A comparison of TLC-densitometry and HPLC method for the determination of biogenic amines in fish and fishery products

Defence Food Research Laboratory, Mysore 570011, India
Food Chemistry (Impact Factor: 3.39). 11/2001; 75(2):255-259. DOI: 10.1016/S0308-8146(01)00173-X


A comparative study on the suitability of chromatographic techniques such as thin layer chromatography (TLC)-densitometry and high performance liquid chromatography (HPLC) for the analysis of biogenic amines in fish and fishery products was carried out. The HPLC method was found to offer a good linearity (5–100 ng), sensitivity and repeatability (<3%), but required sophisticated instrumentation, technical skill and high operational cost and time. On the other hand, with a little loss in linearity (20–300 ng) and repeatability (<8%), TLC-densitometry was found to be rapid and less expensive. In addition, this method is suitable for rapid and simultaneous screening of several samples at a time. Therefore, the TLC-densitometric method can be effectively used in the fish industry to detect biogenic amines, especially the toxic histamine, and putrescine and cadaverine, which can potentiate histamine toxicity in fish and fishery products.

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    • "This technique has been successfully employed for analysis of flavonoids and phenolic acids in wine, propolis, and different medicinal plants [4]. Several studies comparing TLC and HPLC have shown that there is no statistical difference between these methods, and advantage due to lower cost is given to TLC [5] [6]. Determination of xanthines as the main constituents is time consuming titration [7]. "
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    ABSTRACT: Raw material, different formulations of foods, and dietary supplements of mate demands control of the content of bioactive substances for which high performance thin layer chromatography (TLC), described here, presents simple and rapid approach for detections as well as quantification. Using TLC densitometry, the following bioactive compounds were identified and quantified: chlorogenic acid (2.1 mg/g), caffeic acid (1.5 mg/g), rutin (5.2 mg/g), quercetin (2.2 mg/g), and kaempferol (4.5 mg/g). The results obtained with TLC densitometry for caffeine (5.4 mg/g) and theobromine (2.7 mg/g) show no statistical difference to the content of total xanthines (7.6 mg/g) obtained by UV-Vis spectrophotometry. Thus, TLC remains a technique of choice for simple and rapid analysis of great number of samples as well as a primary screening technique in plant analysis.
    Full-text · Article · Jun 2013 · Journal of Analytical Methods in Chemistry
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    • "In contrast to many of the other more potent seafood toxins, the relatively high action levels established for histamine in fish allow for its detection by a variety of different approaches ranging from simple and inexpensive thin layer chromatography (TLC) procedures to resource-intensive and more powerful liquid chromatographymass spectrometry (LC-MS) methods (Hungerford, 2010). Most of the histamine separation methods applied in fish use reversedphase high performance liquid chromatography (HPLC) with detection schemes based on pre-column derivatization (Mietz and Karmas, 1978; Hui and Taylor, 1983; Malle et al., 1996) or post-column derivatization (Glória et al., 1999; Brillantes and Samosorn, 2001) to produce fluorescent products or strong chromophores , but direct UV detection of histamine imidazole ring has also been applied (Shakila et al., 2001; Cinquina et al., 2004b). "
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    ABSTRACT: The presence of biogenic amines (BAs) in raw and processed seafood, associated with either time/temperature conditions or food technologies is discussed in the present paper from a safety and prevention point of view. In particular, storage temperature, handling practices, presence of microbial populations with decarboxylase activity and availability of free amino acids are considered the most important factors affecting the production of BAs in raw seafood. On the other hand, some food technological treatments such as salting, ripening, fermentation, or marination can increase the levels of BAs in processed seafood. The consumption of high amount of BAs, above all histamine, can result in food borne poisoning which is a worldwide problem. The European Regulation established as maximum limits for histamine, in fishery products from fish species associated with high histidine amounts, values ranging from 100 to 200 mg/kg, while for products which have undergone enzyme maturation treatment in brine, the aforementioned limits rise to 200 and 400 mg/kg. Preventive measures and emerging methods aiming at controlling the production of BAs are also reported for potential application in seafood industries.
    Full-text · Article · Jun 2012 · Frontiers in Microbiology
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    • "Frattini and Lionetti, 1998; Shakila et al., 2001, Cinquina et al.,2004b). Other popular separation-based methods include ion chromatography (Cinquina et al.,2004a), capillary electrophoresis (Gallardo et al., 1997; Zhang and Sun, 2004), paper electrophoresis (Sato et al., 2002, 2006), thin layer chromatography (Lieber and Taylor, 1978; Bajc and Gačnik, 2009) and gas chromatography-mass spectrometry (Marks and Anderson, 2006). "
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    ABSTRACT: Scombroid poisoning, also called histamine fish poisoning, is an allergy-like form of food poisoning that continues to be a major problem in seafood safety. The exact role of histamine in scombroid poisoning is not straightforward. Deviations from the expected dose-response have led to the advancement of various possible mechanisms of toxicity, none of them proven. Histamine action levels are used in regulation until more is known about the mechanism of scombroid poisoning. Scombroid poisoning and histamine are correlated but complicated. Victims of scombroid poisoning respond well to antihistamines, and chemical analyses of fish implicated in scombroid poisoning generally reveal elevated levels of histamine. Scombroid poisoning is unique among the seafood toxins since it results from product mishandling rather than contamination from other trophic levels. Inadequate cooling following harvest promotes bacterial histamine production, and can result in outbreaks of scombroid poisoning. Fish with high levels of free histidine, the enzyme substrate converted to histamine by bacterial histidine decarboxylase, are those most often implicated in scombroid poisoning. Laboratory methods and screening methods for detecting histamine are available in abundance, but need to be compared and validated to harmonize testing. Successful field testing, including dockside or on-board testing needed to augment HACCP efforts will have to integrate rapid and simplified detection methods with simplified and rapid sampling and extraction. Otherwise, time-consuming sample preparation reduces the impact of gains in detection speed on the overall analysis time.
    Full-text · Article · Feb 2010 · Toxicon
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