Stability and epimerisation behaviour of ergot alkaloids in various solvents

Abstract

In this paper the stability and degree of epimerisation of six major ergot alkaloids at three different temperature levels (-20 °C, +4 °C and +20 °C) over periods of 18 hours and six weeks is reported for the first time. The behaviour of ergometrine, ergocornine, ergocristine, α-ergocryptine, ergosine and ergotamine was thoroughly studied in seven solvents which are employed for the preparation of calibrants and extraction mixtures, respectively. Moreover, the stability of the ergot alkaloids was tested in different cereal extracts (rye, wheat, barley, oats) for 1, 2 and 6 days. Of the toxins tested, the ergopeptide-type toxins ergosine, ergotamine, ergocornine, α-ergocryptine and ergocristine showed similar behaviour patterns. The simple lysergic acid derivative ergometrine was more stable and showed hardly any epimerisation to ergometrinine, with the sum of both epimers remaining constant in all seven solvents. The ergopeptides tested show variable epimerisation tendencies, and were also less stable during six weeks at 20 °C. Ergosine showed the highest degree of epimerisation (43% after 6 weeks at 20 °C). In general, the order of epimerisation promotion was methanol/dichloromethane > acetonitrile/buffer > extraction mix > stabilising solution > acetonitrile >> chloroform. Long-term storage at room temperature can only be carried out in chloroform, which showed no epimerisation for all toxins even at 20 °C and also kept the sum of R and S forms constant, which indicates no formation of aci-epimers or other degradation products. Long-term storage of ergot alkaloids in acetonitrile, the most convenient solvent with respect to HPLC analysis, should be carried out at temperatures of -20 °C or below. The constant epimer ratio of all ergot alkaloids in the extraction mixture acetonitrile/ammonium carbonate buffer (200 mg/l; 92:8, v/v) during an HPLC run (18 hours) demonstrates the stability of the toxins in this extraction mixture.