In this study, a Raman immunoprobe with a gold-core silver-shell as the enhanced substrate was prepared, and the simultaneous detection of spiramycin and tilmicosin was achieved through optimizing the key factors, including antibody addition, coagulant type and dosage, concentration of encapsulated antigen, and probe addition.In the method, both spiramycin and tilmicosin exhibited a good linear relationship between the logarithmic values of concentration and inhibition rate within the range of 0.000 1 ng/mL to 10 ng/mL.The detection limits for spiramycin and tilmicosin were determined to be 0.57 pg/mL and 0.25 pg/mL, respectively.The feasibility of the method was validated by conducting spiked experiments in milk, and it was found that the recoveries and relative standard deviations of spiramycin and tilmicosin complied with the requirements of the method.Furthermore, cross-reactivity studies demonstrated specificity, showing no cross-reactivity between spiramycin, tilmicosin, and other antibiotics such as erythromycin, oleandomycin, clarithromycin, or roxithromycin.Overall, the method is rapid, convenient and sensitive, and can meet the requirement of simultaneous determination of spiramycin and tilmicosin.
[1] ALTAN F, YESILMEN ALP S.Antimicrobial susceptibility testing of Arcobacter butzleri and Arcobacter cryaerophilus isolated from buffalo milk with subclinical mastitis:A different approach[J].Journal of the Hellenic Veterinary Medical Society, 2022, 73(2):4227-4234.
[2] XIONG J, ZHU Q, YANG S, et al.Comparison of pharmacokinetics of tilmicosin in healthy pigs and pigs experimentally infected with Actinobacillus pleuropneumoniae[J].New Zealand Veterinary Journal, 2019, 67(5):257-263.
[3] ZHANG Q Q, YING G G, PAN C G, et al.Comprehensive evaluation of antibiotics emission and fate in the river basins of China:Source analysis, multimedia modeling, and linkage to bacterial resistance[J].Environmental Science & Technology, 2015, 49(11):6772-6782.
[4] GAO J, ZHU M M, HUANG H, et al.Advances, challenges and promises of carbon dots[J].Inorganic Chemistry Frontiers, 2017, 4(12):1963-1986.
[5] COLLABORATORS A R.Global burden of bacterial antimicrobial resistance in 2019:A systematic analysis[J].The Lancet, 2022, 399(10325):629-655.
[6] World Health Organization.WHO list of critically important antimicrobials for human medicine (WHO CIA list)[R].World Health Organization, 2019.
[7] LEKE A Z, DOLK H, LOANE M, et al.Macrolide and lincosamide antibiotic exposure in the first trimester of pregnancy and risk of congenital anomaly:A European case-control study[J].Reproductive Toxicology, 2021, 100:101-108.
[8] VEENHUIZEN M F, WRIGHT T J, MCMANUS R F, et al.Analysis of reports of human exposure to Micotil 300 (tilmicosin injection)[J].Journal of the American Veterinary Medical Association, 2006, 229(11):1737-1742.
[9] Official Journal of the European Union.Commission regulation (EU) No.37/2010 on pharmacologically active substances and their classification regarding maximumresidue limits in foodstuffs of animal origin[S].Brussels:European Commission, 2009.
[10] The Japan Food Chemical Research Foundation.Maximum residue limits (MRLs) list of agricultural chemicals in foods[EB/OL].https://db.ffcr.or.jp/front/,2023-03-28.
[11] PEREIRA B F M, PEREIRA M U, FERREIRA R G, et al.Dietary exposure assessment to macrolide antimicrobial residues through infant formulas marketed in Brazil[J].Food Additives & Contaminants:Part A, 2021, 38(10):1672-1688.
[12] SALEH H, ELHENAWEE M, HUSSIEN E M, et al.Validation of HPLC-UV multi-residue method for the simultaneous determination of tetracycline, oxytetracycline, spiramycin and neospiramycin in raw milk[J].Food Analytical Methods, 2021, 14(1):36-43.
[13] KWIECIEN＇ A, KRZEK J, GADEK M.Simultaneous identification and quantitative determination of azithromycin, clarithromycin, roxithromycin, spiramycin and troleandomycin by thin-layer chromatography and densitometry[J].Acta Chromatographica, 2014, 26(4):657-670.
[14] THANH XUAN D, HOANG V D.Application of Fourier transform-based algorithms to resolve spectral overlapping for UV spectrophotometric co-assay of spiramycin and metronidazole in tablets[J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2022, 277:121253.
[15] 洪茜. 基于表面增强拉曼光谱的鸭肉中大环内酯类抗生素残留检测研究[D].南昌:江西农业大学, 2015.
HONG Q. Detection of macrolide antibiotic residues in duck meat based on surface enhanced Raman spectroscopy. Nanchang: Jiangxi Agricultural University, 2015.
[16] LI X M, WEN K, CHEN Y Q, et al.Multiplex immunogold chromatographic assay for simultaneous determination of macrolide antibiotics in raw milk[J].Food Analytical Methods, 2015, 8(9):2368-2375.
[17] JIANG W X, ZHANG H Y, LI X M, et al.Monoclonal antibody production and the development of an indirect competitive enzyme-linked immunosorbent assay for screening spiramycin in milk[J].Journal of Agricultural and Food Chemistry, 2013, 61(46):10925-10931.
[18] SONG Y H, SONG S S, LIU L Q, et al.Simultaneous detection of tylosin and tilmicosin in honey using a novel immunoassay and immunochromatographic strip based on an innovative hapten[J].Food and Agricultural Immunology, 2016, 27(3):314-328.
[19] FRENS G.Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions[J].Nature Physical Science, 1973, 241:20-22.
[20] WU X C, YIN L M, GAO S P, et al.Core-satellite nanoassembly system with aptamer-conjugated Au@ Ag nanoparticles for SERS detection of patulin in apples[J].Food Control, 2024, 159:110293.
[21] HUANG W, GUO E H, LI J G, et al.Quantitative and ultrasensitive detection of brombuterol by a surface-enhanced Raman scattering (SERS)-based lateral flow immunochromatographic assay (FLIA) using AgMBA@Au-Ab as an immunoprobe[J].The Analyst, 2021, 146(1):296-304.
[22] SHI Q Q, HUANG J, SUN Y N, et al.A SERS-based multiple immuno-nanoprobe for ultrasensitive detection of neomycin and quinolone antibiotics via a lateral flow assay[J].Mikrochimica Acta, 2018, 185(2):84.
[23] ZHANG Y X, WU G, WEI J T, et al.Rapid and sensitive detection of rotavirus by surface-enhanced Raman scattering immunochromatography[J].Mikrochimica Acta, 2021, 188(1):3.
[24] ZHOU X X, CHEN S H, PAN Y, et al.High-performance Au@Ag nanorods substrate for SERS detection of malachite green in aquatic products[J].Biosensors, 2023, 13(8):766.
[25] DENG D D, YANG H, LIU C, et al.Ultrasensitive detection of Sudan I in food samples by a quantitative immunochromatographic assay[J].Food Chemistry, 2019, 277:595-603.
[26] BI S Y, YUAN Y, ZHANG F M, et al.A sensitive surface-enhanced Raman spectroscopy detection for gentamicin and tobramycin using γ-Al2O3-modified silver nanoparticles coated with bovine serum albumin as substrate[J].Talanta, 2023, 260:124635.
[27] CHEN Y K, YANG S X, SHI X R, et al.Au@ 4-MBA@ Ag NPs labeled SERS lateral flow immunoassay for ultrasensitive and quantitative detection of Salmonella enteritidis[J].Microchemical Journal, 2023, 193:109134.
[28] LIU Z W, CHEN J Y, ZHAO S J, et al.Immunochromatographic assays based on three kinds of nanoparticles for the rapid and highly sensitive detection of tylosin and tilmicosin in eggs[J].Mikrochimica Acta, 2021, 189(1):42.
[29] YANG J Y, PAN M F, LIU K X, et al.Core-shell AuNRs@Ag-enhanced and magnetic separation-assisted SERS immunosensing platform for amantadine detection in animal-derived foods[J].Sensors and Actuators B:Chemical, 2021, 349:130783.
[30] XU L J, LEI Z C, LI J X, et al.Label-free surface-enhanced Raman spectroscopy detection of DNA with single-base sensitivity[J].Journal of the American Chemical Society, 2015, 137(15):5149-5154.
[31] HU R, ZHANG K N, WANG W, et al.Quantitative and sensitive analysis of polystyrene nanoplastics down to 50 nm by surface-enhanced Raman spectroscopy in water[J].Journal of Hazardous Materials, 2022, 429:128388.
[32] HENDRICKSON O D, ZVEREVA E A, ZHERDEV A V, et al.Development of a double immunochromatographic test system for simultaneous determination of lincomycin and tylosin antibiotics in foodstuffs[J].Food Chemistry, 2020, 318:126510.
[33] WANG Z H, LI H, LI C L, et al.Development and application of a quantitative fluorescence-based immunochromatographic assay for fumonisin B1 in maize[J].Journal of Agricultural and Food Chemistry, 2014, 62(27):6294-6298.
[34] LI X M, SHEN J Z, WANG Q, et al.Multi-residue fluorescent microspheres immunochromatographic assay for simultaneous determination of macrolides in raw milk[J].Analytical and Bioanalytical Chemistry, 2015, 407(30):9125-9133.
