Development of an Enzyme Immunoassay Using a Monoclonal Antibody against the Psychoactive Diterpenoid Salvinorin A

Abstract

Salvinorin A (1), the main active constituent in Salvia divinorum, is a highly selective kappa-opioid receptor agonist with hallucinogenic effects, which is regulated in several countries. In the present study, a monoclonal antibody (mAb) against 1 was prepared, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) system was developed for the detection of salvinorins. To raise mAbs against 1, salvinorin B (2) hemisuccinate was synthesized and used to prepare the immunogen 2-bovine serum albumin conjugate. This technique was used to prepare a hybridoma cell line, 3D5, which secreted a mAb that recognized 1. The mAb was shown to have specificity for 1 and other salvinorins in cross-reactivity tests. The intra-assay calibration range by icELISA using the mAb against 1 was 0.0195-0.625 μg/mL. After validating the icELISA using intra- and interassays, a recovery experiment and analysis of several plants in the family Lamiaceae, including S. divinorum, confirmed that the analytical method based on ELISA is not only simple but also precise, accurate, sensitive, and sufficiently reliable. The results indicate that icELISA is a useful tool in the identification of S. divinorum.

Quantitative Fluorescence Quenching on Antibody-conjugated Graphene Oxide as a Platform for Protein Sensing

“…However, the accuracy of ELISA is affected by a few shortcomings intrinsic to the technique. For example, the uncertainty of the recognition between antibody and antigens could cause false signals123 the ELISA plates made of polystyrene lack definitive chemical surface properties to provide chemical binding of the protein4; and intricate procedures such as “plate cleaning” increase opportunities for systemic and human errors. In recent years, many research groups have tried to improve the accuracy of ELISA by synthesizing new ELISA polymer substitutes4, introducing nanomaterials56, employing more sensitive biomarkers7, and applying monoclonal antibodies8.…”

An overview of recent developments in the analytical detection of new psychoactive substances (NPSs)

“…137 The use of this plant as a psychoactive substance has spread globally, its major constituentsalvinorin A (SA) is a known selective opioid antagonist and to this end emphasis in the literature has been put on detecting SA. 138 Recently studies have postured SAs effects involve the endocannabinoid system. 138 Recently studies have postured SAs effects involve the endocannabinoid system.…”

Salvinorina A: terpeno alucinógeno presente en Salvia divinorum Epling & Játiva

“…Esta especie pertenece a la familia Lamiaceae, también conocida como "familia de la menta", caracterizada por tener variedad de productos metabólicos secundarios que a su vez se usan en varias partes del mundo, por ejemplo, en condimentos y en aceites esenciales y ornamentales [9][10][11][12][13][14]. Por su parte, los indígenas Mazatecos usan la planta en "prácticas espirituales, de adivinación y de curación", las cuales consisten en predicción de lluvias, corroboración de buenas cosechas, y diagnóstico y curación de enfermedades como la anemia, la diarrea y los dolores de cabeza [4,6,[15][16][17]. Todas las poblaciones nativas registradas de Salvia divinorum E&J son originarias de Oaxaca, en el sur de México.…”

“…En Noruega, Finlandia, Estonia e Islandia, es legal el uso de la planta con fines medicinales y solo puede ser obtenida con prescripción médica. Por ejemplo, se utiliza para el tratamiento de personas dependientes a la cocaína y a la heroína [4,8,9,15,[20][21][22][23][24][25][26].…”