Authenticating Emergent Adulterant 6-Benzylaminopurine in Bean Sprouts: Virtual Hapten Similarity Enhanced Immunoassay.

6-Benzylaminopurine (6-BA), a plant growth regulator with cytokinin-like properties, was recently reported to be illegally used in bean sprouts to increase their commercial appearance. It is still nevertheless challenging to quickly detect this adulteration. In this work, four novel haptens (haptens 1-4) of 6-BA were rationally designed with computer-assisted modeling analysis and then synthesized for use as immunizing haptens to produce antibodies. One of two obtained antibodies showed high sensitivity and specificity toward 6-BA. Based on the most sensitive anti-6-BA antibody, an indirect competitive enzyme-linked immunosorbent assay (icELISA) was performed, which demonstrated a 50% inhibition concentration (IC50) of 1.18 µg/L and a limit of detection of 0.075 µg/L. The average recoveries of this icELISA for 6-BA of spiked samples ranged from 87.2 to 95.0% with a coefficient of variation of less than 8.7%. Furthermore, the blind samples were detected simultaneously by the method and HPLC-MS/MS, and the results showed good agreement with each other. Therefore, the proposed icELISA can facilitate the rapid surveillance screening of adulterated 6-BA in sprout vegetables.

Identifying fungicide difenoconazole as illegal growth regulator in vegetable: Computer-aided hapten similarity to enhance immunoassay sensitivity.

Difenoconazole, a fungicide with broad-spectrum properties, has recently been found to have been used illegally used as a plant growth regulator in Brassica campestris, with the intent of inducing thick stems and dark green leaves. However, analysts have encountered challenges in implementing a rapid surveillance screening approach for this purpose. In this study, a novel hapten was designed to improve the analytical performance of difenoconazole immunoassay. Specifically, the triazole of the original hapten was replaced with a benzene ring, guided by molecular simulation. This led to the development of a very sensitive antibody and the subsequent development of a competitive indirect enzyme linked immunosorbent assay (ciELISA) for the detection of difenoconazole in vegetable samples. The assay exhibited a working range of 0.16 ng mL-1 to 9.64 ng mL-1, with a detection limit of 0.05 ng mL-1. Upon analysis of blind samples, a strong correlation was observed between the ciELISA and HPLC-MS/MS methods. As a result, the proposed technique may prove to be an excellent tool for the rapid detection of difenoconazole overuse and adulteration in vegetables.

Developing an ultrasensitive immunochromatographic assay for authentication of an emergent fraud aminopyrine in herbal tea.

Aminopyrine is a nonsteroidal anti-inflammatory drug only for medical purposes, however, it has been illegally added in traditional Chinese herbal teas for fraud activity recently. In this study, a specific antibody against aminopyrine with IC50 of 3.00 ng/mL was obtained for the first time by a rational hapten design. Furthermore, an ultrasensitive gold nanoparticles immunochromatographic assay (AuNPs-ICA) for determination of aminopyrine based on a portable reader was firstly developed, with cut-off value of 100.00 ng/mL, limit of detection (LOD) of 4.80 ng/mL and limit of quantification (LOQ) of 5.71 ng/mL for herbal tea, respectively. The recovery rates ranged from 93.21 % to 105.61 %, with inter-assay coefficient of variation (CV) from 1.08 % to 3.82 %. Additionally, 24 blind samples were examined simultaneously by AuNPs-ICA and LC-MS/MS, demonstrating a good consistency for each other. The proposed AuNPs-ICA is an ultrasensitive and reliable tool for on-site surveillance screening of fraud additives in herbal tea.