Full Text:   <1750>

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CLC number: TS252.7

On-line Access: 2015-08-04

Received: 2014-12-20

Revision Accepted: 2015-05-11

Crosschecked: 2015-07-09

Cited: 0

Clicked: 3476

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Zhan-mei Jiang

http://orcid.org/0000-0002-7296-658X

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Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.8 P.672-679

http://doi.org/10.1631/jzus.B1400359


Development of a test strip for rapid detection of lactoperoxidase in raw milk


Author(s):  Hong-xia Che, Bo Tian, Li-na Bai, Li-ming Cheng, Li-li Liu, Xiao-na Zhang, Zhan-mei Jiang, Xiao-xi Xu

Affiliation(s):  Key Laboratory of Dairy Science (KLDS), Ministry of Education, Northeast Agricultural University, Harbin 150030, China

Corresponding email(s):   zhanmeijiang@neau.edu.cn, xiaoxi_xu01@163.com

Key Words:  Lactoperoxidase, Test strip, Rapid detection, Raw milk, 3, 3'5, 5'-Tetramethylbenzidine (TMB)


Hong-xia Che, Bo Tian, Li-na Bai, Li-ming Cheng, Li-li Liu, Xiao-na Zhang, Zhan-mei Jiang, Xiao-xi Xu. Development of a test strip for rapid detection of lactoperoxidase in raw milk[J]. Journal of Zhejiang University Science B, 2015, 16(8): 672-679.

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author="Hong-xia Che, Bo Tian, Li-na Bai, Li-ming Cheng, Li-li Liu, Xiao-na Zhang, Zhan-mei Jiang, Xiao-xi Xu",
journal="Journal of Zhejiang University Science B",
volume="16",
number="8",
pages="672-679",
year="2015",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1400359"
}

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%T Development of a test strip for rapid detection of lactoperoxidase in raw milk
%A Hong-xia Che
%A Bo Tian
%A Li-na Bai
%A Li-ming Cheng
%A Li-li Liu
%A Xiao-na Zhang
%A Zhan-mei Jiang
%A Xiao-xi Xu
%J Journal of Zhejiang University SCIENCE B
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1400359

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T1 - Development of a test strip for rapid detection of lactoperoxidase in raw milk
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A1 - Bo Tian
A1 - Li-na Bai
A1 - Li-ming Cheng
A1 - Li-li Liu
A1 - Xiao-na Zhang
A1 - Zhan-mei Jiang
A1 - Xiao-xi Xu
J0 - Journal of Zhejiang University Science B
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SP - 672
EP - 679
%@ 1673-1581
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1400359


Abstract: 
Traditional methods for detecting lactoperoxidase (LP) are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid (CA)/0.2 mol/L disodium hydrogen phosphate (NaP) buffer solution (pH 5.0), 22 mmol/L 3,3',5,5'-Tetramethylbenzidine (TMB), 0.6 mmol/L hydrogen peroxide (H2O2), and 0.5% Tween-20 or 0.3% cetyltrimethyl ammonium bromide (CTAB) were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation (CV) of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1ȁ3;2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation (r=0.9776). So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.

乳过氧化物酶快速检测试纸的研制

目的:开发快速、灵敏和准确的乳过氧化物酶检测方法,用于原料乳验收及相关乳制品加工中对乳过氧化物酶的检测。
创新点:首次以试纸检测方法代替传统的试管检测法,借助便携式光电测色仪实现对乳过氧化物酶的定量检测分析。
方法:以反光值为检测指标,从缓冲液pH、四甲基联苯胺(TMB)浓度、过氧化氢(H2O2)浓度和表面活性剂四个方面,优化筛选检测试纸的最佳制作条件;通过精密度试验、最低检出浓度和与试管检测法相关性分析,评价试纸检测法的可靠性和准确性。
结论:试纸检测体系的优化结果为0.1 mol/L柠檬酸/0.2 mol/L磷酸氢二钠溶液(pH 5.0),22 mmol/L TMB,0.5% Tween-20或0.3% 溴化十六烷基三甲铵(CTAB),和0.6 mmol/L H2O2。检测试纸的精密度较好,变异系数在 2.47%~6.72%(表1);最低检出浓度为1.000~2.000 mg/L(图7);与试管法检测结果的相关系数为0.9776(图8)。因此,试纸检测法是一种快速、灵敏和准确的乳过氧化物酶检测技术,可满足原料乳验收及相关乳制品加工中对乳过氧化物酶检测的需求。

关键词:乳过氧化物酶;试纸;快速检测;原乳;四甲基联苯胺

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]Bardsley, W.G., 1985. Steady-state kinetics of lactoperoxidasecatalyzed reaction. In: Pruitt, K.M., Tenovuo, J.O. (Eds.), The Lactoperoxidase System: Chemistry and Biological Significance. Marcel Dekker, New York, p.55-87.

[2]Boulares, M., Mankai, M., Hassouna, M., 2011. Effect of activating lactoperoxidase system in cheese milk on the quality of Saint-Paulin cheese. Inter. J. Dairy Technol., 61(1):75-83.

[3]Fonteh, F.A., Grandison, A.S., Lewis, M.J., 2005. Factors affecting lactoperoxidase activity. Inter. J. Dairy Technol., 58(4):233-236.

[4]Griffiths, M.W., 1986. Use of milk enzymes as index of heat treatment. J. Food Prot., 49:696-705.

[5]Isobe, N., Kubota, H., Yamasaki, A., et al., 2011. Lactoperoxidase activity in milk is correlated with somatic cell count in dairy cows. J. Dairy Sci., 94(8):3868-3874.

[6]Kumar, R., Bhatia, K.L., 1999. Standardization of method for lactoperoxidase assay in milk. Lait, 79(2):269-274.

[7]Kussendrager, K.D., van Hooijdonk, A.C.M., 2000. Lactoperoxidase: physico-chemical properties, occurrence, mechanism of action and applications. Brit. J. Nutr., 84(1):19-25.

[8]Li, B., Dong, Y., Xu, B., 2012. A spectrophotometric method for determining the activity of lipoxygenase activity in rice bran and its enzymatic characteristics study. J. Chin. Inst. Food Sci. Technol., 12(9):192-197 (in Chinese).

[9]Liu, S.J., Qin, W.Q., Liu, W., et al., 2013. Synergistic mechanism of Tween-20 to oleic acid in bauxite flotation. Chin. J. Nonferrous Met., 23(8):2284-2289 (in Chinese).

[10]Marks, N.E., Grandison, A.S., Lewis, M.J., 2001. Challenge testing of the lactoperoxidase system in pasteurised milk. J. Appl. Microbiol., 91(4):735-741.

[11]Mazri, C., Sánchez, L., Ramos, S.J., et al., 2012. Effect of high-pressure treatment on denaturation of bovine lactoferrin and lactoperoxidase. J. Dairy Sci., 95(2):549-557.

[12]Montiel, R., Bravo, D., de Alba, M., et al., 2012. Combined effect of high pressure treatments and the lactoperoxidase system on the inactivation of Listeria monocytogenes in cold-smoked salmon. Innov. Food Sci. Emerg. Techol., 16:26-32.

[13]Pinho, C.R.G., Franchia, M.A., Tribsta, A.A.L., et al., 2011. Effect of ultra high pressure homogenization on alkaline phosphatase and lactoperoxidase activity in raw skim milk. Procedia Food Sci., 1:874-878.

[14]Pruitt, K.M., Kamau, D.N., Miller, K., et al., 1990. Quantitative, standardized assays for determining the concentrations of bovine lactoperoxidase, human salivary peroxidase, and human myeloperoxidase. Anal. Biochem., 191(2):278-286.

[15]Pütter, J., Becker, R., 1983. Peroxidases. In: Bergmeyer, H.U. (Ed.), Methods of Enzymatic Analysis. Verlag, Florida Chemie, p.286-293.

[16]Seifu, E., Buys, E.M., Donkin, E.F., 2005. Significance of the lactoperoxidase system in the dairy industry and its potential applications. Food Sci. Technol., 16(4):137-154.

[17]Shakeel-ur-Rehman, Farkye, N.Y., 2003. Lactoperoxidase. In: Roginski, R., Fuquay, J.W., Fox, P.F. (Eds.), Encyclopedia of Dairy Science. Academic Press, Amsterdam, p.939-941.

[18]Shimizu, E., Kobayashi, T., Wakabayashi, H., et al., 2011. Effects of orally administered lactoferrin and lactoperoxidase-containing tablets on clinical and bacteriological profiles in chronic periodontitis patients. Inter. J. Dent., 2011:1-9.

[19]Shin, K., Tomita, M., Lönnerdal, B., 2000. Identification of lactoperoxidase in mature human milk. J. Nutr. Biochem., 11(2):94-102.

[20]Shindler, J.S., Childs, R.E., Bradsley, W.G., 1976. Peroxidase from human cervical mucus, the isolation and characterisation. Eur. J. Biochem., 65(2):325-331.

[21]Singh, A.K., Singh, N., Sharma, S., et al., 2008. Crystal structure of lactoperoxidase at 2.4 Å resolution. J. Mol. Biol., 376(4):1060-1075.

[22]Tenovuo, J., 2002. Clinical applications of antimicrobial host proteins lactoperoxidase, lysozyme and lactoferrin in xerostomia: efficacy and safety. Oral Dis., 8(1):23-29.

[23]van Hooijdonk, A.C.M., Kussendrager, K.D., Steijns, J.M., 2000. In vivo antimicrobial and antiviral activity of components in bovine milk and colostrums involved in non-specific defence. Br. J. Nutr., 84(Suppl. 1):S127-S134.

[24]Wolfson, L.M., Sumner, S.S., 1993. Antibacterial activity of the lactoperoxidase system: a review. J. Food Prot., 56(10):887-892.

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