Baiting Insects with Pyrrolizidine Alkaloids (PAs): A Fieldwork-Oriented Review and Guide to PA-Pharmacophagy.

Since 1890, many observations of danaine butterflies visiting dry plants of several families in the Old and New World tropics have been published. For 50 years, it has been recognised that Danainae, along with various other insects, seek out 1,2-dehydropyrrolizidine ester alkaloids (PAs) independently of and in addition to their nutritive requirements and utilise them to increase their chances for survival and biological fitness. This represents an unusual type of insect-plant relationship ("PA-pharmacophagy"), with remarkable peculiarities but also with gaps in knowledge, many of which can be filled by employing PA-baiting. We review and analyse the history of records on the attraction of adult insects to PAs and unveil the complex background information on PA-chemistry, PA-producing plants ("PA-plants"), and PA-sequestering insects ("PA-insects") in order to come up with practical tips for successful baiting with PAs ("PA-baiting"). Studying PA-pharmacophagy integrates taxonomy, behaviour, and ecology with evolutionary biology, chemistry, and toxicology. With basic knowledge of PA-chemistry and bearing the general peculiarities of PA-plants and PA-insects in mind, PA-baiting can be conducted easily and successfully to address many questions on the multifaceted ecology of pyrrolizidine alkaloids. We aim to encourage field researchers in the tropics to employ PA-baiting as a valuable research method in this field of integrative biology.

Pro-toxic dehydropyrrolizidine alkaloids in the traditional Andean herbal medicine "asmachilca".

ETHNOPHARMACOLOGICAL RELEVANCE: Asmachilca is a Peruvian medicinal herb preparation ostensibly derived from Aristeguietia gayana (Wedd.) R.M. King & H. Rob. (Asteraceae: Eupatorieae). Decoctions of the plant have a reported bronchodilation effect that is purported to be useful in the treatment of respiratory allergies, common cold and bronchial asthma. However, its attractiveness to pyrrolizidine alkaloid-pharmacophagous insects indicated a potential for toxicity for human consumers. AIM OF THE STUDY: To determine if commercial asmachilca samples, including fully processed herbal teas, contain potentially toxic 1,2-dehydropyrrolizidine alkaloids. MATERIALS AND METHODS: Two brands of "Asmachilca" herbal tea bags and four other commercial samples of botanical materials for preparing asmachilca medicine were extracted and analyzed using HPLC-esi(+)MS and MS/MS for the characteristic retention times and mass spectra of known dehydropyrrolizidine alkaloids. Other suspected dehydropyrrolizidine alkaloids were tentatively identified based on MS/MS profiles and high resolution molecular weight determinations. Further structure elucidation of isolated alkaloids was based on 1D and 2D NMR spectroscopy. RESULTS: Asmachilca attracted many species of moths which are known to pharmacophagously gather dehydropyrrolizidine alkaloids. Analysis of 5 of the asmachilca samples revealed the major presence of the dehydropyrrolizidine alkaloid monoesters rinderine and supinine, and their N-oxides. The 6th sample was very similar but did not contain supinine or its N-oxide. Small quantities of other dehydropyrrolizidine alkaloid monoesters, including echinatine and intermedine, were also detected. In addition, two major metabolites, previously undescribed, were isolated and identified as dehydropyrrolizidine alkaloid monoesters with two "head-to-tail" linked viridifloric and/or trachelanthic acids. Estimates of total pyrrolizidine alkaloid and N-oxide content in the botanical components of asmachilca varied from 0.4% to 0.9% (w/dw, dry weight) based on equivalents of lycopsamine. The mean pyrrolizidine alkaloid content of a hot water infusion of a commercial asmachilca herbal tea bag was 2.2±0.5mg lycopsamine equivalents. Morphological and chemical evidence showed that asmachilca is prepared from different plant species. CONCLUSIONS: All asmachilca samples and the herbal tea infusions contained toxicologically-relevant concentrations of pro-toxic 1,2-dehydropyrrolizidine alkaloid esters and, therefore, present a risk to the health of humans. This raises questions concerning the ongoing unrestricted availability of such products on the Peruvian and international market. In addition to medical surveys of consumers of asmachilca, in the context of chronic disease potentially associated with ingestion of the dehydropyrrolizidine alkaloids, the botanical origins of asmachilca preparations require detailed elucidation.