Analysis of Pyrrolizidine Alkaloids in Queensland Honey: Using Low Temperature Chromatography to Resolve Stereoisomers and Identify Botanical Sources by UHPLC-MS/MS

Abstract

Pyrrolizidine alkaloids (PAs) are a diverse group of plant secondary metabolites with known varied toxicity. Consumption of 1,2-unsaturated PAs has been linked to acute and chronic liver damage, carcinogenicity and death, in livestock and humans, making their presence in food of concern to food regulators in Australia and internationally. In this survey, honey samples sourced from markets and shops in Queensland (Australia), were analysed by high-resolution Orbitrap UHPLC-MS/MS for 30 common PAs. Relationships between the occurrence of pyrrolizidine alkaloids and the botanical origin of the honey are essential as pyrrolizidine alkaloid contamination at up to 3300 ng/g were detected. In this study, the predominant alkaloids detected were isomeric PAs, lycopsamine, indicine and intermedine, exhibiting identical MS/MS spectra, along with lesser amounts of each of their N-oxides. Crucially, chromatographic UHPLC conditions were optimised by operation at low temperature (5 • C) to resolve these key isomeric PAs. Such separation of these isomers by UHPLC, enabled the relative proportions of these PAs present in honey to be compared to alkaloid levels in suspect source plants. Overall plant pyrrolizidine alkaloid profiles were compared to those found in honey samples to help identify the most important plants responsible for honey contamination. The native Australian vines of Parsonsia spp. are proposed as a likely contributor to high levels of lycopsamine in many of the honeys surveyed. Botanical origin information such as this, gained via low temperature chromatographic resolution of isomeric PAs, will be very valuable in identifying region of origin for honey samples. Key Contribution:This study demonstrated the occurrence of indicine and the isomeric lycopsamine as predominant pyrrolizidine alkaloids in Australian honey; and established a HRAM LC-MS/MS method that chromatographically separated these stereoisomers; enabling the investigation of botanical origin of honey contamination by the respective alkaloids.

Mineral and Trace Element Analysis of Australian/Queensland Apis mellifera Honey

“…Honey purity and origin exerts a significant influence on the commercial value of the product and is of utmost importance to consumers of Australian honey. There are few studies of the composition of Australian honey [ 13 , 27 , 28 , 29 ]. The levels identified herein constitute a first Australian study to identify mineral levels in honey and compare honey from urban versus peri-urban versus rural settings, with blended honeys.…”

Haematological and serum biochemical reference intervals of free‐ranging Lumholtz's tree‐kangaroos (Dendrolagus lumholtzi)

“…12 Examples include: phenolic compounds in privet (Ligustrum species) 13 ; alkaloids, terpenes, lignans and essential oils in laurel (Litsea species) 14 ; saponins and triterpenes in sarsaparilla (Alphitonia spp.) 15 ; pyrrolizidine alkaloids in silk pod vine (Parsonsia straminea) 16 ; alkaloids and glycosides in lantana (Lantana camara) 17 ; terpenes and essential oils in camphor laurel (Cinnamomum camphora) 18 and alkaloids and glycosides in tobacco bush (Solanum mauritianum). 19 Hepatic microsomal enzymes enable generalized detoxification pathways and specifically oxidation, reduction, hydrolysis, esterification, N-dealkylation, and conjugation of toxicants.…”

Chemotaxonomic Investigation of Apocynaceae for Retronecine-Type Pyrrolizidine Alkaloids Using HPLC-MS/MS

“…In Parsonia straminea (R.Br.) F.Muell., 96% of PAs identified via high resolution accurate mass (HRAM) LC-MS/MS in leaves and stems were N-oxides (Hungerford et al, 2019). Therefore, not The PREC scans employed in this study were designed to specifically target fragments inherent to pro-toxic, free base, 1,2-unsaturated retronecine-type PAs.…”