Determination of hexoestrol residues in animal tissues based on enzyme-linked immunosorbent assay and comparison with liquid chromatography–tandem mass spectrometry

Xu, Chuanlai; Peng, Chifang; Liu, Liqiang; Wang, Liying; Jin, Zhengyu; Chu, Xiaogang

doi:10.1016/j.jpba.2006.01.016

Cited by 17 publications

(5 citation statements)

References 19 publications

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“…DES was able to compete with the coating antigen to interact with the antibody in the range of 50-1350 pg mL -1 , and in this range the absorbance value exhibited a good linear relationship with a logarithmic DES concentration (R 2 = 0.993; Figure 5). Based on the preparation of the polyclonal antibody against DES, Xu et al [16,17] established a direct ELISA method for DES detection. A DES-HRP conjugate was required to perform the direct ELISA assay.…”

Section: Antibody Evaluationmentioning

confidence: 99%

“…Wang et al [14,15] produced two kinds of similar polyclonal antibodies against DES with IC 50 of 1.02 and 1.89 μg L -1 , respectively. Xu et al [16,17] also prepared a polyclonal antibody against DES with an average IC 50 value of 2.4 ng mL -1 , a calibration range of 0.2-30.5 ng mL -1 , and a detection limit of 0.07 ng mL -1 . Compared with the polyclonal antibody, the application of a monoclonal antibody (MAb) was advantageous in terms of purity, sensitivity and specificity.…”

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confidence: 99%

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Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Meng

He³

et al. 2011

Chin. Sci. Bull.

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Based on the preparation of an anti-diethylstilbestrol (DES) monoclonal antibody, a simple and convenient indirect competitive enzyme-linked immunosorbent assay (ELISA) method for DES detection has been developed. The monoclonal antibody demonstrated high sensitivity to DES with an IC 50 value of 275 pg mL -1 and detection limit (LOD) of 90 pg mL -1 . The specificity of the assay was studied by measuring cross-reactivity of the antibody with structurally related compounds of ethinyl estradiol (<7%), estrone (<0.1%), estriol (<0.1%), and diethylstilbestrol benzoate (<0.1%). Chicken, fish, shrimp, urine and bile spiked with different concentration of DES were detected by the developed method, and the recovery rates were greater than 79.5%. Intra-and inter-assay variations were about 6%. This method exhibited high stability with a coefficient of variation less than 10% in buffer and in real samples. The LODs in fish/shrimp, liver, feed and urine spiked with DES were 600, 600, 4800 and 600 pg mL -1 , respectively. These results confirmed that the antibody to DES was successfully produced and could be used to establish ELISA methods for DES detection in food producing animals. Diethylstilbestrol (DES; Figure 1) is an orally active synthetic non-steroid estrogen. Since the 1960s, DES has been used as a growth-promoting agent in livestock [1,2]. Later, DES was found to cause cancer and its use was "phased out in the 1970s" by the FAO/WHO. In 1971, DES was reported to be a teratogen when given to pregnant women and has been associated with a rare vaginal cancer in female offspring [3]. On February 5, 1975, the US Food and Drug Administration (FDA) ordered 25 and 100 mg tablets of DES withdrawn; however, it was still illegally used in the livestock industry.*Corresponding authors (email: xirimo2000@yahoo.com; jintliu@sdu.edu.cn)More than 30 years of researches have confirmed that DES is a teratogen, which can cause malformations of an embryo or fetus. Prenatal exposure to DES is associated with an increased incidence of fibroids of adulthood [4,5]. Therefore, the Ministry of Agriculture of the People's Republic of China issued a banned list (Announcement No. 235; http://www. agri.gov.cn/ zcfg/bmgz/ t20060123_540865.htm, accessed on October 24, 2002) of veterinary drugs and announced that the use of DES in food and animal feed was prohibited. The European Union (EU) requires that residual levels of DES in edible animal food should not exceed 2 ng mL -1 [6]. Consequently, a rapid screening method with high sensitivity and

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Section: Antibody Evaluationmentioning

confidence: 99%

mentioning

confidence: 99%

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Meng

He³

et al. 2011

Chin. Sci. Bull.

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“…However, these methods require extensive sample preparation, expensive instruments and professional operation. Alternatively, enzyme-linked immunosorbent assay (ELISA) has been successfully developed for screening HES in urine or tissues [15,16], but, ELISA also needs incubation and washing steps and is mainly confined to laboratories. Since the development of the lateral flow immunochromatographic strip test (LFIST) for on-site detection of the concentrations of biotech crops in samples of ground grain [17], it has become a more popular method for the qualitative or semi-quantitative detection of various analytes [18][19][20][21][22] for its advantages of a one-step process (only addition of specimen), rapidity (less than 5 min) and costeffectiveness [23,24].…”

Section: Introductionmentioning

confidence: 99%

A lateral flow immunochromatographic strip test for rapid detection of hexoestrol in fish samples

et al. 2018

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A lateral flow immunochromatographic strip test was developed for rapid and sensitive on-site detection of hexoestrol (HES) residues in fish samples with colloidal gold labelling of the anti-HES monoclonal antibody. The strip is composed of a sample pad, a conjugate reagent pad, an absorbent pad and a test membrane containing a control line and a test line. The sensitivity (half inhibitory concentration, IC50) of the strip in the detection of fish extract samples was confirmed to be 1.86 µg kg−1, and the limit of detection value was 0.62 µg kg−1. For intra-assay and inter-assay reproducibility, recoveries of HES-spiked samples ranged from 86.3% to 92.3% and 85.8% to 93.4%, coefficients of variation were 2.91–4.64% and 4.24–5.17%, respectively. High-performance liquid chromatography was employed to confirm the performance of the strip. The strip test takes less than 10 min, and thus provides a repaid method for on-site detection of HES residues.

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“…The common techniques used to detect defective foods are liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry. Nevertheless, these methods are not suitable for on-site analysis (Wang et al, 2016;Xu et al, 2006); moreover, they are expensive and require more skilled personnel to operate them. Therefore, rapid, sensitive, inexpensive and accurate simple and time-saving methods such as Lateral immunochromatographic strips and indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), which require less knowledge for operation, must be developed complementary to conventional instrumental techniques Tochi et al, 2015;Xu et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Development of an immunochromatographic assay for hexestrol and diethylstilbestrol residues in milk

Mukunzi

Tochi

Isanga

et al. 2016

Food and Agricultural Immunology

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Hexestrol (HES) and Diethylstilbestrol (DES) are synthetic hormones, which have been found abuse use in animal-origin food production. We developed an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and immune-chromatographic strip to detect these compounds based on monoclonal antibody. Under optimized conditions, the half-inhibition concentration (IC 50 ) values of ic-ELISA were 0.15 µgL −1 and 0.23 µgL −1 for HES and DES, respectively. Spiked milk samples were analyzed. The recovery of both synthetic hormones in the milk samples was 60.48-102.19% (HES) and 89.34-100.16% (DES). In immunechromatographic assay, the visual cutoff values at 0.5 and 1.0 µgL −1 respectively for HES and DES, which allow us to detect milk samples of a concentration low to 10 µgL −1 for HES and 15 µgL −1 for DES. ARTICLE HISTORY

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Determination of hexoestrol residues in animal tissues based on enzyme-linked immunosorbent assay and comparison with liquid chromatography–tandem mass spectrometry

Cited by 17 publications

References 19 publications

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

A lateral flow immunochromatographic strip test for rapid detection of hexoestrol in fish samples

Development of an immunochromatographic assay for hexestrol and diethylstilbestrol residues in milk

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