Microsoft Word 刘功良.doc

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1 5 6 Vol. 5 No Journal of Food Safety and Quality Jun., 2014 刘功良, 赵晓娟, 陈艺琦, 陈海光 *, 白卫东 (, ) 摘要 : 目的方法,, ph ph, 结果, : 0.15 U/L min 3.5 h, Y=0.024X-0.013(, Y ΔpH, X ), R 2 = , IC μg/l, 0~50 μg/l, %~124.58%, 4.39%~16.46% 结论关键词 : ; ; ; Research on microbial inhibition method for rapid detection of erythromycin in raw milk LIU Gong-Liang, ZHAO Xiao-Juan, CHEN Yi-Qi, CHEN Hai-Guang *, BAI Wei-Dong (College of Light Industry and Food Technology, Zhongkai University of Agriculture and Technology, Guangzhou , China) ABSTRACT: Objective To determine the concentration of erythromycin in raw milk by microbial inhibition method. Methods After sterilized and cooled the raw milk, erythromycin was added into raw milk samples with Lactobacillus, and the blank control was set. After fermented for some time, the ph of the fresh milk samples before and after fermentation was detected for testing the difference of ph of raw milk samples while the blank control was set. Results After single factor and orthogonal experiment, the optimum experimental conditions for Lactobacillus fermentation were established as follows: inoculum concentration of Lactobacillus CH-1 was 0.15 U/L, fermentation temperature was 40, fermentation time was 3.5 h and the dissolution time of CH-1 lactobacillus powder was 25 min. According to the relationship between doxycycline concentration and difference of ph value, the standard curve was established. The standard curve equation was as follows: Y=0.024X (Y was the concentration of erythromycin in raw milk samples, X was the ΔpH). The linear 基金项目 : (201104); (S ); (2012KJCX0068) Fund: Supported by the Open Project Program of Provincial Key Laboratory of Green Processing Technology and Product Safety of Natural Products in Guangdong Province(201104), Natural Science Foundation of Guangdong Province(S )and Educational Commission of Guangdong Province(2012KJCX0068) * 通讯作者 :,, chg1508@163.com *Corresponding author: CHEN Hai-Guang, Professor, College of Light Industry and Food Technology, Zhongkai University of Agriculture and Technology, No.501, Zhongkai Road, Haizhu District, Guangzhou, , China. chg1508@163.com

2 correlation coefficient R 2 was The detection range of the standard cure was 0~50 μg/l. The microbial inhibition method showed that the sensitivity IC 50 was μg/l for erythromycin. The assay showed a mean recovery ranged from %~124.58%, and the coefficients of variations for the inter-assay was 4.39%~16.46%. Conclusion It could be used to detect the erythromycin residual concentration in raw milk. KEY WORDS: raw milk; lactic acid bacteria; erythromycin; fast detection 1 引言,, [1-3],,, [4-6],,, [7-9],,, [10-13],, [14-16],, 2 材料与方法 CH-1,, ( 40 μg/l) ;, ph ph, ph (ph 1 ) ph (ph 2 ), ΔpH(ΔpH= ph 2 ph 1 ) 4 ΔpH IBM SPSS Statistics,, ΔpH ΔpH-, 3 结果与分析 3.1 乳酸菌发酵产酸的优化 0.2 U/L CH-1,, 40 μg/l, 45, 0.5 h ph 1, 1 h, ph, CH-1 2, CH U/L, 2.1 实验材料试剂与主要仪器 ;, 850 μg/mg; CH-1 ; PHS-25 ph 2.2 实验方法 (120, 30 min),, 1 CH-1 Fig. 1 Effect on fermentation and acid production of Lactobacillus CH-1 by erythromycin

3 6, : 1753 Fig. 2 2 ph Effect of Lactobacillus concentration on the difference of ph value Fig. 4 4 ΔpH Effect of fermentation time on the difference of ph value ΔpH, ΔpH 0.15 U/L ; 0.15 U/L 0.2 U/L, ΔpH 3 h ; CH U/L, ΔpH 2.5 h, 0.15 U/L 0.2 U/L, CH U/L 3, 55 60, ΔpH,,, CH , ΔpH 3.5 h, 35, ΔpH 3.5 h,, CH-1, CH-1, CH , ΔpH, 3.5 h,, CH h 5, ΔpH, 20 min, 3.5 h ΔpH, CH-1 20 min Fig. 5 5 ph Effect of dissolution time of lactic acid bacteria powder on the difference of ph value,, 1, L 9 (3 4 ), ΔpH, 2 2, : > >>, A 2 B 1 C 2 D 3, 0.15 U/L min 3.5 h, ph ph Fig. 3 3 ΔpH Effect of fermentation temperature on the difference of ph value 表 1 四因素三水平正交实验 Table 1 Orthogonal layout L 9 (3 4 ) A B C D U/L / /h /min

4 Table 2 表 2 正交实验结果与分析 Experiment result calculation and analysis of orthogonal layout A U/L B / C /h D /h ΔpH K K K k k k R 检测标准曲线的建立 6, ΔpH, 1~50 μg/l ΔpH 50 μg/l, ΔpH 5, 0~50 μg/l, ΔpH,, Y=0.024X 0.013, 7 Fig. 7 Standard curve of erythromycin detected with microbial inhibition method R , IC μg/l, IC μg/l 3.3 回收率实验 3, 15 μg/l 30 μg/l 45 μg/l,,, %~103.54%, 9.80%~6.26% 3.4 重复性实验 15 μg/l 30 μg/l 45 μg/l,,, 4, 4.39%~16.46% 表 3 微生物抑制法检测红霉素的回收率 Table 3 Recovery range of erythromycin detected with microbial inhibition method (%, n=3) ± ± ± 表 4 微生物抑制法检测红霉素的同一批次间精确度分析 Table 4 Accuracy analysis of detection erythromycin in the same interassay detected with microbial inhibition method (CV, %) Fig ~100 μg/l ΔpH Effect of erythromycin (0~100 μg/l) on the difference of ph value ± ± ±

5 6, : 总结与展望 4 h, TTC 20 [14,15],,,,,,, 参考文献 [1] Frasconi M, Tel-Vered R, Riskin M, et al. Surface Plasmon resonance analysis of antibiotics using imprinted boronic acidfunctionalized Au nanoparticle composites [J]. Anal Chem, 2010, 82 (6): [2],,. [J]., 2011, 45(7): Wang JJ, Gao MZ, Bao JM. Progress on the application of liquid chromatography- tandem mass spectrometry in examination of animal residue [J]. Chin J Veterinary Drug, 2011, 45(7): [3] Zhang J, Zhang B, Wu Y, et al. Fast determination of the tetracyclines in milk samples by the aptamer biosensor[j]. Analyst, 2010, 135(10): [4],,,. [J]., 2012, 40(9): Chen Y, Li HL, Zhang DJ, et al. Study on the residues and detected methods of antibiotics in milk [J]. China Dairy Ind, 2012, 40(9): [5] Spier CR, Vadas GG, Kaattari SL, et al. Near real-time, on-site, quantitative analysis of PAHs in the aqueous environment using an antibody-based biosensor [J]. Environ Toxicol Chem/SETAC, 2011, 30(7): [6] Li YB, Zhang ZJ, Li JS, et al. Simple, stable and sensitive electrogenerated chemiluminescence detector for high-performance liquid chromatography and its application in direct determination of multiple fluoroquinolone residues in milk [J]. Talanta, 2011, 84(3): [7] Mor F, Kocasari FS, Ozdemir G, et al. Determination of sulphonamide residues in cattle meats by the Charm-II system and validation with high performance liquid chromatography with fluorescence detection [J]. Food Chem, 2012, 134: [8] Wang W, Zhang Y, Wang J, et al. Determination of β-agonists in pig feed, pig urine and pig liver using capillary electrophoresis with electrochemical detection [J]. Meat Sci, 2010, 85: [9],. [J]., 2013, 4(1): Gao J, Miao H. Recent development of analytical methods for veterinary drug residues [J]. J Food Safe Qual, 2013, 4(1): [10],,. [J]., 2010, 35(6): Wang LP, Cai XF, Hou CY. Study on microbial inhibition assay of sulbactam in milk [J]. Food Sci Technol, 2010, 35(6): [11],,,. [J]., 2008, 18(9): Wang ZQ, Hu GY, Li ZY, el al. Rapid determination of antibiotics residues in animal derived food by microbial inhibition method [J]. Chin J Health Lab Technol, 2008, 18(9): [12] Pikkemaat MG, Rapallini ML, Karp MT, et al. Application of a luminescent bacterial biosensor for the detection of tetracyclines in routine analysis of poultry muscle samples [J]. Food Add Contam Part A, 2010, 27(8): [13]. [J]., 2010, 33(4): Chu LQ. Study on antibiotic residues in milk and its comparative analysis of three kinds of detection methods [J]. J Dairy Sci Technol, 2010, 33(4): [14],,. [J]., 2010, 49(6): Liu JA, Gao MH, Huang YL. Advances and prospects of detecting techniques for antibiotic residues in milk [J]. Hubei Agric Sci, 2010, 49(6): [15],,,. [J]., 2010, 31(8): Yu Y, Hu XL, Chao GQ, et al. Emulsified rapid detection of antibiotic residues method of inquiry [J]. Food Res Dev, 2010, 31(8): [16],,. [J]., 2010, 38(11): Li XH, Wang FF, Chi T. Study of technology for antibiotic residues in milk and the detection method [J]. China Dairy Ind, 2010, 38(11): ( 责任编辑 : 邓伟 )

1756 5 作者简介刘功良, 博士, 副教授, 硕导, 主要研究方向为食品安全检测 E-mail:

com 陈海光, 教授, 硕导, 主要研究方向为食品加工与安全 E-mail: chg1508@163.

6 作者简介刘功良, 博士, 副教授, 硕导, 主要研究方向为食品安全检测 gongliangliu@126.com 陈海光, 教授, 硕导, 主要研究方向为食品加工与安全 chg1508@163.com 食品贮藏与保鲜专题征稿投稿方式 : tougao@chinafoodj.com 食品安全质量检测学报编辑部

Microsoft Word - 3098-3102 20150611007-蔡伟江-二校

Microsoft Word - 3098-3102 20150611007-蔡伟江-二校 6 8 Vol. 6 No. 8 2015 8 Journal of Food Safety and Quality Aug., 2015 蔡伟江 *, 邓鹏飞, 刘光兰, 李丹 (, 519040) 摘要 : 目的, 方法 (4.6 mm 250 mm, 5.0 μm) ; : : : =70: 16: 14(V:V:V),, : 1.0 ml/min; : 40 ; : 10