Determination of residual levels of naloxone, yohimbine, thiophanate, and altrenogest in porcine muscle using QuEChERS with liquid chromatography and triple quadrupole mass spectrometry.

A quick, easy, cheap, effective, rugged, and safe QuEChERS (method) was used for the simultaneous detection of four veterinary drug residues, namely naloxone, yohimbine, thiophanate, and altrenogest, in porcine muscle, using liquid chromatography with electrospray ionization triple quadrupole tandem mass spectrometry. Because of the unavailability of a suitable internal standard, matrix-matched calibrations were used for quantification, with determination coefficients ≥ 0.9542. The accuracy (expressed as recovery %) ranged from 60.53 to 83.25%, and the intra- and interday precisions (expressed as relative standard deviations) were <12%. The limits of quantification were 5, 0.5, 2, and 5 ng/g for naloxone, yohimbine, thiophanate, and altrenogest, respectively. Samples purchased from local markets in Seoul, Republic of Korea, revealed no traces of the target analytes. The developed method described herein is sensitive and reliable and can be applied to quantify the tested veterinary drugs in animal tissues.

Determination of terpenoid indole alkaloids in hairy roots of Rhazya stricta (Apocynaceae) by GC-MS.

INTRODUCTION: Rhazya stricta Decne. (Apocynaceae) is a medicinal plant rich in terpenoid indole alkaloids (TIAs), some of which possess important pharmacological properties. The study material including transgenic hairy root cultures have been developed and their potential for alkaloid production are being investigated. OBJECTIVE: In this study, a comprehensive GC-MS method for qualitative and quantitative analysis of alkaloids from Rhazya hairy roots was developed. METHODS: The composition of alkaloids was determined by using GC-MS. In quantification, the ratio between alkaloid and internal standard was based on extracted ion from total ion current (TIC) analyses. RESULTS: The developed method was validated. An acceptable precision with RSD ≤ 8% over a linear range of 1 to 100 µg/mL was achieved. The accuracy of the method was within 94-107%. Analysis of hairy root extracts indicated the occurrence of a total of 20 TIAs. Six of them, pleiocarpamine, fluorocarpamine, vincamine, ajmalicine and two yohimbine isomers are reported here for the first time in Rhazya. Trimethylsilyl (TMS) derivatisation of the extracts resulted in the separation of two isomers for yohimbine and also for vallesiachotamine. Clearly improved chromatographic profiles of TMS-derivatives were observed for vincanine and for minor compounds vincamine and rhazine. CONCLUSION: The results show that the present GC-MS method is reliable and well applicable for studying the variation of indole alkaloids in Rhazya samples.

Rauniticine-allo-oxindole B and rauniticinic-allo acid B, new heteroyohimbine-type oxindole alkaloids from the stems of Malaysian Uncaria longiflora var. pteropoda.

Two new heteroyohimbine-type oxindole alkaloids, rauniticine-allo-oxindole B and rauniticinic-allo acid B, have been successfully isolated from the stems extract of Malaysian Uncaria longiflora var. pteropoda. The structures of the two new alkaloids were determined by spectroscopic analysis.

Plant-cell bioreactors with simultaneous electropermeabilization and electrophoresis.

Experimental investigations on using low-level electric currents and voltages to extract, transport, and collect intracellular secondary metabolites from plant cells while maintaining their viabilities were conducted focusing on the production of: (1) ionic betalains, mainly negatively-charged betanin, from Beta vulgaris cells, and (2) ionic alkaloids, particularly positively-charged ajmalicine and yohimbine, from Catharanthus roseus cells. Three versions of tubular membrane reactors in which electropermeabilization of cell membranes and electrophoresis and diffusion of ionic products take place simultaneously, with or without convective flow, to achieve desirable extraction were developed. Concentrations of secondary metabolites produced from these plant-cell reactors under steady and oscillatory electrical forcings were recorded and the viabilities of treated cells examined. Oscillatory application of electrical field appears to produce more products while retaining higher cell viability.

Studies on production of ajmalicine in shake flasks by multiple shoot cultures of Catharanthus roseus.

The effects of different concentrations of indole-3-acetic acid (IAA) and benzyladenine (BA) on production of ajmalicine by multiple shoot cultures of Catharanthus roseus (C. roseus) were studied. By supplementing Murashige and Skoog's (MS) medium with a high concentration of IAA (11.42 microM) and a low concentration of BA (2.22 microM), shoot cultures accumulated high levels of ajmalicine. When culture medium was fortified with a low concentration of IAA (2.85 microM) and a high concentration of BA (8.90 microM), shoots released high levels of ajmalicine into the culture medium. Quantification of ajmalicine was performed by high performance liquid chromatography (HPLC). The highest concentration of ajmalicine production (0.166% dry wt) was obtained by shoot cultures grown in MS medium containing IAA (11.42 microM) on 20 days of cultivation. Shoot cultures accumulated ajmalicine 4.2-fold more in IAA (11.42 microM) supplemented medium compared with the high concentration of BA (8.90 microM). The content of ajmalicine concentration in the medium was quantified. Shoot cultures grown in BA (8.90 microM) supplemented medium released the maximum production of ajmalicine (0.853 g/L) into the culture medium after 15 days of cultivation. The experimental data show that the secretion of ajmalicine was 2-fold more into the culture medium supplemented with a high concentration of BA compared to that with a low concentration of BA. Data presented here show that production of ajmalicine by shoot cultures is not correlated with growth rate. Dimeric indole alkaloids vincristine and vinblastine were not present in shoot cultures. Ajmalicine production by shoot cultures was 2.4-fold higher compared to leaves of 1-year-old naturally grown plants.

Catharanthus alkaloids XXXII: isolation of alkaloids from Catharanthus trichophyllus roots and structure elucidation of cathaphylline.

Further examination of the cytotoxic alkaloid fractions of Catharanthus trichophyllus roots afforded nine alkaloids. Two of these alkaloids, lochnericine and horhammericine, are responsible for part of the cytotoxic activity. The structure elucidation of cathaphylline, a new beta-anilino acrylate derivative, is described.

Isolation and biological activity of aspidospermine and quebrachamine from an Aspidosperma tree source.

The indolealkaloids aspidospermine and quebrachamine have been isolated in crystalline form by a relatively rapid fractionation from the extract of a powdered material designated "Quebracho" derived from an Aspidosperma tree species. We present a novel isocratic LC method that provides baseline resolution of these two compounds and of the structurally related yohimbine in less than 15 min. Gas chromatography-mass spectrometry was employed to identify these compounds as well as several minor derivatives of aspidospermine during and after the purification process. Aspidospermine and quebrachamine like yohimbine have been found to possess adrenergic blocking activities for a variety of urogenital tissues.

[Studies on the chemical constituents of Uncaria yunanensis Hsia. C.C].

AIM: To research the chemical constituents from dried roots of Uncaria yunanensis Hsia. C. C. METHODS: Modern chromatography was used to isolate chemical components. Their structure were identified by spectral analysis. RESULTS: Seven compounds were isolated and identified as 3 beta, 6 beta, 19 alpha-trihydroxyurs-12-en-28 oic acid (I), 23-nor-24-esomethylene-3 beta, 6 beta-19 alpha-trihydroxyurs-12-en-28 oic acid (II), 3-oxo-6 beta, 19 alpha-dihydroxyurs-12-en-28 oic acid (III), oleanic acid (IV), 5,7,3',4'-tetrahydroxy-flavan-3-ol (V), beta-yohimbine (VI) and diangoutengjian I (VII). CONCLUSION: All of the above compounds were isolated for the first time from the root of this plant. Among them, compound VII is a new one.

[Separation and preparation of indole alkaloids in Lycorma delicatula White. by HPLC].

A HPLC method for separating and preparing indole alkaloids is described. HPLC conditions for analysis: BIO-RAD series 700 HPLC, model 700 data station, UV: model 1749 UV-VIS monitor, column: BIO-RAD Hi-pore RP318, 250 mm x 10 mm, mobile phase: 80% methanol-H2O(gradient), flow rate: 1.5 ml/min, detection wavelength: 254 nm. On the basis of spectral (1HNMR, 13CNMR, H-H COSY, MS, DEPT) and chemical evidence, the structures of two compounds were elucidated as beta-yohimbine (yohimban-16-carboxylic acid-17-hydroxy methylester (3 alpha, 16 alpha, 17 beta)) and ajmalicine (oxayohimban-16-carboxylic acid-16,17-didehydro-19-ethyl methyl ester (19 alpha)).

Suspension culture of Catharanthus roseus crown gall cell induced by Agrobacterium C58.

In comparison with calli from leaf or stem, Catharanthus roseus crown gall cell cultured on MS basic medium was superior in growth, total indole alkaloids and ajmalicine contents. The effects of illumination, cultural temperature, sucrose level of the medium and exogenous L-Trp on the growth, total indole alkaloids and ajmalicine contents of C. roseus crown gall cell cultures were studied. The results will provide a theoretical basis for the attempt of using suspension cultures of C. roseus crown gall cells to produce indole alkaloids.

A new 9-methoxyyohimbine-type indole alkaloid from Mitragyna africanus.

A new yohimbine-type indole alkaloid (1). was isolated from the stem bark of Mitragyna africanus (WILLD.) collected in Nigeria, along with known seven Corynanthe-type oxindole alkaloids, two secoiridoids, three lignans, and a quinovic acid derivative. Their structures were elucidated by spectroscopic analyses.