Immunochromatographic assay for the detection of kwakhurin and its application for the identification of Pueraria candollei var. mirifica (Airy Shaw & Suvat.) Niyomdham

Abstract

Introduction: The plant Pueraria candollei var. mirifica (Airy Shaw & Suvat.) Niyomdham (PM), known by its common Thai name as white Kwao Krua, is sometimes misidentified because it presents similar botanical characteristics to those of Butea superba (red Kwao Krua). The phytochemicals in PM are phytoestrogens in the class of isoflavonoids, but Butea superba contains flavonoids that exhibit androgenic and antiestrogen effects.Objectives: This research aims to develop a simple analytical method for identification and to differentiate PM from red Kwao Krua and other Pueraria species.Methods: A gold nanoparticle-based immunochromatographic assay (ICA) was developed for the detection of kwakhurin (Kwa), a unique compound found in PM. The parameters, including sensitivity, accuracy, precision, and specificity, were validated. All samples were analyzed using ICA and high-performance liquid chromatography with UV detector (HPLC-UV). The results of the two methods were compared for consistency checking. Results:The cutoff limit of Kwa detection was 160 ng/mL, which was lower than in the HPLC-UV method. The repeatability and reproducibility of the ICA preparation and assembly showed high precision. The cross-reactivity to related isoflavonoids was less than 0.32%, which implied high specificity of the ICA for Kwa. Moreover, false-positive and false-negative results from other plant extracts were not observed. Conclusion:The developed ICA is applicable for distinguishing PM from red Kwao Krua and other Pueraria species. This simple analytical method can be applied for the identification of raw PM materials in the industrial and agricultural sectors.

Development and characterisation of highly specific monoclonal antibody‐based immunoassays for the detection and quantification of genistein‐7‐O‐[α‐rhamnopyranosyl‐(1→6)]‐β‐glucopyranoside in Derris scandens (Roxb.) Benth.

“…The LOD of LFIA using mAbs for kwakhurin and miroestrol was reported to be 160 ng/mL and 1000 ng/mL, respectively. 15,26 The LOD observed for this strip was similar to that in earlier investigations.…”

“…This LFIA method could detect sennoside A, 13 sennoside B, 13 miroestrol, 14 and kwakhurin. 15 Additionally, the use of LFIA for a semi-quantitative study of artesunate has been reported. 16 This study aimed to generate a hybridoma clone that could secrete a GTG-specific mAb.…”

Lateral flow immunoassay for small-molecules detection in phytoproducts: a review

“…The compounds with carboxylic acid and hydroxyl functional groups are usually conjugated by a carbodiimidemediated reaction [33,34,47,[95][96][97][98][99][100][101][102], whereas compounds with active hydrogen, e.g., α-picolines, ketones, esters, and acetylenes, can be conjugated to an amine or amide group of the carrier by a Mannich reaction [103,104]. Furthermore, structure modifications of the parent compounds to add the reactive functional group to the molecule can be conducted when necessary.…”

“… [ 47 ] Aconitine Alkaloid Aconitum spp. [ 29 ] Rhein Anthraquinone Rheum officinale [ 42 ] Kwakhurin Isoflavonoids Pueraria mirifica [ 102 ] Higenamine Benzyltetrahydroisoquinoline alkaloid Aconitum carmichaelii, Asarum heterotrophies, Nandina domestica [ 34 ] Amarogentin Secoiiridoid glycoside Swertia spp., Gentiana spp. [ 46 ] Colchicine Alkaloid Colchicum autumnale [ 44 ] …”

Bioimprinting as a Receptor for Detection of Kwakhurin

“…In our previous study, we focused on Kwa as a marker compound for standardization as it is a phytoestrogen [ 13 ] produced solely by P. candollei . We also developed various immunoassays for the determination of Kwa in P. candollei -derived products [ 15 , 16 , 17 , 18 ] as immunoassays enable sensitive and selective analyses of small molecules [ 19 , 20 ]. However, an animal-based approach is necessary for the production of antibodies used in immunoassays.…”