Microsoft Word 朱珍-二校

Size: px

Start display at page:

Download "Microsoft Word 朱珍-二校"

衡范
9 years ago
Views:

1 6 4 Vol. 6 No Journal of Food Safety and Quality Apr., 2015 朱珍 1, 朱俊霖 1, 庄松娟 2, 李振兴 1* (1., ; 2., ) 摘要 : 目的方法 0.1%,,,,,,, Aglient ZORBAX Eclipse-C 18, -0.1%, 275 nm, 305 nm, 0.8 ml/min 结果 µg/kg 72.20%~85.68%, (RSD) 1.716%~9.926% 结论, > > 关键词 : ; ; ; Detection of dissociative o-tyrosine in electron beam irradiated seafood by high performance liquid chromatography ZHU Zhen 1, ZHU Jun-Lin 1, ZHUANG Song-Juan 2, LI Zhen-Xing 1* (1.Food Safety Laboratory, Ocean University of China, Qingdao , China; 2. Qingdao Institute for Food and Drug Control, Qingdao , China ) ABSTRACT: Objective To establish a method for the determination of o-tyrosine in irradiated seafood by high performance liquid chromatography (HPLC). Methods The sample was homogenized with 0.1% formic acid, the ultrasonic bath and centrifugation were followed. Then the resulting supernatants were treated with acetone at 20 for 2 h and centrifuged again. After evaporation of acetone from supernatants, the remaining solution was dried freeze. Finally, the powder was collected, reconstituted in formic acid and detected by HPLC analysis with external reference method. For HPLC separation, Aglient ZORBAX Eclipse-C 18 column was used with a flow rate at 0.8 ml/min, mobile phase: methanol-0.1% formic acid in Milli-Q water, and the fluorescence detector was set at λex275 nm and λem305 nm. Results The recoveries of o-tyrosine at the spiked amounts of 50, 200, and 500 μg/kg were 72.20%~85.68% with RSD value between 1.716% to 9.926%. Conclusion This method can be applied to detect o-tyrosine in electron beam irradiated seafood. The content of dissociative o-tyrosine in some seafood was metapenaeusensis>lateolabrax japonicas> scallop. KEY WORDS: electron beam irradiation; o-tyrosine; high performance liquid chromatography; seafood 基金项目 : (2013GGB01192) Fund: Supported by Science and Technology Development Plan Project of Shandong Province (2013GGB01192) * 通讯作者 :,,, [email protected] *Corresponding author: LI Zhen-Xing, Associate Professor, Food Science and Engineering College, No.5, Yushan Road, Qingdao , China. [email protected]

926% 结论, > > 关键词 : ; ; ; Detection of dissociative o-tyrosine in electron beam irradiated seafood by high performance liquid chromatography ZHU Zhen 1, ZHU Jun-Lin 1, ZHUANG Song-Juan 2, LI

2 4, : 引言,, 60 Co 137 Cs gamma 5MeV X [1], [2,3] [4], [5],, [6], 39.5%, DNA [7,8], gamma,,, - ( ),, [9],, : ( ) ( ),, [10,11],,, 2 材料与方法 2.1 材料与试剂 DL- ( 98%, Sigma ); (, TEDIA ); (, );, (, ); Milli-Q ; (1 ml); ( 66, 0.22 μm); (2 ml,,,, Agilent ) ( ) 2.2 实验仪器与设备 (Agilent 1260, Aglient ); (Hei-Vap, Heidolph ); (DS-1, ); (3K-15, Sigma ); (KQ5200B, ); ( 0.1 g, TE601-2, ); ( g, BS-224-S, ); (Milli-Q, Millipore ); Aglient ZORBAX Eclipse-C 18 (4.6 mm 250 mm, 5.0 µm, Aglient ) 2.3 标准溶液的配制 : 5 mg, 0.1% 5 ml, 4 : 0.1%, 样品处理游离邻酪氨酸的提取 Chen [10], 0.8 g ( 0.01 g) 50 ml, 3.5 ml 0.1%, r/min 2 min, 15 min, 8000 r/min, 4 20 min,, 8 ml, h, 8000 r/min, 4 15 min, 45,, 0.8 ml 0.1%, 0.22 µm, 结合邻酪氨酸的提取 [11], g ( g) 50 ml, 20 ml - (3:1),, 8000 r/min, 4 10 min,,,,,, 6 ml, 6 mol/l, 110,,

2 实验仪器与设备 (Agilent 1260, Aglient ); (Hei-Vap, Heidolph ); (DS-1, ); (3K-15, Sigma ); (KQ5200B, ); ( 0.1 g, TE601-2, ); ( 0.

3 色谱条件及参数非衍生高效液相色谱 : AglientZORBAXEclipse-C 18 (4.6 mm 250 mm, 5.0 µm); : 0.1% (A)- (B); : 0~14 min, 92%(A); 14~14.2 min, 92%(A)~20%(A); 14.2~21 min, 20%(A); 21~21.2 min, 20%(A)~92%(A); 21.2~30 min, 92%(A); : 20 µl; : 30 ; 275 nm, 305 nm 柱前衍生 - 高效液相色谱 OPA : g, g, g, 50% 10 ml, 0.22 µm, 4 : =1:40 (v:v), 1 min 20 µl : AglientZORBAXEclipse-C 18 (4.6 mm 250 mm, 5.0 µm); : 0.02 mol/l (A)- (B); : 0~10 min, 80%~65%(A), 10~15 min, 65%(A), 15~15.01 min,65%~80%(a), 15.01~20 min, 80%(A); : 30 ; 330 nm, 450 nm 3 结果与分析 3.1 方法的线性范围检出限定量限精密度 -, 0.1~9500 µg/l, 0.8 µg/l, 5 µg/l, ( 1),,,,, : µg/l, (Y) (X, µg/l), 1~10000 µg/l, Y=0.0503X , , 2 (S/N=3) 2.7 µg/l, (S/N=10) 8 µg/l A: OPA - A: Chromatogram of o-tyrosine standards by pre-column derivatization HPLC with OPA B: B: Chromatogram of o-tyrosine standards by HPLC without derivatization Y: Y: orth o-tyrosine 1 Fig. 1 Liquid chromatogram of o-tyrosine standards

22 µm, 4 : =1:40 (v:v), 1 min 20 µl : AglientZORBAXEclipse-C 18 (4.6 mm 250 mm, 5.0 µm); : 0.02 mol/l (A)- (B); : 0~10 min, 80%~65%(A), 10~15 min, 65%(A), 15~15.01 min,65%~80%(a), 15.

4 4, : Fig. 2 Standard curve of o-tyrosine, 6, 1 1,,, (RSD) 10% 3.2 方法的加标回收率, µg/kg, 3, , 2, 72.20%~85.68%, 1.716%~9.926% 3.3 辐照样品的测定 ( 10 kw, 10 MeV) 10 kgy ,, 3: 表 1 鲈鱼基围虾扇贝中精密度实验结果 (n=6) Table 1 Results of precision test (n=6) o-tyrosine /ng Average/ng RSD/% Lateolabraxjaponicus Metapenaeusensis Scallop 表 2 鲈鱼基围虾扇贝中邻酪氨酸的加标回收率和相对标准偏差 (n=3) Table 2 Recoveries and RSDs of o-tyrosine in real samples by LC-FLD (n=3) (%) (%) (%) (%) (%) (%) HPLC-FLD 50 µg/kg 200 µg/kg 500 µg/kg Lateolabraxjaponicus Metapenaeusensis Scallop 表 3 Table 3 辐照水产品中的邻酪氨酸含量 (n=3) Content of o-tyrosine in seafood (n=3) µg/kg µg/kg Lateolabraxjaponicus Metapenaeusensis Scallop : 10 kgy,

5,, 3: 表 1 鲈鱼基围虾扇贝中精密度实验结果 (n=6) Table 1 Results of precision test (n=6) o-tyrosine /ng 1 2 3 4 5 6 Average/ng RSD/% Lateolabraxjaponicus 114.589 106.731 116.554 114.589 112.625 114.

5 1450 6,, 10 kgy 4 讨论 4.1 实验原理初探 : -, OPA( ), [11] ; [12],,,,, :, 110, HPLC [13] ;,,,,,, HPLC [14] -,,, ;,, [8], 4.2 实验结果讨论, -,,, ;, 80%, Chen [10] µg/kg,,, [15], mg/100 g, 10-6 ~10-5 [16] HPLC-MS-MS 10 kgy, mg/kg,,, ; [8] 10 kgy, 178.5/288 µg/kg,,, ; Wolfgang [17] kgy,, 10 kgy µg/g, 5 结论与展望,,,,, > >,,,,, 参考文献 [1]. [J]., 2013, (15): Geng JN. Food irradiation and its application in food processing [J]. Food Res Devel, 2013, (15): [2],,,. [J]., 2007, (01): Ha YM, Shi HD, Wang F, et al. Application and characteristics of food irradiation by electron beam [J]. J Nucl Agric Sci, 2007, (01): [3] Peter B, Roberts. Food irradiation is safe:half a century of studies[j].radiat Phys Chem, 2014, (105): [4],,,.

5/288 µg/kg,,, ; Wolfgang [17] 0 2 4 6 8 10 kgy,, 10 kgy 45 17 8 3 µg/g, 5 结论与展望,,,,, > >,,,,, 参考文献 [1]. [J]., 2013, (15): 109 112. Geng JN.

6 4, : 1451 [J]., 2012, 2(4): Li ZX, Zhang LM, Gu KF, et al.comparative study and suggestion on relevant standards and regulations fordomestic and international aquatic irradiation [J].Chin Fish Qual Stand, 2012, 2(4): [5],,. [J]., 2010, 4: Yu L, Bu Y, Wei ZP. The application study of irradiation on aquatic products [J]. Meat Res, 2010, 4: [6]. [M]. :, 2014 National Bureau of Statistics of the People's Republic of China. China statistical yearbook [M]. Beijing: Chinese Statistics Press, [7],,,. [J]., 2009, (5): Wang F, Wang ZD, Zhou HJ, et al. The methods and standards of detection techniques for irradiated foods [J]. Food Mech, 2009, (5): [8],. [J]., 2005, (06): Ha YM, Zhou HJ. The reviewon analytical detection methodsfor irradiated foods [J]. Food Sci, 2005, (06): [9],,,, [J]., 2011, 30(10): Wu M, Li JX, Zhang ZG, et al. Identification of irradiated protein-rich food by dissociative tyrosine [J].J Instrum Anal. 2011, 30(10): [10] Chen S, Fan LH, Song J, et al. Identification of irradiated meats by determining o- and m-tyrosine as markers [J]. Meat Sci, 2013, 93(2): [11],,,. γ- Lin L, Zhang K, Qian H. Determination of o-tyrosine in irradiated chicken products by HPLC [J]. Food Sci, 2010, (10): [13],,,. [J]., 2011, (01): Zheng R, Ding ZP, Yan LM, et al. Study pre-treatment of HPLC determinatethe o-tyrosine in irradiation food [J].Shanghai Meas Test, 2011, (01): [14] Ibe FI, Grinter R, Massey R, et al. Detection of o-tyrosine in irradiated chicken by reverse-phase HPLC and fluorescence detection [J]. Food Add Contam, 1991, (8): [15], ( ) [M]. :, Wang YD, China Food Composition (The Second Edition) [M].Beijing: Peking University Press,2009. [16],,,. HPLC-MS/MS [J]., 2011, 30(10): Wang M, Zhang XQ, Yang ZY, et al. Determination of o-tyrosine in γ-ray irradiation protein-rich food by HPLC-MS/MS [J]. J Instrum Anal, 2011, 30(10): [17] Wolfgang G, Hein TJ, Simat HS. Determination of non-protein bound o-tyrosine as a marker for the detection of irradiated shrimps [J]. Eur Food Res Technol, 2000, (210): 作者简介 ( 责任编辑 : 李振飞 ) 朱珍, 硕士研究生, 主要研究方向为辐照食品的检测 @139.com [J]., 2009, (10): Wang DH, Wang M, Han L, et al.determination of o-tyrosine in γ-ray irradiated food by HPLC [J]. Food Sci, 2009, (10): [12],,. 李振兴, 博士, 副教授, 主要研究方向为水产品的安全与质量控制 [email protected] [J]., 2010, (10):