Determination of Grayanotoxin-III from in Rhodendron Ponticum And Mad Honey Samples by Liquid Chromatography−Mass Spectrometry

Abstract

Grayanotoxin-III and its glycoside derivatives were isolated from their natural sources. Grayanotoxin-III was extracted from Rhododendron Ponticum L. by liquid-liquid extraction and was isolated by prep-HPLC. Grayanotoxin-III glycoside was extracted from mad honey by SPE method. For isolation of grayanotoxin-III, a reverse phase C-18 column has been used in gradient conditions starting with 40/60 % (Methanol (A) /Water (B)) following 0 min. 34/66 (A/B), 11 min. 53/47 (A/B), 19 min. 70/30 (A/B), 22 min 100/0 (A/B) at 25 oC. Isolated fragments were examined with LC-MS analysis. The main ion of grayanotoxin-III was observed at m/z 370 with 100 % abundance with some of its fragments. Grayanotoxin-III was detected at 204 nm with Diode Array Detector at 7.5 min. with the above-mentioned conditions. Grayanotoxin-III glycoside from mad honey was identified by LC-MS and NMR analysis. 

GTX-III can be transferred to honey from Rhododendron Ponticum plant according to obtained results. As a conclusion, because of the toxicity of the GTX-III compound the honey from these regions should be carefully consumed.

 

Keywords

• Grayanotoxin-III,extraction,LC-MS,Mad honey,Rhododendron Ponticum L.

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