مقایسه روش الکتروفورز موئینه ای با آشکارساز جذبی با روش کروماتوگرافی مایع با عملکرد بالا برای اندازه گیری مقدار هیستامین در محیط کشت میکروبی

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری تخصصی بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران

2 استاد بخش بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران

3 دانشیار بخش بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران

چکیده

هیستامین یکی از شاخص­های مهم فساد باکتریایی در مواد غذایی است که می­تواند موجب مسمومیت در مصرف‌­کنندگان شود. بنابراین شناسایی و اندازه­گیری مقدار هیستامین در مواد غذایی از اهمیت به‌سزایی برخوردار است. در این تحقیق، دو روش کروماتوگرافی مایع با عملکرد بالا (HPLC) به‌عنوان روش مرجع و الکتروفورز موئینه­ای با آشکار­ساز جذبی (CZE) برای اندازه­گیری مقدار هیستامین تولیدی در محیط کشت TSB با هم مقایسه شدند. برای این منظور محیط کشت با سوش استاندارد مولد هیستامین استافیلوکوکوس اپیدرمیدیس TYH1 و دو سوش استافیلوکوکوس کاپیتیس و استافیلوکوکوس کارنوسوس دارای ژن هیستیدین دکربوکسیلاز تلقیح شد. زمان مهاجرت در روش CZE و زمان بازداری در روش HPLC برای هیستامین به‌ترتیب 4/5 و 0/12 دقیقه به‌دست آمد. منحنی استاندارد در محدوده µg/ml 200- 25/6 به‌صورت خطی با هر دو با معادله رگرسیون y=0.000004x (r2=0.999) به‎دست آمد. مقدار هیستامین تولید شده توسط سه سوش باکتری و اندازه­گیری شده با دو روش مذکور تفاوت آماری معنی­داری نداشت و ضریب همبستگی بین یافته­های به‌دست ‌آمده از دو روش 99/0 بود. با توجه به یافته­های مشابه دو روش، روش CZE به‌دلیل عدم نیاز به آماده‌سازی نمونه، سادگی، حساسیت و کم‌هزینه بودن، به‌عنوان روشی مناسب برای اندازه­گیری مقدار هیستامین در محیط­های کشت میکروبی پیشنهاد می­گردد.

کلیدواژه‌ها


عنوان مقاله [English]

Comparison of capillary zone electrophoresis and high performance liquid chromatography for the determination of histamine in bacterial culture media

نویسندگان [English]

  • S. Pashangeh 1
  • S.S. Shekarforoush 2
  • M. Aminlari 2
  • S. Hosseinzadeh 3
1 . Ph.D. Graduate of Food Hygiene, Department of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
2 Department of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
3 Department of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
چکیده [English]

The presence of histamine in foods is an important indicator of food spoilage and can also cause food poisoning in consumers. Therefore, monitoring of histamine level in food is important. This paper has focused on the comparing of capillary zone electrophoresis (CZE) with HPLC for the determination of histamine in tryptic soy broth. For this reason, the culture media was inoculated with a standard strain of histamine-producing Staphylococcus epidermidis TYH1, as well as S. capitis and S. carnosus strains harboring histidine decarboxylase-hdc. The migration time of histamine in CZE assay and retention time of HPLC were found 5.4 and 12 min, respectively. The calibration graph was linear ranged 6.25 to 200 μg/ml which was estimated using the regression equation of y=0.000004x (r2=0.999) for both methods. Determination of histamine produced by these three strains indicated no significant differences using CZE with HPLC methods. The results show that the CZE is suitable for the determination of histamine in bacterial culture, since, there is no need for initial preparation, simplicity, sensitivity and low cost.

کلیدواژه‌ها [English]

  • Histamine
  • Capillary electrophoresis
  • High-performance liquid chromatography
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