaquinoxaline) in bovine milk was developed. Samples were prepared by extraction with ethyl acetate and cleaning-up with an anion solid-phase extraction (SPE) column. Analytical separation was performed on an Inertsil ODS-3 column with photodiode-array detection at 270 nm under gradient condition. The whole procedure was evaluated according to the European Commission Decision 2002/657/EC. Specificity, decision limit (CCα), detection capacity (CCβ), trueness and precision were determined during the validation process. It was found that the analytes were isolated from spiked samples with good recoveries between 70.5 and 89.0%. The used analytical conditions allow to successively separating all the tested sulfonamides with good limit of detection between 0.8 and 1.5 μg L⁻¹.     

Simultaneous Determination of Ebastine and Its Active Metabolite (Carebastine) in Human Plasma Using LC–MS–MS

A sensitive and rapid LC–MS–MS method was developed for the simultaneous determination of ebastine and carebastine in human plasma. Solid-phase extraction was used to isolate the compounds from the biological matrix followed by separation on a Symmetry C18 column under isocratic conditions. The mobile phase was 10 mM ammonium formate in water/acetonitrile (40:60, v/v). Detection was carried out using a triple-quadrupole mass spectrometer in positive electrospray ionization and multiple reaction monitoring mode. The method was fully validated over the concentration range of 0.1–10 ng mL^?1 for ebastine and 0.2–200 ng mL^?1 for carebastine in human plasma, respectively. The lower limit of quantification (LLOQ) was 0.1 ng mL^?1 for ebastine and 0.2 ng mL^?1 for carebastine. For ebastine and carebastine inter- and intra-day precision (CV%) and accuracy values were all within ±15% and 85–115%, respectively. The extraction recovery was on average 60.0% for ebastine and 60.3% for carebastine.     

Applications of a Novel Sample Preparation Method for the Determination of Sulfonamides in Edible Meat by CZE

A simple, rapid and reliable method based on SPE clean-up and CZE separation was validated for the trace determination of sulfonamides (SAs) in meat. Acetonitrile was used for the extraction of SAs (sulfamethoxazole, sulfamethazine, sulfamerazine, and sulfadimethoxine) from the samples and 1-propanol was used for the denaturing of the proteins present in the sample matrix. SPE procedure was employed for the clean-up and pre-concentration of SAs prior to CZE analysis. Complete separation was achieved by using 45 mmol L^?1 phosphate buffer (pH = 6.3) at an applied voltage of 20 kV. Overall obtained recoveries were from 83.3 to 94.5% for the SAs. The detection limit of each sulfonamide ranges from 4 to 6 μg kg^?1. The presented one step SPE clean-up method is highly applicable for the determination of the SAs at a residue level below the maximum residue limit.     

超高效液相色谱-串联四极杆质谱法分析婴儿乳粉中的唾液酸

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)测定婴儿配方乳粉中唾液酸含量的分析方法。利用酸水解方法释放出婴儿配方乳中的唾液酸,经HLB反相色谱固相萃取柱净化,采用BEH HILIC色谱柱分离,以0.1%甲酸水溶液和100%乙腈溶液作为流动相进行梯度洗脱,流速为0.25 mL/min,进样体积5 μL,柱温30℃,电喷雾质谱检测,正离子多反应监测模式进行定性和定量分析。结果表明:唾液酸在0.05~5.0 mg/L范围内与唾液酸峰面积的线性关系良好(R²=0.9989);以0.1、0.5、2.5和5.0 mg/L 4个添加水平进行添加回收试验,唾液酸的平均回收率为84.3%~98.9%,相对标准偏差为4.9%~8.2%;唾液酸的检出限为0.01 mg/L。该方法简单、快速、重复性好、灵敏度高,可广泛用于奶粉、牛奶及母乳中唾液酸含量的分析测定。     

Validation of QuEChERS-based UPLC-MS/MS method for determination of quinoid niclosamide (LDS) residue in water,soil and rice samples

A sensitive, rapid and easy analytical method was validated for the determination of quinoid niclosamide (LDS) molluscicide in water, rice and soil using a QuEChERS extraction procedure and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) detection. The LDS was extracted by using acetonitrile and then cleaned up by using dispersive solid-phase extraction with florisil and C18 sorbents. The determination of the target compound was achieved in less than 3 min using an electrospray ionisation source in negative mode. The overall average recoveries for this method in water, rice and soil matrix at three fortified levels ranged from 82.54 to 99.9%, with relative standard deviations in the range of 1.51 to 4.86% (n = 5). The calculated limits of detection were lower than 0.1 µg kg^?1 and quantification was 5 µg kg^?1; these values were much lower than the maximum residue levels established by the Australian standard (0.01 mg kg^?1). The results of the method validation confirmed that this proposed method is convenient and reliable for the determination of LDS molluscicide in water, rice and soil samples.     

Determination of Danofloxacin and Marbofloxacin in Milk Samples by Micellar Liquid Chromatography with Fluorescence Detection

Abstract

A simple, rapid, and sensitive analytical method has been developed for the determination of two fluoroquinolones, danofloxacin and marbofloxacin, in bovine milk samples. Separation and quantification were performed by micellar liquid chromatography with fluorescence detection (MLC?FD), using sodium dodecyl sulfate (SDS) as a surfactant. The influence of the principal factors, namely, the micelle concentration, the amount of organic modifier, tail‐reducing agents, the pH, and the temperature were studied. The suitable condition was found to be 75 mM SDS?10 mM phosphate buffer–18 mM tetrabutylammonium bromide/3% (v/v) 1‐propanol at pH 3.0 for the separation of marbofloxacin, danofloxacin, and tosufloxacin (internal standard) in about 20 min. The linear concentration range of application was 1.8–30.0 ng · mL^?1 for danofloxacin and 16–120 ng · mL^?1 for marbofloxacin, and the relative standard deviation ranged between 4.9 and 2.7%. The limit of detection found for danofloxacin was 0.5 ng · mL^?1 and 5 ng · mL^?1 for marbofloxacin. These values were lower than the maximum residue limits (MRLs) established by the European Union for these compounds in bovine milk. It was applied to check the eventual existence of these compounds above these limits on commercial milk samples. The validation method was completed with spiked milk samples. Recovery levels obtained were 90.3–108.2%.     

Sensitive Analysis of Wilforidine in Human Plasma by LC-APCI-MS/MS

Wilforidine is a potentially efficient medicine to cure autoimmune diseases. In this paper, a sensitive and selective liquid chromatographic method coupled with atmospheric -pressure chemical ionization mass spectrometry (LC–APCI–MS/MS) has been developed for quantification of wilforidine in human plasma. Samples were deproteinized with acetonitrile and cleaned by solid-phase extraction. The chromatographic separation was performed on an analytical RRHD C₁₈ column (50 × 2.1 mm) using ammonium acetate solution (10.0 mmol L⁻¹)/acetonitrile (30/70, v/v) as the mobile phase at a flow rate of 0.7 mL min⁻¹. Detection was carried out by the positive multiple reaction monitoring mode with transitions of m/z 780 → 684 for wilforidine, and 646 → 586 for aconitine (internal standard), respectively. The calibration curve was linear (r = 0.9991) in the concentration range of 0.5–100.0 μg L⁻¹ with a lower limit of quantification of 0.5 μg L⁻¹ in plasma. Intra- and inter-day relative standard deviations were less than 6.8 and 13.1 %, respectively, and the recoveries were between 88.0 and 96.0 %. This accurate and highly specific assay provides a useful method for evaluating the pharmacokinetics of wilforidine in human plasma.

     

Sensitive Analysis of Wilforidine in Human Plasma by LC-APCI-MS/MS

Wilforidine is a potentially efficient medicine to cure autoimmune diseases. In this paper, a sensitive and selective liquid chromatographic method coupled with atmospheric -pressure chemical ionization mass spectrometry (LC–APCI–MS/MS) has been developed for quantification of wilforidine in human plasma. Samples were deproteinized with acetonitrile and cleaned by solid-phase extraction. The chromatographic separation was performed on an analytical RRHD C₁₈ column (50 × 2.1 mm) using ammonium acetate solution (10.0 mmol L^?1)/acetonitrile (30/70, v/v) as the mobile phase at a flow rate of 0.7 mL min^?1. Detection was carried out by the positive multiple reaction monitoring mode with transitions of m/z 780 → 684 for wilforidine, and 646 → 586 for aconitine (internal standard), respectively. The calibration curve was linear (r = 0.9991) in the concentration range of 0.5–100.0 μg L^?1 with a lower limit of quantification of 0.5 μg L^?1 in plasma. Intra- and inter-day relative standard deviations were less than 6.8 and 13.1 %, respectively, and the recoveries were between 88.0 and 96.0 %. This accurate and highly specific assay provides a useful method for evaluating the pharmacokinetics of wilforidine in human plasma.     

LC Method for Determination and Pharmacokinetic Study of 5,6,7,8,3′,4′-Hexamethoxy-3-sulfonyl Flavone in Rat Plasma

A sensitive, specific and rapid high-performance liquid chromatography method was developed for determination of 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone in rat plasma. A simple methanol-induced protein precipitation was applied to extract 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone and Picroside II (the internal standard) from rat plasma. Chromatographic separation was achieved on a Hypersil ODS₂ analytical column (200 mm × 4.6 mm, 5 μm) with acetonitrile–0.04% triethylamine solution (adjusted to pH 5.8 using phosphoric acid) (24:76, v/v) as mobile phase. The calibration curves were linear over the range of 0.2–40 μg mL⁻¹. Absolute recoveries of 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone were 82.7–95.9% from rat plasma. The intra- and inter-day relative standard deviation precisions were less than 5 and 9%, respectively. The method was successfully applied to the pharmacokinetic study of 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone in rats after intravenous administration.

     

LC Method for Determination and Pharmacokinetic Study of 5,6,7,8,3′,4′-Hexamethoxy-3-sulfonyl Flavone in Rat Plasma

A sensitive, specific and rapid high-performance liquid chromatography method was developed for determination of 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone in rat plasma. A simple methanol-induced protein precipitation was applied to extract 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone and Picroside II (the internal standard) from rat plasma. Chromatographic separation was achieved on a Hypersil ODS₂ analytical column (200 mm × 4.6 mm, 5 μm) with acetonitrile–0.04% triethylamine solution (adjusted to pH 5.8 using phosphoric acid) (24:76, v/v) as mobile phase. The calibration curves were linear over the range of 0.2–40 μg mL^?1. Absolute recoveries of 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone were 82.7–95.9% from rat plasma. The intra- and inter-day relative standard deviation precisions were less than 5 and 9%, respectively. The method was successfully applied to the pharmacokinetic study of 5,6,7,8,3′,4′-hexamethoxy-3-sulfonyl flavone in rats after intravenous administration.     

Simple Multiresidue Determination of Fluoroquinolones in Bovine Milk by Liquid Chromatography with Fluorescence Detection

Abstract

A simple and sensitive method for the simultaneous determination of five fluoroquinolones (ciprofloxacin, enrofloxacin, danofloxacin, difloxacin, and sarafloxacin) in bovine milk was developed. Protein precipitation from milk samples was achieved by the addition of acetonitrile and o‐phosphoric acid. Acetonitrile was removed with dichloromethane, leaving the fluoroquinolones in the acid aqueous extract. The aqueous extract was analyzed by liquid chromatography with fluorescence detection (LC–FD). The mobile phase was composed of acetonitrile and 10 mM citrate buffer solution of pH 4.5, with an initial composition of acetonitrile‐water (12∶88, v/v) and using linear gradient elution. Norfloxacin was used as an internal standard. The limits of detection found ranged from 1 to 6 ng · mL^?1 and were below the maximum residue limits (MRLs) established by the European Union. The proposed method was applied to the determination of these compounds in different bovine milk samples. Method validation was carried out by a recovery assay.     

多壁碳纳米管固相萃取净化-高效液相色谱法测定猪肉和鸡肉中的磺胺多残留

建立了多壁碳纳米管为吸附剂的固相萃取净化-高效液相色谱-紫外检测测定猪肉和鸡肉中多种磺胺类药物多残留的方法。样品采用乙腈提取,多壁碳纳米管固相萃取净化,NaH₂PO₄缓冲溶液（pH 5.5~6.0）溶解上样,5%（v/v）丙酮-正己烷淋洗,丙酮-二氯甲烷（1：1,v/v）洗脱。色谱分离以50 mmol/L NaH₂PO₄-乙腈（7：3,v/v）为流动相,方法的线性范围为0.01~1.00 mg/L,线性相关系数大于0.998,检出限（LOD）为0.003 mg/L,定量限（LOQ）为0.01 mg/L。在0.02~0.2 mg/kg添加范围内,9种磺胺类药物的回收率高于70%,RSD低于8%,表明多壁碳纳米管对磺胺类药物具有较强的吸附富集能力。该方法简便、准确可用于动物组织及产品中磺胺药物残留的检测。