Quantitative Analysis of Aflatoxins by High-Performance Thin-Layer Chromatography Utilizing a Scientifically Operated Charge-Coupled Device Detector

Abstract

The use of a scientifically operated charge-coupled device (CCD) for the detection and quantitation of aflatoxins on a high-performance thin-layer chromatographic (HPTLC) plate was investigated. A nebulizer-based sample application system was utilized to quantitatively transfer the sample onto the HPTLC plate without disturbing the sorbent layer. Fluorescence excitation of the aflatoxins was accomplished with an ultraviolet transilluminator, which caused the analytes of interest to emit in the bluegreen portion of the visible spectrum. The dynamic range, sensitivity, accuracy, and precision of the system were evaluated. Detection limits of the aflatoxins were determined to be in the low picogram range. The existence of aflatoxins in peanut butter samples was analyzed both quantitatively and qualitatively by using multidimensional TLC. The capabilities of the system for performing realtime monitoring of the separation process were also investigated.Aflatoxins are toxic, extremely carcinogenic substances excreted by green powdery molds known as Aspergillus flavus and Aspergillus parisiticus (Figure 1). These compounds have been found on a variety of agricultural products, livestock feeds, and commercial foodstuffs, including corn, peanuts, cottonseeds, grains, legumes, and tree nuts, and indirectly in dairy products, poultry, and meat products. Aflatoxins were first discovered to be of serious concern in the 1960s, when ducklings, calves, and some 100 000 young turkeys died after ingesting contaminated peanut meal. 1,2 Aflatoxins have been shown to cause cancer of the liver, colon, and kidney in some animals, such as rats, ducks,

Development of a smartphone-based biomimetic sensor for aflatoxin B1 detection using molecularly imprinted polymer membranes

“…A number of traditional instrumental analytical [5,6] as well as immunochemical [6][7][8][9] methods were developed for aflatoxin detection. Chromatographic methods, such as TLC [10], HPLC [11] and LC-MS [7] as well as ELISA-based commerciallyavailable test-systems, are the most widely used analytical methods for detection of aflatoxin B1.…”

Qualitative and Quantitative Analysis of Mycotoxins

“…Modern HP‐TLC is a precise and accurate analytical tool with efficiency, which is comparable to that of HPLC and ELISA methods. Therefore, quantitative and qualitative analysis of aflatoxins has been developed using multidimensional–HPTLC–fluorescence excitation, such as applied in peanut butter samples by Liang and others (1996). Recently, Toteja and others (2006) determined aflatoxin B1 of rice samples, using HPTLC after extraction with water/chloroform and silica gel column cleanup.…”

“…TLC was an extremely powerful, rapid, and low‐cost separation technique in mycotoxicology before HPLC techniques became popular. Several TLC methods were developed for mycotoxin quantitation (Le‐Tutor and Tantaoui Elaraki 1984; Liang and others 1996; Aziz and others 1998; Odhav and Naicker 2002; Rizzo and others 2004; Caldas and Silva 2007) and qualitative determination (Grabarkiewicz‐Szczesna and others 1985; Liang and others 1996; Abrunhosa and others 2001; Odhav and Naicker 2002). Alhough there are some reports on comparable results between TLC and HPLC for the dtermination of aflatoxin M1 in raw, pasteurized, and UHT milk (recovery values ranged from 85.83% to 73.86% at levels of 0.010 to 0.50 μg/L) (Shundo and Sabino 2006) and aflatoxins (B1, B2, G1, and G2) in products with quantification limit (LOQ) of 2 μg/kg, these TLC methods are generally suitable for qualitative analysis at best (Caldas and Silva 2007).…”

Photometric procedure for quantitative analysis of Aflatoxin B1 in peanuts by thin-layer chromatography using charge coupled device detector

“…17,18 By contrast, TLC is a simple and robust technique, 19 which is relatively inexpensive to establish in a testing laboratory, and most laboratories in developing countries have considerable expertise and experience with it. 20 The major problem in TLC method is the quantification that usually is done by visual comparison, however other techniques can be employed to improve the analytical performance, such as densitometry. 21 Charge-coupled devices (CCDs) are two-dimensional detectors containing an array of sensors that can image an area in fraction of seconds or real time.…”

“…These features have made the CCD an excellent detector for many imaging applications in chemical analysis, such as fluorescence detection. 19,20 ImageJ is one of the many image processing tools available and is required for the analysis of images generated by the CCD camera and to carry out the necessary operations. It is a versatile software, which offers a wide array of functionalities that can be customized to meet specific requirements.…”