Microsoft Word 王婵-校对 72 不要动版

Size: px

Start display at page:

Download "Microsoft Word 王婵-校对 72 不要动版"

慕哀全
5 years ago
Views:

1 6 1 Vol. 6 No Journal of Food Safety and Quality Jan., 2015 QuEChERS 王婵 1,2, 赵永拓 3, 彭心婷 4, 孙兴权 2, 刘慧颖 2, 张彧 1, 曹际娟 2* (1., ; 2., ; 3., ; 4., ) 摘要 : 目的 QuEChERS (domoic acid, DA) 方法 QuEChERS, : : =6:3:1(v:v:v), C18,, 242 nm, 结果 (limit of quantitation, LOQ) 1.8 μg/g, DA 0.72~30.00 μg/ml ( r=0.9998), 1.8~6.0 μg/g, 90%, (relative standard deviation, RSD) 5% 结论 DA 20 μg/g,, 关键词 : ; QuEChERS ; ; Rapid detection of domoic acid in scallops by QuEChERS method coupled with high performance liquid chromatography WANG Chan 1,2, ZHAO Yong-Tuo 3, PENG XIN-Ting 4, SUN Xing-Quan 2, LIU Hui-Ying 2, ZHANG Yu 1, CAO Ji-Juan 2* (1. Dalian Polytechnic University, Dalian , China; 2. Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian , China; 3. Bayuquan Entry-Exit Inspection and Quarantine Bureau, Yingkou , China; 4. Tacheng Entry-Exit Inspection and QuarantineBureau, Tacheng , China) ABSTRACT: Objective To established a method to detect domoic acid residues in scallops by the sample processing method of QuEChERS in combination with high performance liquid chromatography (HPLC). Methods Samples were cleaned up by QuEChERS with extracting by methanol : acetonitrile : water = 6:3:1 (v:v:v), and purified by C18 powder, separated by reversed phase high performance liquid chromatography and detected at wave length of 242 nm, and quantified by the matrix matching calibration curve external standard method of matrix matching calibration curve at last. Results The limit of quantitative was 1.8 μg/g, working curve correlation coefficient was at the linear range of 0.72~30.00 μg/ml, the recovery rate was higher than 90% at the spiking concentration range of 1.8~6.0 μg/g, and relative standard deviation (RSD) was less than 5%. Conclusion This method with a fast speed and a high sensitivity and exactitude can satisfy the limit 基金项目 : ( ) Fund: Supported by the Special Scientific Research Fund of AQSIQ Public Welfare Profession of China ( ) * 通讯作者 :,,, cjj0909@163.com *Corresponding author: CAO Ji-Juan, Researcher, Liaoning Entry-Exit Inspection and Quarantine Bureau, No.60, Changjiang East Road, Dalian , China. cjj0909@163.com

2 1, : QuEChERS 73 of detection of DA 20 μg/g. KEY WORDS: domoic acid; QuEChERS extration; high performance liquid chromatography; rapid detection 1 引言,, (paralytic shellfish poison, PSP)(diarrhetic shellfish poisions, DSP)(neurotoxic shellfish poisoning, NSP) (amnesic shellfish poisoning, ASP)4 ASP (domoic acid, DA), 1987 [1], DA, [2-4],, DA,,,, 1 DA,, Na +, Na +, Ca 2+,,,, [5] DA 20 μg/g, [6] 1 Fig. 1 Structure of domoic acid DA [7] [8,9] [10,11] [12,13], [14-16],, QuEChERS(quick, eqsy, cheap, effective, rugged and safe),,, DA QuEChERS,,, (primarysecondary amine, PSA) QuEChERS, DA, DA 2 材料和方法 2.1 仪器及试剂 : Agilent 1260( Agilent ); ( Scientific insustries ); ( Sartorius ); ( Thermo ) ( 90%, Sigma ); (, Meker ); ( 98%, ); C18 ( ); (Milli-Q ); 0.22 μm ( Thermo ); : : : =6:3:1 (v:v:v) DA : 10%, DA, 1500 μg/ml; : 5, DA, 2.3.2, DA μg/ml, 0.22 µm 2.2 色谱条件 : Purospher STAR RP-18 (150 mm 4.6

3 74 6 mm 3 μm, Merck ); : 35 ; : 1 ml/min; : 242 nm; : 10 μl; : 0.1% : (87:13, v:v) 2.3 实验方法试样制备,,,,, 100~150 g, 提取与净化 4 g 50 ml, 10 ml : : =6:3:1(v:v:v) 2 g 1 g, 1 min, 5 min, r/min 4 10 min, 4 ml, 0.2 g C18, 0.4 g, 1 min, 8000 r/min 4 10 min, 1 ml 0.22 μm, X=cV/m, X DA (μg/ g); c DA (μg/ml); V (ml); m (g), : : =8:1:1 7:2:1 6:3:1 5:4:1(v:v:v) : =1:1(v:v) : =9:1(v:v) : =17:83(v:v), 1, 1.8 μg/g,, : : =6:3:1(v:v), : : =6:3:1(v:v:v) 0.72 μg/ml DA 2, (1.8 μg/g) 3 2 (0.72 μg/ml) Fig. 2 Chromatography of domoic acid(0.72 μg/ml) 3 结果与讨论 3.1 提取条件的选择提取溶剂的优化 DA, DA, DA,, : =1:1(v:v),,,, DA, QuEChERS, QuEChERS, 3 (1.8 μg/g)da Fig. 3 Chromatography of domoic acid at the low spiking concentration of 1.8 μg/g in scallops 表 1 不同提取溶剂对 DA 回收率的影响 Table 1 Effects of different extraction solvent on recovery of domoic acid : : (v:v:v) : (v:v) : (v:v) : (v:v) 8:1:1 7:2:1 6:3:1 5:4:1 1:1 9:1 17: (%) (%) (%)

4 1, : QuEChERS 超声时间的选择 DA, min,, 5 min, 4( 1.8 μg/g) 3.2 净化条件的选择 DA, QuEChERS PSA, C18, QuEChERS, [17] QuEChERS, 70%,,, 90%, 2, DA,, DA 3.3 方法检测低限线性范围回收率精密度 10 (S/N) DA 1.8 μg/g; 0.72~30.00 μg/ml, r μg/g 3 DA, QuEChERS, 6, HPLC, 3 Fig. 4 4 DA Effects of different ultrasonic time on recovery of domoic acid a: 5 min; b: 10 min; c: 15 min Table 2 表 2 基质匹配标准曲线法对 DA 回收率的影响 Effects of matrix-matched calibration standard on recovery of domoic acid (%) (%) (%)

5 76 6 Table 3 表 3 扇贝中 DA 加标回收率和精密度 (n=6) Recovery and precision of domoic acid spiking in scallops (n=6) (μg/g) (%) (%) (%) 结论 QuEChERS, DA, DA 20 μg/g,,, DA 参考文献 [1] Miles CO, Wilkins AL, Samdal IA, et al. A novel pectenotoxin, PTX-12, in Dinophysis spp. and shellfish from Norway [J]. Chem Res Toxicol, 2004, 17(11): [2] Fux E, McMillan D, Bire R, et al. Development of an ultraperformance liquid chromatography-mass spectrometry method for the detection of lipophilic marine toxins [J]. J Chromat A, 2007, 1157(1 2): [3] Stobo LA, Lacaze JPCL, Scott AC, et al. Liquid chromatography with mass spectrometry-detection of lipophilic shellfish toxins [J]. J AOAC Int, 2005, 88(5): [4] McNabb P, Selwood AI, Holland PT, et al. Multiresidue method for determination of algal toxins in shellfish: single-laboratory validation and interlaboratory study [J]. J AOAC Int, 2005, 88(3): [5]. (LC-MS/MS) [D]. :, Ma JG. Study on the simultaneous detection for multiple lipophilic toxins in shellfish using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) [D]. Qindao: Ocean University of China, [6] Intergovernmental Oceanographic Commission. Manual on harmful marine microalgae [R]. UNESCO, [7] Nguyen A, Luong J, Massona C. Capillary electrophoresis for detection and quantitation of domoic acid in mussels [J]. Anal Lett, 1990, 23(9): [8] Jennifer M. Maucher, John. Ultrasensitive detection of domoic acid in mouse blood by competitive ELISA using blood collection cards [J]. Toxicon, 2005, 45(5): [9] Dubois M, Demoulin L, Charlier C, et al. Development of ELISAs for detecting domoic acid, okadaic acid, and saxitoxin and their applicability for the detection of marine toxins in samples collected in Belgium [J]. Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2010, 27(6): [10] Vera-Avila LE, Deny Y, Marín-Pérez, et al. Trace level determination of domoic acid in seawater by off-line/on-line solid-phase extraction coupled to HPLC-UV [J]. J Mex Chem Soc, 2011, 55(2): [11] Maroulis M, Monemvasios I, Vardaka E, et al. Determination of domoic acid in mussels by HPLC with post-column derivatization using 4-chloro-7-nitrobenzo-2-oxa-1, 3-diazole (NBD-Cl) and fluorescence detection [J]. J Chromat B, 2008, 876(2): [12] McCarron P, Wright E, Quilliam AM. Liquid chromatography/ mass spectrometry of domoic acid and lipophilic shellfish toxins with selected reaction monitoring and optional confirmation by library searching of product ion spectra [J]. J AOAC Int, 2014, 97(2): [13] Furey A, Lehane M, Gillman M, et al. Determination of domoic acid in shellfish by liquid chromatography with electrospray ionization and multiple tandem mass spectrometry [J]. J Chromatogr A, 2001, 938(1 2): [14],,,. HPLC [J]. :, 2012, 31(3): Zhou XJ, Zhou XY, Zheng B, et al. HPLC method of rapid detecting domoic acid in shellfish [J]. J Zhejiang Ocean Univ: Nat Sci, 2012, 31(3): [15],,,. [J]., 2008, 27(S): Wu YX, Lin F, Lin HD, et al. Rapid determination of domoic

1, : QuEChERS 77 acid residues in amnesic shellfish by rapid resolution liquid 作者简介 chromatography [J]. Chin J Anal Lab, 2008, 27(S): 173 175.

Mod Food Sci Technol, 2011, 27(1): 120 122, 116. [17]. [J]., 2012, 22(5): 1 7. Zhu J.

6 1, : QuEChERS 77 acid residues in amnesic shellfish by rapid resolution liquid 作者简介 chromatography [J]. Chin J Anal Lab, 2008, 27(S): [16],,,. (DA) HPLC [J]., 2011, 27(1): , 116. Ji W, Zheng JY, Zeng XP, et al. HPLC analysis of domoic acid poisoning in south China sea [J]. Mod Food Sci Technol, 2011, 27(1): , 116. [17]. [J]., 2012, 22(5): 1 7. Zhu J. The new technology in analysis of harmful substances in food [J]. J Inspect Quarant, 2012, 22(5): 1 7. 王婵, 硕士, 主要研究方向为食品安全色谱检测 wangachan37@163.com 曹际娟, 研究员, 主要研究方向为食品安全检测 cjj0909@163.com ( 责任编辑 : 张宏梁 )

DOI /j.issn Food Research And Development UPLC-MS/MS 5 0.

DOI /j.issn Food Research And Development UPLC-MS/MS 5 0. 118 217 5 38 1 DOI 1.3969/j.issn.5-6521.217.1.28 Food Research And Development 1 2 1 2 1 2 1 2 1 2 1 2 1. 731 2. 731 - UPLC-MS/MS 5. 1 mol/l ProElut PXC 5 % - Thermo Hypersil Gold mm 2.1 mm 1.9 μm ESI

Microsoft Word 王 婵-校对 72 不要动版

Microsoft Word 王婵-校对 72 不要动版