بررسی فراوانی آفلاتوکسین M1 به روش الایزا و کروماتوگرافی مایع با کارایی بالا در شیر خام و پاستوریزه در شهرستان‌های شمال استان کرمان
محورهای موضوعی :
علوم و صنایع غذایی
سمیه صادقی
1
,
حمیدرضا قیصری
2
,
سارا بصیری
3
,
حسین رشیدی
4
,
سید شهرام شکرفروش
5
1 - دانش آموخته دکترای بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران
2 - دانشیار بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران
3 - استادیار بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران
4 - کارشناس ارشد اداره کل دامپزشکی استان کرمان، ایران
5 - استاد بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران
تاریخ دریافت : 1397/08/16
تاریخ پذیرش : 1398/01/31
تاریخ انتشار : 1398/11/01
کلید واژه:
HPLC,
الایزا,
آفلاتوکسین M1,
شیر,
چکیده مقاله :
آفلاتوکسینM1، در شیر دام های تغذیه شده با علوفه آلوده وجود دارد. در مطالعه حاضر 207 نمونه شامل 153 نمونه شیر خام از گاوداری‌های صنعتی دوازده منطقه مختلف استان کرمان و 54 نمونه شیر پاستوریزه تولیدی ده کارخانه فرآوری شیر استان کرمان جمع آوری شد و فراوانی آفلاتوکسین M1 در آن ها با روش الایزا موردسنجش قرار گرفت. نتایج نشان داد که غلظت آفلاتوکسین M1 در 2/41 درصد از شیرهای خام و 2/35 درصد از شیرهای پاستوریزه بیش از محدوده مجاز استاندارد ملی ایران (ng/L100) بود. میزان آفلاتوکسین M1 در شیر پاستوریزه به‌طور معنی داری از شیر خام کمتر بود. به‌منظور تأیید داده های الایزا، 24 نمونه (7/15 درصد) از شیرهای خام و 2 نمونه (7/3 درصد) از شیرهای پاستوریزه با روش کروماتوگرافی مایع با کارایی بالا (HPLC) مورد ارزیابی قرار گرفتند که از این میان، 4 نمونه از نمونه‌های مثبت و 5 نمونه منفی، با HPLC تأیید شدند. مقادیر اندازه گیری شده آفلاتوکسین M1 با دو روش الایزا و HPLC همبستگی معنی‌دار داشت. حساسیت و ویژگی آزمون الایزا به‌ترتیب 100 درصد و 25 درصد تعیین گردید. ازآنجاکه سطح آفلاتوکسین شیر در منطقه بالاتر از محدوده مجاز بود، نیاز به اصلاحات اساسی در مدیریت تغذیه گاوداری های منطقه ضروری به نظر می‌رسد. باوجود این‌که الایزا روشی مناسب در غربالگری آفلاتوکسین است ولی استفاده از روش هایی که افزون بر حساسیت بالا، از ویژگی بهتری نیز برخوردار باشند، در تشخیص فراوانی آفلاتوکسین توصیه می شود.
چکیده انگلیسی:
The aflatoxin M1 (AFM1) is present in the milk of livestock fed on contaminated feed. This study was conducted based on the fact that milk is one of the main sources of aflatoxin contamination in human. In order to record the frequency of AFM1, a total of 207 milk samples, including 153 raw milk samples from 12 different regions of Kerman province and 54 pasteurized milk samples produced in 10 dairy factories in Kerman province was checked by Elisa. Results showed that the concentration of AFM1 in 41.2% of raw and 35.2% of the pasteurized samples was above the maximum acceptable level (100 ng/L) approved by the National Institute of Standard, Iran. The amount of AFM1 in pasteurized milk was significantly lower than that in the raw milk. In order to confirm the results of ELISA, 24 samples of raw milk and 2 samples of pasteurized milk were also evaluated by HPLC, from which, four positive and five negative samples were finally confirmed. Although there was no significant correlation between the number of positive and negative samples in both methods, the concentration of AFM1 was significantly correlated. The sensitivity and the specificity of the ELISA test was respectively 100% and 25%. Since the concentration of aflatoxin was higher than the accepted level, a proper strategy of feeding management of the dairy farms in the province is suggested. In spite of the fact that ELISA is a suitable method for screening of AFM1, using a method with higher specificity is also recommended.
منابع و مأخذ:
· Akrami Mohajeri, F., Amiri, A., Khorramdel Azad, H., Ahmadi, Z., Asadollahi, Z., Rezaeian, M., Fallah, A.A., and Ghalebi. S.R. (2015). Occurrence of Aflatoxin M1 in Raw and Pasteurized Milk Produced in Rafsanjan, Iran. Journal of Community Health Research.;4(3): 215- 219.
· Asi, M.R., Iqbal, S.Z., Ariño, A. and Hussain, A. (2012). Effect of seasonal variations and lactationtimes on aflatoxin M1 contamination in milk of different species from Punjab, Pakistan. Food Control, 25(1): 34-38.
· Barami, A.R., Pour Elmi, M.R., Irani, M. (2011). Contamination levels of aflatoxin M1 in bulk raw milk of Chaloos and Ramsar. Food Hygiene, 4(4): 53- 60. [In Persian]
· Calleri, E., Marrubini, G., Brusotti, G., Massolini, G. and Caccialanza, G. (2007). Development and integration of an immunoaffinity monolithic disk for the onlinesolid-phase extraction and HPLC determination with fluorescence detection of aflatoxin B1 in aqueous solutions. Journal of Pharmaceutical and Biomedical Analysis, 44(2): 396–403.
· Chen, J., Gao, J. (1993). The Chinese total diet study in 1990. Part 1 Chemical contaminants. Journal of AOAC International, 76(6): 1193–1205.
· Choudhary, P.L., Sharma, R.S. and Borkartria, V.N. (1998). Effect of chilling and heating on aflatoxin M1 content of contaminated Indian cow’s milk. Egyptian Journal of DairyScience, 26: 223–229.
· Daraei garmkhany, A., Zighamian, H., Sarhangpour, R., Rasti, M., and Aghajnai, N. (2011). Occurrence of Aflatoxin Ml in Raw and pasteurized milk in Esfahan province of Iran. MINERVA B!OfEC, 23(2-3):53-7.
· Deshpande, S.S. (2002). Fungal toxins. In S. S. Deshpande (Ed.), Handbook of food toxicology New York: Marcel Decker. pp. 387–456.
· Deveci, O. (2007). Changes in the concentration of aflatoxin M1 during manufactureand storage of white pickled cheese. Food Control, 18(9): 1103–1107.
· Gholampour Azizi, I., Khoushnevis, S.H. and Hashemi, S.J. (2007). Aflatoxin M1 level in pasteurized and sterilized milk of Babol city. Tehran University Medical Journal, 65(1): 20-24. [In Persian]
· Hernández-Hierro, J.M., García-Villanova, R.J. and González-Martín, I. (2008). Potential of near infrared spectroscopy for the analysis of mycotoxins applied tonaturally contaminated red paprika found in the Spanish market. Analytica Chimica Acta, 622(1–2): 189–194.
· Institute of Standards and Industrial Research of Iran )ISIRI(, (2016). Milk and milk products – Raw milk – Specification and test methods. Amendment No.1, ISIRI No. 5925. [In Persian]
· Karimi, G., Hassanzadeh, M., Teimuri, M., Nazari, F. and Nili, A. (2007). Aflatoxin M1 Contamination in Pasteurized Milk in Mashhad, Iran. Iranian Journal of Pharmaceutical Sciences, 3(3): 153-156. [In Persian]
· Kim, E.K., Shon, D.H., Ryu, D., Park, J.W., Hwang, H.J. and Kim, Y.B. (2000). Occurrence of aflatoxin M1 in Korean dairy products determined by ELISA and HPLC. Food Additives and Contaminants, 17(1): 59 -64
· Kuiper-Goodman, T. (1999). Approaches to the risk analysis of mycotoxins in thefood supply. Third Joint FAO/WHO/UNEP International Conference on Mycotoxins. Tunis, Tunisia Myc –Conf/99/7a, 4–6 ftp://ftp.fao.org/es/esn/food/myco7a.
· Manetta, A. C., Di Giuseppe, L., Giammarco, M., Fusaro, I., Simonella, A., Gramenzi, A., et al. (2005). High-performance liquid chromatography with post-column derivatisation and fluorescence detection for sensitive determination of aflatoxin M1 in milk and cheese. Journal of Chromatography A, 1083(1–2): 219–222.
· Nile, S.H., Se Won Park, S.W. and Khobragade, C.N. (2015). Occurrence and analysis of aflatoxin M1 in milk produced by Indian dairy species. Food and Agricultural Immunology. 27(3): 355-368 DOI: 10.1080/09540105.2015.1104655.
· Panahi, P., Kasaee, S., Mokhtari, A., Sharifi, A., and Jangjou, A. (2011). Assessment of Aflatoxin M1 Contamination in Raw Milk by ELISA in Urmia, Iran. American-Eurasian Journal of Toxicological Sciences 3 (4): 231-233.
· Pei, S.C., Zhang, Y.Y., Eremin, S.A. and Lee, W.J. (2009). Detection of aflatoxin M1 in milk products from China by ELISA using monoclonal antibodies. Food Control, 20(12): 1080–1085.
· Purchase, I., Steyn, M., Rinsma, R. and Tustin, R. (1978). Reduction of the aflatoxin M1 content of milk of processing. Food and Cosmetics Toxicology, 10(3): 383-387.
· Rastogi, S., Dwivedi, P. D., Khanna, S. K. and Das, M. (2004). Detection of aflatoxin M1 contamination in milk and infant milk products from Indian markets by ELISA. Food Control. 15(4): 287–290.
· Ruangwises, N. and Ruangwises, S. (2010). Aflatoxin M1 contamination in raw milk within the central region of Thailand. Bull Environ Contam Toxicol. 85(2): 195–198.
· Stoloff, L., Van agmond, H.P. and Parks, D.L. (1991). Rationales for the establishment of limits and regulations for mycotoxins. Food Additives and Contaminants, 8(2): 222–231.
· Tajic, H., Razavi Rohani, S.M., and Moradi, M. (2007). Detection of aflatoxin M1 in raw and commercial pasteurized milk in Urmia, Iran. PAKISTAN Journal of Biological Sciences 10(22): 4103- 4107.
· Vagef, R., and Mahmoudi, R. (2013). Occurrence of Aflatoxin M1 in raw and pasteurized milk produced in west region of Iran (during summer and winter). International Food Research Journal 20(3): 1421-1425.
· Var, I., Kabak, B. and Gok, F. (2007). Survey of aflatoxin B1 in helva, a traditional Turkish food, by TLC. Food Control, 18(1): 59–62.
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· Akrami Mohajeri, F., Amiri, A., Khorramdel Azad, H., Ahmadi, Z., Asadollahi, Z., Rezaeian, M., Fallah, A.A., and Ghalebi. S.R. (2015). Occurrence of Aflatoxin M1 in Raw and Pasteurized Milk Produced in Rafsanjan, Iran. Journal of Community Health Research.;4(3): 215- 219.
· Asi, M.R., Iqbal, S.Z., Ariño, A. and Hussain, A. (2012). Effect of seasonal variations and lactationtimes on aflatoxin M1 contamination in milk of different species from Punjab, Pakistan. Food Control, 25(1): 34-38.
· Barami, A.R., Pour Elmi, M.R., Irani, M. (2011). Contamination levels of aflatoxin M1 in bulk raw milk of Chaloos and Ramsar. Food Hygiene, 4(4): 53- 60. [In Persian]
· Calleri, E., Marrubini, G., Brusotti, G., Massolini, G. and Caccialanza, G. (2007). Development and integration of an immunoaffinity monolithic disk for the onlinesolid-phase extraction and HPLC determination with fluorescence detection of aflatoxin B1 in aqueous solutions. Journal of Pharmaceutical and Biomedical Analysis, 44(2): 396–403.
· Chen, J., Gao, J. (1993). The Chinese total diet study in 1990. Part 1 Chemical contaminants. Journal of AOAC International, 76(6): 1193–1205.
· Choudhary, P.L., Sharma, R.S. and Borkartria, V.N. (1998). Effect of chilling and heating on aflatoxin M1 content of contaminated Indian cow’s milk. Egyptian Journal of DairyScience, 26: 223–229.
· Daraei garmkhany, A., Zighamian, H., Sarhangpour, R., Rasti, M., and Aghajnai, N. (2011). Occurrence of Aflatoxin Ml in Raw and pasteurized milk in Esfahan province of Iran. MINERVA B!OfEC, 23(2-3):53-7.
· Deshpande, S.S. (2002). Fungal toxins. In S. S. Deshpande (Ed.), Handbook of food toxicology New York: Marcel Decker. pp. 387–456.
· Deveci, O. (2007). Changes in the concentration of aflatoxin M1 during manufactureand storage of white pickled cheese. Food Control, 18(9): 1103–1107.
· Gholampour Azizi, I., Khoushnevis, S.H. and Hashemi, S.J. (2007). Aflatoxin M1 level in pasteurized and sterilized milk of Babol city. Tehran University Medical Journal, 65(1): 20-24. [In Persian]
· Hernández-Hierro, J.M., García-Villanova, R.J. and González-Martín, I. (2008). Potential of near infrared spectroscopy for the analysis of mycotoxins applied tonaturally contaminated red paprika found in the Spanish market. Analytica Chimica Acta, 622(1–2): 189–194.
· Institute of Standards and Industrial Research of Iran )ISIRI(, (2016). Milk and milk products – Raw milk – Specification and test methods. Amendment No.1, ISIRI No. 5925. [In Persian]
· Karimi, G., Hassanzadeh, M., Teimuri, M., Nazari, F. and Nili, A. (2007). Aflatoxin M1 Contamination in Pasteurized Milk in Mashhad, Iran. Iranian Journal of Pharmaceutical Sciences, 3(3): 153-156. [In Persian]
· Kim, E.K., Shon, D.H., Ryu, D., Park, J.W., Hwang, H.J. and Kim, Y.B. (2000). Occurrence of aflatoxin M1 in Korean dairy products determined by ELISA and HPLC. Food Additives and Contaminants, 17(1): 59 -64
· Kuiper-Goodman, T. (1999). Approaches to the risk analysis of mycotoxins in thefood supply. Third Joint FAO/WHO/UNEP International Conference on Mycotoxins. Tunis, Tunisia Myc –Conf/99/7a, 4–6 ftp://ftp.fao.org/es/esn/food/myco7a.
· Manetta, A. C., Di Giuseppe, L., Giammarco, M., Fusaro, I., Simonella, A., Gramenzi, A., et al. (2005). High-performance liquid chromatography with post-column derivatisation and fluorescence detection for sensitive determination of aflatoxin M1 in milk and cheese. Journal of Chromatography A, 1083(1–2): 219–222.
· Nile, S.H., Se Won Park, S.W. and Khobragade, C.N. (2015). Occurrence and analysis of aflatoxin M1 in milk produced by Indian dairy species. Food and Agricultural Immunology. 27(3): 355-368 DOI: 10.1080/09540105.2015.1104655.
· Panahi, P., Kasaee, S., Mokhtari, A., Sharifi, A., and Jangjou, A. (2011). Assessment of Aflatoxin M1 Contamination in Raw Milk by ELISA in Urmia, Iran. American-Eurasian Journal of Toxicological Sciences 3 (4): 231-233.
· Pei, S.C., Zhang, Y.Y., Eremin, S.A. and Lee, W.J. (2009). Detection of aflatoxin M1 in milk products from China by ELISA using monoclonal antibodies. Food Control, 20(12): 1080–1085.
· Purchase, I., Steyn, M., Rinsma, R. and Tustin, R. (1978). Reduction of the aflatoxin M1 content of milk of processing. Food and Cosmetics Toxicology, 10(3): 383-387.
· Rastogi, S., Dwivedi, P. D., Khanna, S. K. and Das, M. (2004). Detection of aflatoxin M1 contamination in milk and infant milk products from Indian markets by ELISA. Food Control. 15(4): 287–290.
· Ruangwises, N. and Ruangwises, S. (2010). Aflatoxin M1 contamination in raw milk within the central region of Thailand. Bull Environ Contam Toxicol. 85(2): 195–198.
· Stoloff, L., Van agmond, H.P. and Parks, D.L. (1991). Rationales for the establishment of limits and regulations for mycotoxins. Food Additives and Contaminants, 8(2): 222–231.
· Tajic, H., Razavi Rohani, S.M., and Moradi, M. (2007). Detection of aflatoxin M1 in raw and commercial pasteurized milk in Urmia, Iran. PAKISTAN Journal of Biological Sciences 10(22): 4103- 4107.
· Vagef, R., and Mahmoudi, R. (2013). Occurrence of Aflatoxin M1 in raw and pasteurized milk produced in west region of Iran (during summer and winter). International Food Research Journal 20(3): 1421-1425.
· Var, I., Kabak, B. and Gok, F. (2007). Survey of aflatoxin B1 in helva, a traditional Turkish food, by TLC. Food Control, 18(1): 59–62.
