Four Novel Phenanthrene Derivatives with <i>α</i>-Glucosidase Inhibitory Activity from <i>Gastrochilus bellinus</i>

Four new phenanthrene derivatives, gastrobellinols A-D (1–4), were isolated from the methanolic extract of Gastrochilus bellinus (Rchb.f.) Kuntze, along with eleven known phenolic compounds including agrostophyllin (5), agrostophyllidin (6), coniferyl aldehyde (7), 4-hydroxybenzaldehyde (8), agrostophyllone (9), gigantol (10), 4-(methoxylmethyl)phenol (11), syringaldehyde (12), 1-(4′-hydroxybenzyl)-imbricartin (13), 6-methoxycoelonin (14), and imbricatin (15). Their structures were determined by spectroscopic methods. Each isolate was evaluated for α-glucosidase inhibitory activity. Compounds 1, 2, 3, 7, 9, 13, and 15 showed higher activity than the drug acarbose. Gastrobellinol C (3) exhibited the strongest α-glucosidase inhibition with an IC50 value of 45.92 μM. A kinetic study of 3 showed competitive inhibition on the α-glucosidase enzyme. This is the first report on the phytochemical constituents and α-glucosidase inhibitory activity of G. bellinus

Seasonal and Geographic Variation in Alkaloid Content of Kratom (<i>Mitragyna speciosa</i> (Korth.) Havil.) from Thailand

The objective of this study was to obtain data on the distribution of alkaloids in kratom plants grown in Thailand. Two collections were performed, covering the southern, central, and northern regions of Thailand and different seasons. The contents of alkaloids, including mitragynine (MG), paynantheine (PAY), and speciogynine (SG), were determined using the validated HPLC method. The 134 samples in the first collection were collected from Nam Phu subdistrict, Ban Na San, Surat Thani, Thailand, during June and October 2019 and January 2020. The maximum mitragynine content was 4.94% w/w in June (late summer), and the minimum content was 0.74% w/w in October (rainy season). To expand the study area after kratom decriminalization, 611 samples were collected in June–August 2021, October–December 2021, and January–April 2022. The accumulation of MG ranged from 0.35 to 3.46% w/w, 0.31 to 2.54% w/w, and 0.48 to 2.81% w/w, respectively. The meteorological data supported the climate’s effect on alkaloid production. Soil analysis revealed the importance of Ca and Mg in promoting alkaloid production. Geographical locations played a role in the variation of MG in kratom leaves, but did not affect the color of leaf veins. In conclusion, the present study suggested that the alkaloid content in kratom diverges based on seasonal and geographical origin

Analysis of canthin-6-one alkaloids derived from Eurycoma spp. by micellar liquid chromatography and conventional high-performance liquid chromatography: a comparative evaluation

Extracts of Eurycoma longifolia Jack (EL) and Eurycoma harmandiana Pierre (EH) contain numerous bioactive compounds and varying matrices that are challenging to separate using chromatographic techniques. Herein, micellar liquid chromatography (MLC) was used to analyze canthin-6-one alkaloids contained in these extracts, and the achieved performance was compared with that of a conventional high-performance liquid chromatography (HPLC) method. The optimal mobile phase of MLC corresponded to 15 : 85 (v/v) acetonitrile : water (pH 3) containing 110 mM sodium dodecyl sulfate and 10 mM NaH_2PO_4. The retention times of canthin-6-one-9-O-b-D-glucopyranoside, 9-hydroxycanthin- 6-one, canthin-6-one, and 9-methoxycanthin-6-one were 4.78/15.42, 17.64/24.11, 32.84/38.27, and 39.04/39.86 min, respectively, in the cases of isocratic MLC and conventional HPLC. In both cases, the analyte resolution exceeded 1.5. The MLC elution behavior of the examined analytes was largely determined by their hydrophobicity and ionization. The sensitivity, precision, accuracy, and per-run acetonitrile consumption of the MLC method were comparable to those of the conventional HPLC method. However, the latter method exhibited higher performance for application to EL and EH samples, particularly those with low analyte concentrations and varying sample matrices. Overall, the analysis of canthin-6-one alkaloids using MLC was limited to trace analytes due to interference by the matrix

Immunochemical Analysis of the Antimalarial Drugs Artemisinin and Artesunate

We prepared a monoclonal antibody (mAb 1C1) showing specificity for artemisinin (AM) and artesunate (AS), and we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) using this novel mAb. Moreover, we prepared a recombinant antibody derived from mAb 1C1 in order to overcome insufficient mAb production by hybridoma culture. A recombinant antigen-binding fragment (Fab) was easily constructed using antibody manipulation technologies and was produced by microorganisms in high yield. We herein review immunochemical approaches for analysis of the antimalarial drugs AM and AS that were able to yield analysis results for multiple samples in a short period of time using simple and reliable protocols

Kwakhurin-magnetic particles conjugates enable fast enzyme immunoassay for the detection of kwakhurin in Pueraria candollei

Introduction: Kwakhurin (Kwa) is one of the unique isoflavonoids produced in Pueraria candollei var. mirifica (P. candollei), which has long been used as folk medicine for rejuvenation in Thailand. Recently, the use of P. candollei-derived products has widely spread among Japanese women for cosmetic purposes. Correspondingly, there has been an increase in the number of reports regarding possible health hazards caused by estrogenic activity inherent to the plant; thus, the need for a detailed evaluation of the phytoestrogen content of P. candollei-derived products has gained a sense of urgency in recent years. / Objective: This study aims to develop a rapid enzyme immunoassay that can be applied to the quantitative analysis of Kwa in P. candollei and its derived products. / Material and Method: A rapid and sensitive immunoassay was developed with a combination of Kwa-specific monoclonal antibody (MAb 11F) and Kwa–magnetic particles (MPs) conjugates, which increased the surface area of the solid phase, resulting in a decrease in the immunoreaction time. / Result: This novel MPs-based enzyme immunoassay (MPs-EIA) was used to determine Kwa concentration in the range from 2.44 to 78.1 ng/mL with a limit of detection of 1.90 ng/mL. Validation analyses revealed that the proposed MPs-EIA protocol was sufficiently precise and accurate for effective quantitative analysis of Kwa in P. candollei and its derived products

Evaluating the <i>in Vitro</i> Efficacy of Quassinoids from <i>Eurycoma longifolia</i> and <i>Eurycoma harmandiana</i> against Common Cold Human Coronavirus OC43 and SARS-CoV‑2 Using In-Cell Enzyme-Linked Immunosorbent Assay

Coronavirus disease-2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection, has become a pandemic and public health crisis. SARS-CoV-2 and the seasonal common cold coronavirus (HCoV-OC43) belong to the beta genus of human coronaviruses (HCoVs). In-cell ELISA assays were performed using HCoV-OC43 and SARS-CoV-2 and evaluated the antiviral activity of herbal plants. Eurycoma longifolia (EL) and Eurycoma harmandiana (EH) roots (antipyretic properties) and their constituent quassinoids, especially chaparrinone and eurycomalactone, showed potent anti-HCoV-OC43 and SARS-CoV-2 activities, and the low IC50 values of the mentioned constituents were observed in the range of 0.32–0.51 μM. Eurycomanone and 13β,21-dihydroeurycomanone may contribute to the antiviral activity of EL, whereas chaparrinone is the major and active antiviral constituent of EH root. The content of quassinoids, β-carboline, and canthin-6-one alkaloids and the cytotoxicity profile of EL and EH extracts were varied regarding extraction solvents. The boiled water and 50% EtOH extractions of both plants were less toxic than those with 95% EtOH as the extraction solvent. Our research suggests that quassinoids, which come from EL and EH roots and are anti-coronavirus compounds, are potential treatment candidates for COVID-19 and merit further in vivo investigations