The SpiderParacoelotes birulai: Detection and Structure Elucidation of New Acylpolyamines by On-Line Coupled HPLC-APCI-MS and HPLC-APCI-MS/MS

Abstract

The lyophilized venom of the spider Paracoelotes birulai (Araneidae: Amaurobiidae) has been analyzed. A number of acylpolyamines were found and separated. By on-line coupled high-performance liquid chromatography and atmospheric-pressure chemical-ionization mass spectrometry, the structures of the three most abundant compounds PB 490 (N-(16-guanidino-4-hydroxy-4,8,12-triazahexadecyl)-2-(4-hydroxyindol-3yl)acetamide; Fig. 3, b), PB 421 (N-(16-guanidino-4-hydroxy-4,8,12-triazahexadecyl)-2-(4-hydroxybenzamide; Fig. 4, a), and PB 448 (N-(16-amino-4-hydroxy-4,8,12-triazahexadecyl)-2-(4-hydroxyindol-3-yl)acetamide; Fig. 5,b) were elucidated. Two different types of polyamines were found in the a-palutoxins and compared with acylpolyamines from the Agelenidae spider family. The results of this investigation will initiate further chemotaxonomical studies on spiders.Introduction. ± Spider venoms attract the undivided attention of scientists interested in these unique natural mixtures of different classes of biologically active compounds. Chemists elucidate the structures of the constituents of spider venoms, biologists try to derive the mechanisms of the interactions of venoms with the spiders prey and create new specific insecticides, while pharmacists search for new drugs. The continuing discovery of new spider species also stimulates these investigations.Depending on the family and species, spider venoms contain, in diverse ratios, various natural compounds like amino acids, polyamines, purine bases, acylpolyamines, peptides, polypeptides or proteins; sometimes just some of these compounds, sometimes all of them together are present. In the last two decades, the assumption that only proteins are responsible for the toxicity of spider venoms was revised, and the activities of the compounds present in the venoms have been investigated. At the present time, several classes of natural products are believed to be the toxic constituents of spider venoms. Acylpolyamines, peptides, polypeptides, proteins, either by themselves or coacting with one another, are responsible for the toxic action of the venom.Being available in only milligram amounts, spider venoms are quite difficult to investigate by classical methods, and the successful application of NMR spectroscopy or elemental analysis for structure elucidation is more of an exception than a generally applied procedure.Paracoelotes birulai, known from several sources as Paracoelotes luctuosus

A new liquid chromatography/electrospray ionization mass spectrometry method for the analysis of underivatized aliphatic long‐chain polyamines: application to diatom‐rich sediments

“…The fragment ion observed at m/z 343.5 is likely formed by the expulsion of ammonia (17 Da) from m/z 360.4, followed by rearrangement in a ring structure, as previously observed for putrescine [23,24] and for acylpoly- amines. [25,26] All product ions observed in the MS 2 spectra of LCPA peak 6 ( Fig. 7(A)) are listed and rationalized in Table S1 (see Supporting Information).…”

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“…Recently, we have been increasingly engaged in the analysis and characterization of polyamine toxins from spiders, in particular from Agelenopsis aperta [5] and from Paracoelotes birulai [6], by on-line coupled high-performance liquid chromatography and atmospheric-pressure chemical-ionization mass spectrometry (HPLC-UV(DAD)-APCI-MS and MS/MS). In the venom of A. aperta, e.g., as many as 33 acylpolyamines with 11 different molecular masses were detected by this method.…”

The Acylpolyamines from the Venom of the SpiderAgelenopsis aperta

“…Last year, high-performance liquid chromatography (HPLC), coupled on-line with atmospheric-pressure chemical-ionization mass spectroscopy (APCI-MS), has been successfully used for the isolation and structure elucidation of polyamines from the venom of the spider Paracoelotes birulai (Aranea: Amaurobiidae) [22]. It gave an idea that this selective and sensitive method could be used for analysis and structure elucidation of new acylpolyamines present in Agelenopsis aperta venom in low concentrations.…”

“…This sensitive and selective approach, which was already efficiently applied for the structure elucidation of acylpolyamines present in the Paracoelotes birulai [22] spider venom, can be employed for the characterization of new compounds present in low concentrations as well as to those, co-eluted with another one.…”

“…This investigation allowed the structure determination of polyamine backbones. The position of the OH group either at the chromophore or at the N-atoms was deduced as for acylpolyamines from the Paracoelotes birulai spider venom [22].…”