Determination of metoserpate,buquinolate, and diclofenac in pork,eggs, and milk using liquid chromatography–tandem mass spectrometry

In this work, a method was developed for the simultaneous determination of residual metoserpate, buquinolate and diclofenac in pork, milk, and eggs. Samples were extracted with 0.1% formic acid in acetonitrile, defatted with n‐hexane, and filtered prior to analysis using liquid chromatography–tandem mass spectrometry. The analytes were separated on a C₁₈ column using 0.1% acetic acid and methanol as the mobile phase. The matrix‐matched calibration curves showed good linearity over a concentration range of 5–50 ng/g with coefficients of determination (R²) ≥0.991. The intra‐ and inter‐day accuracies (expressed as recovery percentage values) calculated using three spiking levels (5, 10, and 20 μg/kg) were 80–108.65 and 74.06–107.15%, respectively, and the precisions (expressed as relative standard deviation) were 2.86–13.67 and 0.05–11.74%, respectively, for the tested drugs determined in various matrices. The limits of quantification (1 and 2 μg/kg) were below the uniform residual level (0.01 mg/kg) set for compounds that have no specific maximum residue limit (MRL). The developed method was tested using market samples and none of the target analytes was detected in any of the samples. The validated method proved to be practicable for detection of the tested analytes in pork, milk, and eggs.     

Residue analysis of tebufenozide and indoxacarb in chicken muscle,milk, egg and aquatic animal products using liquid chromatography–tandem mass spectrometry

We developed an analytical method using liquid–liquid extraction (LLE) and liquid chromatography–tandem mass spectrometry (LC‐MS/MS) to detect and quantify tebufenozide (TEB) and indoxacarb (IND) residues in animal and aquatic products (chicken muscle, milk, egg, eel, flatfish, and shrimp). The target compounds were extracted using 1% acetic acid (0.1% acetic acid for egg only) in acetonitrile and purified using n‐hexane. The analytes were separated on a Gemini‐NX C₁₈ column using (a) distilled water with 0.1% formic acid and 5 mm ammonium acetate and (b) methanol with 0.1% formic acid as the mobile phase. All six‐point matrix‐matched calibration curves showed good linearity with coefficients of determination (R²) ≥0.9864 over a concentration range of 5–50 μg/kg. Intra‐ and inter‐day accuracy was expressed as the recovery rate at three spiking levels and ranged between 73.22 and 114.93% in all matrices, with a relative standard deviation (RSD, corresponding to precision) ≤13.87%. The limits of quantification (LOQ) of all target analytes ranged from 2 to 20 μg/kg, which were substantially lower than the maximum residue limits (MRLs) specified by the regulatory agencies of different countries. All samples were collected from different markets in Seoul, Republic of Korea, and tested negative for tebufenozide and indoxacarb residues. These results show that the method developed is robust and may be a promising tool to detect trace levels of the target analytes in animal products.     

Simultaneous determination of multiveterinary drug residues in pork meat by liquid chromatography‐tandem mass spectrometry combined with solid phase extraction

An LC‐MS/MS method developed for simultaneous analysis of 54 veterinary drug residues of six families in pork meat samples, including sulfanilamide, nitroimidazoles, quinolones, macrolide antibiotics, lincosamides, and praziquantel. The pork meat sample was prepared by extraction with ACN, and clean‐up on a C₁₈ SPE cartridge. The sample was separated on a C₈ column and eluted with ACN, methanol, and formic acid. The MS/MS detector is operated in the multiple reaction monitoring mode, acquiring two specific precursor‐product ion transitions per target compound. The method showed excellent linearity (R² ≥ 0.99) and high precision (relative SD, RSD ≤ 19.8%) for all compounds. The method quantification limits of 54 veterinary drug residues were in the range of 0.3–3.0 μg/kg. Recoveries for most analytes based on matrix‐matched calibration in matrices were 20.9–121.0%. This method has been successfully applied for analysis of more than 100 pork meat samples from the local market; five of the 54 drugs were detected.     

Simultaneous detection of bacitracin and polymyxin B in livestock products using liquid chromatography with tandem mass spectrometry

With the overarching aim to develop a simple and reliable method for the quantitative analysis of polypeptide antibiotics in various livestock products, the content of bacitracin, and polymyxin B in pork, beef, chicken, milk, and eggs was analyzed using colistin sulfate as an internal standard. The extracted samples were eluted via solid‐phase extraction using 2% formic acid in acetonitrile/methanol (1:1, v/v). The two polypeptides were identified and quantified based on the intensities of mass fragments from the respective triply charged precursor ions (bacitracin: 474.97 amu and polymyxin B: 402 amu) at the defined retention time windows using liquid chromatography with electrospray ionization tandem mass spectrometry in time‐scheduled multiple reaction monitoring mode. The calibration curves showed good linearity over the concentration range 50–2500 ng/mL with determination coefficients ≥ 0.991. The mean recoveries were in the range 80.3–88.8% with relative standard deviations <13% for all samples. The limits of quantitation ranged from 30–250 ng/g. The developed method was applied to market samples, but the target analytes were not detected in any of the samples. The developed method is reliable for the simultaneous detection of bacitracin and polymyxin B in pork, beef, chicken, milk, and eggs.     

Single‐step multiresidue determination of ten multiclass veterinary drugs in pork,milk, and eggs using liquid chromatography with tandem mass spectrometry

A multiclass, multiresidue determination method is reported for the detection of ten veterinary drugs, including scopolamine, metoclopramide, acriflavine, berberine, tripelennamine, diphenhydramine, acrinol, triamcinolone, loperamide, and roxithromycin in pork, milk, and eggs. The method involves a simple extraction using 0.1% formic acid in acetonitrile, followed by defatting with n‐hexane, centrifugation, and filtration prior to liquid chromatography with tandem mass spectrometric analysis. As ion suppression and enhancement effects are reported, matrix‐matched calibrations are used for quantification, with determination coefficients ≥0.9765. For the majority of the tested analytes, the intra‐ and interday accuracy (expressed as recovery %) range from 70.6 to 94.6% and from 70.1 to 93.3%, respectively, and the precision (expressed as relative standard deviation) ranges from 0.5 to 19.8% and from 2.8 to 18.4% in all matrices. The limits of quantification range between 0.5 and 10 ng/g. The validated tandem mass spectrometry method is successfully applied to market samples; the target analytes are not detected in any of the tested samples. In terms of accuracy, no extract cleanup is deemed necessary. The developed method is feasible for the simultaneous detection of the tested analytes in pork, milk, and eggs.     

Simultaneous determination of difenoconazole,trifloxystrobin and its metabolite trifloxystrobin acid residues in watermelon under field conditions by GC–MS/MS

An optimized quick, easy, cheap, effective, rugged and safe method for the simultaneous determination of difenoconazole, trifloxystrobin and its metabolite trifloxystrobin acid residues in watermelon and soil was developed and validated by gas chromatography with tandem mass spectrometry. The samples were extracted with acetonitrile (1% formic acid) and cleaned up by dispersive solid‐phase extraction with octadecylsilane sorbent. The limit of quantification of the method was 0.01 mg/kg, and the limit of detection was 0.003 mg/kg for all three analytes. The recoveries of the fungicides in watermelon, pulp and soil were 72.32–99.20% for difenoconazole, 74.68–87.72% for trifloxystrobin and 78.59–92.66% for trifloxystrobin acid with relative standard deviations of 1.34–14.04%. The dissipation dynamics of difenoconazole and trifloxystrobin in watermelon and soil followed the first‐order kinetics with half‐lives of 3.2–8.8 days in both locations. The final residue levels of difenoconazole and trifloxystrobin were below 0.1 mg/kg (maximum residue level [MRL] set by China) and 0.2 mg/kg (MRL set by European Union), respectively, in pulp samples collected 14 days after the last application. These results could help Chinese authorities to establish MRL of trifloxystrobin in watermelon and provide guidance for the safe and proper application of both fungicides on watermelon.     

Determination of bithionol, bromophen, nitroxynil, oxyclozanide, and tribromsalan in milk with liquid chromatography coupled with tandem mass spectrometry

LC/MS/MS was developed to determine the residues of bithionol (BTN), bromofen (BMF), nitroxynil (NTX), oxyclozanide (OCZ), and tribromsalan (TBS) in milk. Samples were extracted with ethyl acetate and cleaned up by liquid-liquid separation with acetonitrile and n-hexane. The compounds were determined by RP-LC using a C18 column with 0.1% formic acid-methanol. Mass spectral acquisition was performed in the negative mode by applying selected-reaction monitoring. The method was validated in milk spiked with these compounds at 5-600 microg/kg; average recoveries were in the range 83.8-97.1%, with RSD values of 1.4-8.0%. The interassay RSDs were less than 11%. The LODs of these compounds in milk were 0.1 microg/kg. The method was applied to 24 raw milk samples. The concentration of these compounds in all samples was lower than the Japanese maximum residue limits. The method is rapid, sensitive, and specific for monitoring residues of BTN, BMF, NTX, OCZ, and TBS in milk.     

Residues,dissipation and risk evaluation of spiroxamine in open-field-grown strawberries using liquid chromatography tandem mass spectrometry

The dissipation dynamic and residues of spiroxamine in open-field-grown strawberries were determined using liquid chromatography tandem mass spectrometry (LC–MS/MS). Spiroxamine application was performed according to Egyptian good agricultural practices recommendation. A QuEChERS-based extraction method along with direct analysis with an LC–MS/MS analytical method were optimized and validated, and the specificity of the techniques used was considered satisfactory. Good linearity (R² > 0.999) was obtained for spiroxamine within the range of 0.001–0.1 μg/ml. The mean recoveries varied between 97.1 and 108.2%, with inter- and intra-day precision (RSD) <4.9%. The limit of quantitation for spiroxamine was 0.001 mg/kg. The results indicated that spiroxamine degradation in strawberry followed first–order kinetics (R² > 0.9929) with an estimated half-life value of 4.71 days. Considering the Australian maximum residue limit (0.05 mg/kg) in strawberry and based on the results from residue trials with a preharvest interval of 14 days for strawberry, compliance can be expected. The present results could provide guidance to fully evaluate the risks of spiroxamine residues, preventing any potential health risk to consumers.     

Multiresidue LC–MS/MS analysis of cephalosporins and quinolones in milk following ultrasound‐assisted matrix solid‐phase dispersive extraction combined with the quick,easy, cheap,effective, rugged,and safe methodology

A sensitive and selective confirmatory method for milk‐residue analysis of ten quinolones and eight cephalosporins by LC‐MS/MS has been developed herein. For the chromatographic separation of target analytes, a Perfectsil ODS‐2 (250 × 4 mm, 5 μm) analytical column was used and gradient elution was applied, using a mobile phase of 0.1% w/w TFA in water and 0.1% w/w TFA in ACN. Ultrasound‐assisted matrix solid‐phase dispersion procedure was applied for the extraction and clean‐up procedure of antimicrobials agents from milk matrix using a mixture of Bond Elut Plexa sorbent and QuEChERS. The method was validated meeting the European Legislation determining selectivity, linearity response, trueness, precision (repeatability and between‐day reproducibility), decision limit, detection capability, and ruggedness following the Youden approach. Recoveries of all antibiotics ranged from 81.7 to 117.9%, while RSD values were lower than 13.7%. Limits of quantification for all examined compounds ranged from 2.4 to 15.0 μg/kg, substantially lower than the maximum residue limits established by the European Union (30–100 μg/kg).     

Monitoring pesticide residues in dates marketed in Al‐Qassim,Saudi Arabia using a QuEChERS methodology and liquid chromatography–tandem mass spectrometry

A sensitive, simple and rapid QuEChERS extraction method and liquid chromatography equipped with triple quadrupole mass spectrometry (LC‐MS/MS) were used to determine 42 pesticides in dates. Acidified acetonitrile and citrate buffer salts were used to extract re‐hydrated samples. Acceptable validation performances were achieved, i.e. recovery range of 70−120% and RSD values ≤20% for 42 analytes at three different concentrations:100, 50 and 10 μg/kg. This method was used to analyse 200 date fruit samples (var. Sukkari) collected from different large markets in the Al‐Qassim region in Saudi Arabia. Pesticide residues were detected in 36 (18%) of the date fruits samples, and 15 samples (7.5%) exceeded the maximum residue levels. The ruggedness test results showed that this method is robust and suitable for the determination of pesticide residues in dates. Additionally, the results showed that the monitored dates did not have a health impact on consumers in Saudi Arabia during the study period.     

Effects of mineral oil spray additives on the distribution and dissipation kinetics of pyraclostrobin and azoxystrobin in banana leaves,fruits, and soil

Using LC–MS/MS, a rapid and sensitive method for the simultaneous determination of pyraclostrobin and azoxystrobin residues in banana matrices (leaf and whole banana) and soil was established. The samples were extracted using acetonitrile and purified through C₁₈ dispersive solid-phase extraction. The average recovery of the analytes in various matrices was in the range of 77.3%–103.9% with an RSD range of 0.9%–9.5%. The initial deposition amounts of pyraclostrobin and azoxystrobin at 2 h in the banana leaves of the mineral oil group were 1.43 and 1.31 times in Guangxi, and 2.10 and 1.81 times in Hainan for the water group, whereas those in the soil of the water group were 3.45 and 3.03 times in Guangxi, and 2.14 and 3.48 times in Hainan for the mineral oil group. The half-lives in the leaves and soil of the mineral oil group were not remarkably different from those of the water group. The terminal residue of the analytes on the whole banana was <0.02 mg/kg at 14 days after application from the two sites. The results of this work may indicate and promote the safety of using pyraclostrobin and azoxystrobin in banana production, especially with mineral oil spray adjuvants.     

Sensitive monitoring of penicillin antibiotics in milk and honey treated by stir bar sorptive extraction based on monolith and LC‐electrospray MS detection

In the present study, a convenient and sensitive method for determination of six penicillin antibiotics (amoxicillin, ampicillin, penicillin G, oxacillin, cloxacillin, and dicloxacillin) in milk and honey samples was developed. Milk and honey samples were diluted with water, then directly treated by stir bar sorptive extraction based on poly (vinylimidazole‐divinylbenzene) monolithic material as coating. The analytes were analyzed by LC/ESI‐ MS/MS. Several extraction parameters including extraction and desorption time, pH value, and ionic strength in sample matrix were investigated in detail. Under the optimized extraction conditions, the calculated detection limits for the target compounds were as low as 0.23–2.66 ng/kg in milk and 0.18–1.42 ng/kg in honey, respectively. Good linearity was obtained for analytes with the correlation coefficients (R²) above 0.997. Excellent method reproducibility was achieved in terms of intraday and interday precisions, indicated by the RSDs of <5.0 and <10.0%, respectively. Finally, the proposed method was successfully applied to the determination of penicillin antibiotics residues in different milk and honey samples.     

液相色谱串联质谱法测定猪肉和猪肝中9种磺胺类药物的残留

建立了高效液相色谱-串联质谱法(HPLC/MS/MS)同时测定猪肉及猪肝中9种磺胺类药物残留的检测方法.样品经10％的Na2SO4溶液和乙腈-氯仿(10:1)提取,乙腈饱和正己烷去脂,使用乙二胺-N-丙基硅烷(PSA)和十八烷基键合相硅胶(ODS C18-N)两种基质分散净化剂净化,采用LC-MS/MS多反应监测(MRM)正离子模式测定,内标法定量.9种磺胺检出限为0.1 ～0.8 μg/kg,在5,10,20μg/kg 3个浓度添加水平,回收率为74.1％ ～ 115.8％,相对标准偏差均小于6.2％(n=6).     

Determination,residue and risk assessment of trifloxystrobin,trifloxystrobin acid and tebuconazole in Chinese rice consumption

A simple and rapid analytical method for the detection of trifloxystrobin, trifloxystrobin acid and tebuconazole in soil, brown rice, paddy plants and rice hulls was established and validated by liquid chromatography with tandem mass spectrometry. Acceptable linearity (R² > 0.99), accuracy (average recoveries of 74.3–108.5%) and precision (intra- and inter-day relative standard deviations of 0.9–8.8%) were obtained using the developed determination approach. In the field trial, the half-lives of trifloxystrobin and tebuconazole in paddy plants were 5.7–8.3 days in three locations throughout China, and the terminal residue concentrations of trifloxystrobin and tebuconazole were <100 and 500 μg/kg (maximum residue limits set by China), respectively, at harvest, which indicated that, based on the recommended application procedure, trifloxystrobin and tebuconazole are safe for use on rice. The risk assessment results demonstrated that, owing to risk quotient values of both fungicides being <100%, the potential risk of trifloxystrobin and tebuconazole on rice was acceptable for Chinese consumers. These data could provide supporting information for the proper use and safety evaluation of trifloxystrobin and tebuconazole in rice.     

Determination of tetracycline antibiotics in cow's milk by liquid chromatography/tandem mass spectrometry

A liquid chromatographic/tandem mass spectrometric (LC/MS/MS) multiresidue method for the simultaneous quantitative determination of oxytetracycline, 4-epi-oxytetracycline, tetracycline, 4-epi-tetracycline, chlortetracycline, 4-epi-chlortetracycline and doxycycline in milk has been developed. An extraction procedure consisting of a liquid extraction of the milk samples with trichloroacetic acid was performed. The extract was centrifuged and the supernatant was filtered. Solid-phase extraction (SPE) with an OASIS HLB SPE column was used to clean up the sample extracts. The samples were analysed by LC/MS/MS. The LC separation was performed on a reversed-phase C18 column using gradient elution with a mobile phase consisting of water and a mixture of methanol/acetonitrile. The tetracycline analytes were detected with a quadrupole mass spectrometer using positive ion electrospray ionisation. The confirmatory method has acceptable detection limits and the different tetracyclines can be detected at a residue concentration between 5 and 20 microg/L. The method is validated according to the European requirements for veterinary drug residues and all determined parameters were found to conform to the criteria. The recovery values ranged from 90.4 to 101.2% with relative standard deviations (RSDs) no larger than 9.7%. The overall or between-day precision of the analytical assay determined as repeatability at several residue concentrations and expressed as RSD ranged from 3.3 to 10%. This analytical assay is a useful tool within the Belgian monitoring programme for confirmation of samples which have been positively screened for residues of tetracyclines in raw farm cow's milk.     

Quantitative analysis of penicillins in porcine tissues, milk and animal feed using derivatisation with piperidine and stable isotope dilution liquid chromatography tandem mass spectrometry

Penicillins are used universally in both human and veterinary medicine. The European Union (EU) has established maximum residue levels (MRLs) for most ß-lactam antibiotics in milk and animal tissues and included them in the National Residue Monitoring Programs. In this study, a novel method is described for the determination and confirmation of eight penicillins in porcine tissues, milk and animal feed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). To prevent degradation of penicillin residues during workup, a derivatisation procedure was developed, by which penicillins were converted to stable piperidine derivatives. Deuterated piperidine derivatives were synthesised for all relevant penicillins, enabling the use of isotope dilution for accurate quantification. Penicillin residues were derivatised in the crude extract with piperidine and isolated using solid-phase extraction. The penicillin piperidine derivatives were determined by LC–MS/MS. The method was validated at the current MRLs, which range from 25–300 µg kg^?1 in muscle and kidney to 4–30 µg kg^?1 in milk as well as at the target value of 100 µg kg^?1 chosen for animal feed, according to the EU requirements for a quantitative confirmatory method. Accuracy ranged from 94–113% (muscle), 83–111% (kidney) and 87–103% (milk) to 88–116% (animal feed). Intra-day precision (relative standard deviation (RSD)_r) ranged from 5–13% (muscle, n?=?18), 4–17% (kidney, n?=?7) and 5–18% (milk, n?=?7) to 11–32% (animal feed, n?=?18). Inter-day precision (RSD_RL, n?=?18) ranged from 6–23% (muscle) to 11–36% (animal feed). From the results, it was concluded that the method was fit for purpose at the target MRLs in animal tissue and target levels for animal feed.     

Simultaneous determination of residues of chloramphenicol,thiamphenicol, florfenicol,and florfenicol amine in farmed aquatic species by liquid chromatography/mass spectrometry

A liquid chromatographic (LC)/mass spectrometric (MS) method was developed for determining the residues of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in a number of aquatic species. The phenicols are extracted with acetone, the extracts are partitioned with dichloromethane, the aqueous layer is removed, and the organic layer is evaporated to dryness. The residue is dissolved in dilute acid and defatted with hexane, and the aqueous layer is prepared for analysis by LC. The phenicols are determined by reversed-phase LC by using a Hypersil C18-BD column with a water-acetonitrile gradient and MS detection using selected-ion recording. Calibration curves were linear for all analytes between 0.015 and 0.425 ng injected. The relative standard deviations for measurements by the proposed method were < 10% for all of the analytes studied, with recoveries ranging from 71% for florfenicol amine to 107% for florfenicol in salmon tissue spiked at the 2 ng/g level. Detection limits of 0.1 ng/g for florfenicol and chloramphenicol, 0.3 ng/g for thiamphenicol, and 1.0 ng/g for florfenicol amine are easily obtainable. The operational errors, interferences, and recoveries for spiked samples compare favorably with those obtained by established LC methodology. The proposed method is simple, rapid, and specific for monitoring residues of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in a number of aquatic species.     

高效液相色谱-串联质谱法测定猪肉中的阿维菌素类、地克珠利、妥曲珠利及其代谢物残留

建立了猪肉中阿维菌素、伊维菌素、多拉菌素、莫西菌素、乙酰氨基阿维菌素等5种阿维菌素类药物以及地克珠利、妥曲珠利、妥曲珠利砜、妥曲珠利亚砜等4种均三嗪类药物及其代谢物残留的高效液相色谱-串联质谱分析方法。样品采用乙腈提取,ODS粉分散固相萃取净化,经Venusil ASB C18色谱柱(150 mm×2.1 mm, 3.0 μm)分离,电喷雾串联四极杆质谱多反应离子监测方式测定。9种分析物在0.005～0.2 mg/L范围内线性关系良好,相关系数均大于0.990。猪肉基质中9种分析物在0.005、0.01、0.02 mg/kg加标水平下,回收率为73.2%～91.5%,相对标准偏差为12%～17%。该方法稳定、可靠,可满足猪肉中阿维菌素类、均三嗪类药物残留的检测与确证。     

超高效液相色谱-串联质谱法测定猪肉、鸡蛋、牛奶中9种食源性兴奋剂类药物残留

β-受体激动剂、β-阻断剂、蛋白同化制剂属于兴奋剂类药物,在动物饲养和屠宰过程中的违禁使用成为食源性兴奋剂类药物残留的来源,危害人类健康.目前 β-受体激动剂、蛋白同化制剂的检测较多,β-阻断剂检测报道较少,动物源食品中 β-阻断剂检测尚无标准方法.该文建立了超高效液相色谱-串联质谱测定猪肉、鸡蛋、牛奶中 β-受体激动...     

Effect of background derivatization on the signal enhancement of pesticide residues extracted from edible oils

The effect of background derivatization on the signal enhancement of pesticide residues extracted from edible oil samples was studied by GC with negative chemical ionization MS. The analytes were extracted by a solvent extraction process, and the extract was subjected to rapid low‐temperature fat precipitation. The residual fatty acids were silylated by derivatization with N,O‐bis(trimethylsilyl)trifluoroacetamide. The chromatograms obtained from the derivatized samples showed higher signal intensity and lower detection levels when compared to the direct analysis without derivatization. The sensitivity levels of the method are either better or comparable to that of previously reported methodologies. The LODs of the analyzed organochlorine, organophosphorus, and synthetic pyrethroid residues in sunflower, rice bran, and ground oil samples were in the range of 0.02–0.5 ng/g, and the LOQs were in the range of 0.1–2 ng/g. The intraday and interday accuracies were in the range of 81–116% with RSDs less than 14%. The recoveries obtained were in the range of 53–89% with the RSD values less than 13% for all the studied pesticide residues.