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1 8 6 Vol. 8 No Journal of Food Safety and Quality Jun., 2017 C 聂 文 1, 屠泽慧 1, 张静 2, 蔡克周 1*, 姜绍通 1, 陈从贵 1 (1.,, ; 2., ) 摘要 : 目的 C 方法, 结果 C C 0.15%, 90% C 0.5 mg/ml, C 1, (1,1-diphenyl-2-picrylhydrazyl, DPPH) 90%; C 0.4 mg/ml, 84% 结论 C, C DPPH, 关键词 : ; C; ; Effects of vanillin and vitamin C on the content of heterocyclic amines in braised beef NIE Wen 1, TU Ze-Hui 1, ZHANG Jing 2, CAI Ke-Zhou 1*, JIANG Shao-Tong 1, CHEN Cong-Gui 1 (1. Key Laboratory for Agricultural Products Processing of Anhui Province, School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei , China; 2. Anhui Huaibei Productivity Promotion Center, Huaibei , China) ABSTRACT: Objective To explore the effects of vanillin and vitamin C on the content of heterocyclic amines during the process of beef stewing. Methods The solid phase extraction pretreatment technique was used to determine the content of heterocyclic amines in meat samples under high phase liquid phase ultraviolet rays and fluorescence detection. Result Vanillin and vitamin C were able to reduce the formation of heterocyclic amines in braised beef. When the concentration of vitamin C added to 0.15%, the total amount of heterocyclic amines decreased by 90% or more. When the vitamin C concentration increased to 0.5 mg/ml, vitamin C on DPPH free radical scavenging rate was up to 90%, and when added vitamin C concentration was 0.4 mg/ml, the superoxide anion clearance rate was up to 84%. Conclusion Vitamin C and vanillin can reduce the formation of heterocyclic amines in braised beef. The scavenging rate of DPPH free radicals and superoxide anion radicals of vitamin C are higher than those of vanillin, and the inhibitory effect on heterocyclic amines is better than vanillin. KEY WORDS: vanillin; vitamin C; braised beef; heterocyclic amines 基金项目 : ( ) Fund: Supported by National Natural Science Foundation of China ( ) * 通讯作者 :,,, kzcai@hfut.edu.cn *Corresponding author: CAI Ke-Zhou, Associate Professor, College of Food Science and Engineering, Hefei University of Technology, No. 193, Tunxi Road, Hefei , China. kzcai@hfut.edu.cn

2 引言,, (Maillard) [1,2], [3,4] 30 [5] 30 (aminoimidazo azaarens, AlAs) 2, / 300 [6] ; 300 [7] 30,, [1,8],, [9,10] 1, (1,1-diphenyl-2-picrylhydrazyl, DPPH) [11,12] DPPH, Maillard [13],, Wong [14] B 2 (PhIp MeIQx) VB 2 VB 3 VB 5 VB 9 VB % Cheng [15] 12 :, (butyl hydroxyanisole, BHA) (butylated hydroxytoluene, BHT), BHT [16] [17] E [18] [19] [20], C, 0.05% 0.10% 0.15% C,, DPPH 2 材料与方法 2.1 材料与试剂 ( ) (, ); (, ); ( ) : [4,5-b] (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, PHIP) (CAS: , : ); H- [4,3-b] (3-amino-1-methyl-5H-pyrido[4,3-b]indole, Trp-P-2)(CAS: , : ); 2- -3,4,8- -3H- [4,5-f] (2-amino-3,4,8-trimethyl- 3H-imidazo[4,5-f]quinoxaline, 4,8-DimeIQX)(CAS: , : ); 3- -1,4- -5H- [4,3-b] (3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole, Trp-P-1) (CAS: , : ); 2- -3,4- -3H- [4,5-f] (2-amino-3,4-dimethyl-3H-imidazo [4,5-f]quinoxaline, MeIQ)(CAS: , : ); 2- -3,8- [4,5-f] (2-amino-3,8- dimethylimidazo[4,5-f]quinoxaline, MeIQX)(CAS: , : ); 1- -9H- [3,4-b] (1-methy-9H-pyrido[3,4-b]indole, Harman)(CAS: , : ); 2- -9H- [3,4-b] (2-amino-9H-pyrido[2,3-b]indole, AαC)(CAS: , : ); H- [4,5-f] (2-amino-3-methyl-3H-imidazo[4,5-f]quinoxaline, IQ)(CAS: , : ) 99%, TRC 2.2 仪器与设备 G1315A G1312 Agilent1100 Bond Elut-PRS (500 mg/3 ml) Bond Elut-C 18 (500 mg/3 ml)( Agilent ); AR1140 ( ( ) ); BCD-249CF ( ); DGF30/7-Ⅰ ( ); SHZ-D( Ⅲ) ( ); CR15RT ( ); T 6 ( );

3 6, : C 1989 ASE-12 ( ); HH-2 ( ); GW-4 ( ); ZSBN-3004 ( 犇 ) 2.3 原料前处理, 5 mm 1 cm 5 cm, 50 g : 1 g 1 g 5 ml 150 ml 250 ml ( 98 ) 100 g,, 2 h, 30 min 1 ( ),, -18 : C : 1 g 1 g 5 ml 150 ml 250 ml ( 98 ) 50 g g(0.05%) 0.05 g(0.1%) g(0.15%) C,, 2 h, 30 min 1 ( ), : 1 g 1 g 5 ml 150 ml 250 ml ( 98 ) 50 g g(0.05 %) 0.05 g(0.1 %) g(0.15 %),, 2 h, 30 min 1 ( ), : 1 g 1 g 5 ml 150 ml 250 ml ( 98 ) 50 g,, 2 h, 30 min ( ), : 1 g 1 g 5 ml 150 ml 250 ml 50 g, 2 h 2.4 杂环胺检测 标准曲线的制作 9 1 mg 10 ml 100 μg/ml, ng/ml, 杂环胺的提取 2 g 50 ml, 12 ml 2 mol/l NaOH, r/min 20 min 10 g,, (propyl sulfonic acid, PRS), PRS 5 ml,, 70 ml, 2 /3 s 高效液相色谱条件 (MeIQX MeIQ IQ DimeIQX), 63 nm; (Trp-P-2 Harman Trp-P-1 AαC) PHIP, 300 nm, 440 nm TSK-Gel ODS-80TM(250 mm 4.6 mm, 5 μm); 30 ;, A: ; B: 0.05 mol/l - (ph=3.6), 1 Table 1 表 1 流动相梯度洗脱顺序 Gradient elution program of mobile phase /min A( ) B( - ) 0~25 90% 10% 25~35 50% 50% 35 90% 10% 2.5 DPPH 清除率的测定 mg DPPH 10 ml, DPPH 0.5 mmol/l, (0 4 ); 4 ml 0.1 mmol/l DPPH 4 ml ( mg/ml) C,, 1 h, 515 nm A i 4 ml, 515 nm A 0 4 ml 4 ml 515 nm A j, 3 DPPH A0 Ai Aj DPPH 自由基清除 /% 100 A 2.6 超氧阴离子清除率 3 ml 0.05 mol/l Tris-HCl (ph=8.2), min, ( mg/ml), 0.6 ml 30 mmol/l, min, 0.5 mol/l 1 mol/l HCl, 325 nm A 1 ; 325 nm, 0

4 A 2 A 3 = 1 ( A1 A2) 100% A 3, 3 结果与分析 3.1 杂环胺标准曲线 杂环胺含量的结果分析 3 4, 1 : C (P<0.05), C,, C 0.15%, 99%, C 表 2 9 种杂环胺的线性范围 线性方程 相关系数和检出限 Table 2 Linear ranges, linear equations, correlation coefficients and detection limits of 9 heterocyclic amines (ng/ml) (ng/ml) DimeIQx 5~500 Y=0.3095X MeIQx 5~500 Y=0.3891X IQ 5~500 Y=0.38X Harman 5~500 Y=0.2645X Trp-P-1 5~500 Y=0.2607X Trp-P-2 5~500 Y=0.3274X AαC 10~500 Y=0.1863X PHIP 10~500 Y=0.13X MeIQ 10~500 Y=0.2069X : Y ; X, ng/ml Table 3 表 3 不同浓度不同添加物对牛肉极性杂环胺含量的影响 (ng/g, n=3) Effects of different concentrations and different additives on the content of polar heterocyclic amine in beef (ng/g, n=3) IQ DimeIQX MeIQX PHIP MeIQ ND ND ND 0.16±0.01 e ND 76.08±1.57 a 12.57±1.03 a 2.77±0.02 a 3.37±0.22 a 2.64±0.31 a 0.05 % ND 0.97±0.03 e 0.13±0.01 d 0.19±0.01 e 0.38±0.06 e 0.1 % 1.69±0.13 b 2.99±0.02 c 0.65±0.01 c 0.60±0.06 c 0.98±0.03 d 0.15% 3.71±0.15 c 4.21±0.11 b 1.18±0.02 b 1.25±0.07 b 1.40±0.17 c C 0.05% ND 1.81±0.05 d 0.63±0.04 c 0.27±0.01 d 1.77±0.13 b C 0.1% ND 0.57±0.03 e ND 0.16±0.01 e 1.03±0.10 d C 0.15% ND ND ND ND 0.22±0.01 e : ND ; (P<0.05) 表 4 不同浓度不同添加物对牛肉非极性杂环胺含量的影响 (ng/g, n=3) Table 4 Effect of different concentrations and different additives on the content of nonpolar heterocyclic amine in beef (ng/g, n=3) Trp-P-2 Harman Trp-P-1 AαC ND ND ND ND 3.71±0.03 a ND 2.85±0.04 a 6.49±0.31 a 0.05% 1.66±0.12 b ND 1.23±0.11 b 1.67±0.02 b 0.1% 0.78±0.03 c ND ND 0.45±0.02 c 0.15% ND ND ND ND C0.05% 0.57±0.02 c 2.46±0.21 a 0.32±0.02 c 1.53±0.03 b C0.1% ND 1.88±0.14 b 0.30±0.01 c 0.42±0.01 c C0.15% ND ND 0.17±0.01 d ND : ND ; (P<0.05)

5 6, : C 1991 C 0.4 mg/ml,, 84%, 40%, C : C, Fig. 1 1 (n=3) Effect of additives on total amount of heterocyclic amines (n=3) 3.3 DPPH 清除率结果分析 DPPH 0.8 mg/ml, mg/ml 2 DPPH (P<0.05), C DPPH, 0.5 mg/ml, DPPH 90%, DPPH 30% : C DPPH, 2 DPPH (n=3) Fig. 2 Determination of DPPH free radical scavenging rate (n=3) 3.4 超氧阴离子清除率结果分析 3,, C, (P<0.05) C 0.4 mg/ml, 3 (n=3) Fig. 3 Determination of superoxide anion clearance rate (n=3) 4 结论 C, C, (P<0.05), C 0.15%, 90% C DPPH ; 0.5 mg/ml, C DPPH 90%, DPPH 30% ; C 0.4 mg/ml,, 84%, 40% C, C DPPH,, 参考文献 [1],,,. [J]., 2010, 31(23): Yao Y, Peng ZQ, Shao B, et al. Heterocyclic aromatic amines in processed meat products [J]. Food Sci, 2010, 31(23): [2],,,. [J]., 2017, 8(3): Yang TT, He ZY, Qin F, et al. Research progress of the effects of Maillard reaction on flavor and quality of products as well as derivatized harmful substances [J]. J Food Saf Qual, 2017, 8(3):

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