该文利用液相色谱-串联质谱建立了同时检测虾肉中安眠酮、地西泮、诺氟沙星以及氧氟沙星的分析方法,并且用此分析方法探究这4种兽药的热稳定性。虾肉样品用乙腈和无水硫酸钠提取,经过C18固相萃取柱净化,以Eclipse XDB-C18柱分离,在正离子模式下用多反应监测模式进行数据采集,基质外标法进行定量。此外,在90 ℃水浴加热条件下,对4种兽药进行热稳定性研究,加热15、30、45、60 min后,以该文建立的分析方法进行检测。4种兽药在0.05~40 ng/mL线性范围内,相关系数R2均大于0.99,检出限为0.01~0.38 μg/kg,4种化合物的回收率为78%~119%,相对标准偏差为1.14%~4.76% 。该方法灵敏度高、准确性好,适用于虾肉中安眠酮、地西泮、诺氟沙星以及氧氟沙星的同时检测,并且研究表明,这4种兽药具有良好的热稳定性,在水煮加热过程中不会分解或者转化。
An analytical method was established to simultaneously determine methaqualone, diazepam, norfloxacin, and ofloxacin in shrimp by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and the thermal stability of the four veteri- nary drugs were studied. The samples were extracted with a mixture of anhydrous sodium sulfate and acetonitrile and purified by C18 car- tridge solid-phase extraction (SPE). Eclipse XDB-C18 column (2.1 mm×150 mm, 3.5 μm) was utilized to separate analytes. All veterinary drug detection was performed on multiple reaction monitoring (MRM) mode in the positive ion mode and the matrix external standard method was for quantification. Furthermore, under the condition of 90 ℃ water bath heating, the thermal stability of the four veterinary drugs was studied. After heating for 15, 30, 45, and 60 min, the analytical method established in this research was used for detection. Results demonstrated that all four veterinary drugs had satisfactory linearity in the concentration range of 0.05-40 ng/mL with correlation coefficients greater than 0.99. The limits of detection (LOD) were between 0.01 μg/kg and 0.38 μg/kg. The average recovery rate was 78%-119% and the relative standard deviation was1.14%-4.76%. This method is sensitive and accurate, which is suitable for the simultaneous detection of methaqualone, diazepam, norfloxacin, and ofloxacin in shrimp. Moreover, the four veterinary drugs have satisfactory thermal stability, and will not decompose or transform during heating.
[1] FISHERIES AND AQUACULTURE DEPARTMENT.The State of World Fisheries and Aquaculture 2012[M].Rome:FAO,2012.
[2] ZHANG R L, KANG Y R, ZHANG R J, et al.Occurrence, source, and the fate of antibiotics in mariculture ponds near the Maowei sea, South China:Storm caused the increase of antibiotics usage[J].Sci-ence of the Total Environment, 2021, 752:141882.
[3] HAN D, SHAN X J, ZHANG W B, et al.A revisit to fishmeal usage and associated consequences in Chinese aquaculture[J].Reviews in Aquaculture, 2018, 10(2):493-507.
[4] HE K, BLANEY L.Systematic optimization of an SPE with HPLC-FLD method for fluoroquinolone detection in wastewater[J].Journal of Hazardous Materials, 2015, 282:96-105.
[5] RUSU A, HANCU G, UIVAROŞI V.Fluoroquinolone pollution of food, water and soil, and bacterial resistance[J].Environmental Chemistry Letters, 2015, 13(1):21-36.
[6] YU K L, YUE M E, XU J, et al.Determination of fluoroquinolones in milk, honey and water samples by salting out-assisted dispersive liquid-liquid microextraction based on deep eutectic solvent combined with MECC[J].Food Chemistry, 2020, 332:127371.
[7] ZAYED M A, FAHMEY M A, HAWASH M F.Investigation of diazepam drug using thermal analyses, mass spectrometry and semi-empirical MO calculation[J].Spectrochimica Acta Part A:Molecular & Biomolecular Spectroscopy, 2005, 61(5):799-805.
[8] CABRERA G C, WAISBAUM R G D, NUDELMAN N S.Kinetic and mechanistic studies on the hydrolysis and photodegradation of diazepam and alprazolam[J].Journal of Physical Organic Chemistry, 2005, 18(2):156-161.
[9] ZHANG L J, ZHANG S F, WAN Y Z.Voltammetric behavior of methaqualone and its determination by single-sweep oscillopolarography[J].Talanta, 2003, 59(5):1 009-1 013.
[10] ZHANG L Q, WU P G, ZHANG Y M, et al.A GC/MS method for the simultaneous determination and quantification of chlorpromazine and diazepam in pork samples[J].Analytical Methods, 2014, 6(2):503-508.
[11] VÁZQUEZ M M P, VÁZQUEZ P P, GALERA M M, et al.Determination of eight fluoroquinolones in groundwater samples with ultrasound-assisted ionic liquid dispersive liquid-liquid microextraction prior to high-performance liquid chromatography and fluorescence detection[J].Analytica Chimica Acta, 2012, 748:20-27.
[12] WU H, LIU Y L, CHANG J Z, et al.Extraction of five fluoroquinolones in eggs by magnetic solid-phase extraction with Fe3O4-MoS2 and determination by HPLC-UV[J].Food Analytical Methods, 2019, 12(3):712-721.
[13] FERNÁNDEZ P, LAFUENTE N, BERMEJO A M, et al.HPLC determination of cocaine and benzoylecgonine in plasma and urine from drug abusers[J].Journal of Analytical Toxicology, 1996, 20(4):224-228.
[14] BAGHERI N, AI LAWATI H A J, AI SHARJI N A, et al. Mag-etic zinc based 2D-metal organic framework as an efficient adsorbent for simultaneous determination of fluoroquinolones using 3D printed microchip and liquid chromatography tandem mass spectrometry[J].Talanta, 2021, 224:121796.
[15] BANASZKIEWICZ L, WOZ' NIAK M K, KATA M, et al.Rapid and simple multi-analyte LC-MS/MS method for the determination of benzodiazepines and Z-hypnotic drugs in blood samples:Development, validation and application based on three years of toxico- logical analyses[J].Journal of Pharmaceutical and Biomedical Analysis, 2020, 191:113569.
[16] WANG H, ZHAO X Y, XU J W, et al.Determination of quinolones in environmental water and fish by magnetic metal organic frameworks based magnetic solid-phase extraction followed by highperformance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A, 2021, 1651:462286.
[17] SOFALVI S, LAVINS E S, KASPAR C K, et al.Development and validation of an LC-MS-MS method for the detection of 40 benzodiazepines and three Z-drugs in blood and urine by solid-phase extrac- tion[J].Journal of Analytical Toxicology, 2020, 44(7):708-717.
[18] DE BAIRROS A D, DE ALMEIDA R D, PANTALEÃO L, et al.Determination of low levels of benzodiazepines and their metabolites in urine by hollow-fiber liquid-phase microextraction(LPME)and gas chromatography-mass spectrometry(GC-MS)[J].Journal of Chromatography B:Analytical Technologies in the Biomedical and Life Sciences, 2015, 975:24-33.
[19] CHENG L L, ZHANG Y J, SHEN J Z, et al.GC-MS method for simultaneous determination of four sedative hypnotic residues in swine tissues[J].Chromatographia, 2010, 71(1-2):155-158.
[20] FAMIGLINI G, CAPRIOTTI F, PALMA P, et al.The rapid measurement of benzodiazepines in a milk-based alcoholic beverage using QuEChERS extraction and GC-MS analysis[J].Journal of Analytical Toxicology, 2015, 39(4):306-312.
[21] 刘家阳, 黄旭, 贾宏新.固相萃取/超高效液相色谱-串联质谱法测定畜肉中16种镇静剂类兽药残留[J].分析测试学报, 2017, 36(3):305-311.
LIU J Y, HUANG X, JIA H X.Simultaneous determination of 16 sedative residues in livestock products by solid phase extraction/ultra performance liquid chromatography-tandem mass spectrometry[J].Journal of Instrumental Analysis, 2017, 36(3):305-311.
[22] 王綪.贮藏和加工工艺对牛肉品质及氟喹诺酮类药物残留的影响[D].天津:天津农学院, 2015.
WANG Q.Effects of storage and processing technic on quality and fluoroquinolones residues in beef[D].Tianjin:Tianjin Agricultural University, 2015.
[23] 霍晓荣.加工过程对鸡肉和鸡蛋中盐霉素和莫能菌素残留影响的研究[D].南京:南京农业大学, 2006.
HUO X R.Study on effect of processing on salinomycin and monensin residues in eggs and chicken[D].Nanjing:Nanjing Agricultural University, 2006.
