Abstract
Pyrrolizidine alkaloids (PAs) and their N-oxides (PANOs) can cause liver toxicity and genotoxicity in humans and animals, necessitating the development of screening tools for these alkaloids. Ultra-performance liquid chromatography–electrospray ionization-quadrupole-time-of-flight mass spectrometry (UPLC–ESI–Q-TOFMS) can provide accurate precursor and fragment ion mass information (MSE mode) as well as simultaneous quantification. A method for rapidly and accurately determining nine PAs with UPLC–ESI–Q-TOF MS was developed and validated herein. Analytes were separated using a C18 column with water containing 0.1% formic acid and acetonitrile, showing high-resolution peaks within 11 min. The validation results showed excellent linearity (R2 > 0.9951), limits of detection (0.4–2.0 ng mL−1), and limits of quantification (0.6–6.0 ng mL−1). Moreover, 70 PAs, their N-oxides precursors, and characteristic fragment ions that were generated according to their chemical structure were characterized. A method for the chemical profiling of alkaloids was also proposed using the mass information obtained from the chromatograms of the tested sample. Lycopsamine, senecionine, senkirkine, and echimidine were identified in four potentially PA-containing plants and quantified by matching with authentic standards. Eight PAs and PANOs were also tentatively identified using the mass data from the previously listed alkaloids. This approach will provide a database that can be used to instantly identify alkaloids in botanical samples.
Similar content being viewed by others
References
Petruczynik A (2012) Analysis of alkaloids from different chemical groups by different liquid chromatography methods. Cent Eur J Chem 10(3):802–835
He X, Xia Q, Woodling K, Lin G, Fu PP (2017) Pyrrolizidine alkaloid-derived DNA adducts are common toxicological biomarkers of pyrrolizidine alkaloid N-oxides. J Food Drug Anal 25(4):984–991
Wiedenfeld H (2013) Alkaloids derived from ornithine: pyrrolizidine alkaloids. In: Ramawat K, Mérillon JM (eds) Natural Products. Springer, Berlin, Heidelberg, pp 359–379
Kempf M, Reinhard A, Beuerle T (2010) Pyrrolizidine alkaloids (PAs) in honey and pollen-legal regulation of PA levels in food and animal feed required. Mol Nutr Food Res 54(1):158–168
Zhu L, Ruan JQ, Li N, Fu PP, Ye Y, Lin G (2016) A novel ultra-performance liquid chromatography hyphenated with quadrupole time of flight mass spectrometry method for rapid estimation of total toxic retronecine-type of pyrrolizidine alkaloids in herbs without requiring corresponding standards. Food Chem 194:1320–1328
Letsyo E, Jerz G, Winterhalter P, Beuerle T (2017) Toxic pyrrolizidine alkaloids in herbal medicines commonly used in Ghana. J Ethnopharmacol 202:154–161
Martinello M, Borin A, Stella R, Bovo D, Biancotto G, Gallina A, Mutinelli F (2017) Development and validation of a QuEChERS method coupled to liquid chromatography and high resolution mass spectrometry to determine pyrrolizidine and tropane alkaloids in honey. Food Chem 234:295–302
Schaneberg BT, Molyneux RJ, Khan IA (2004) Evaporative light scattering detection of pyrrolizidine alkaloids. Phytochem Anal PCA 15(1):36–39
Crews C, Startin JR, Clarke PA (1997) Determination of pyrrolizidine alkaloids in honey from selected sites by solid phase extraction and HPLC–MS. Food Addit Contam 14(5):419–428
Zhou Y, Li N, Choi FF, Qiao CF, Song JZ, Li SL, Liu X, Cai ZW, Fu PP, Lin G, Xu HX (2010) A new approach for simultaneous screening and quantification of toxic pyrrolizidine alkaloids in some potential pyrrolizidine alkaloid-containing plants by using ultra performance liquid chromatography–tandem quadrupole mass spectrometry. Anal Chim Acta 681(1–2):33–40
Schurmann A, Dvorak V, Cruzer C, Butcher P, Kaufmann A (2009) False-positive liquid chromatography/tandem mass spectrometric confirmation of sebuthylazine residues using the identification points system according to EU directive 2002/657/EC due to a biogenic insecticide in tarragon. Rapid Commun Mass Spectrom RCM 23(8):1196–1200
Xie S, Shi Y, Wang Y, Wu C, Liu W, Feng F, Xie N (2013) Systematic identification and quantification of tetracyclic monoterpenoid oxindole alkaloids in Uncaria rhynchophylla and their fragmentations in Q-TOF–MS spectra. J Pharm Biomed Anal 81–82:56–64
Jeong WT, Lim HB (2018) A UPLC–ESI–Q-TOF method for rapid and reliable identification and quantification of major indole alkaloids in Catharanthus roseus. J Chromatogr B Anal Technol Biomed Life Sci 1080:27–36
Nordstrom A, O’Maille G, Qin C, Siuzdak G (2006) Nonlinear data alignment for UPLC–MS and HPLC–MS based metabolomics: quantitative analysis of endogenous and exogenous metabolites in human serum. Anal Chem 78(10):3289–3295
Le PM, McCooeye M, Windust A (2014) Application of UPLC–QTOF–MS in MS(E) mode for the rapid and precise identification of alkaloids in goldenseal (Hydrastis canadensis). Anal Bioanal Chem 406(6):1739–1749
Qi Y, Li S, Pi Z, Song F, Lin N, Liu S, Liu Z (2014) Chemical profiling of Wu-tou decoction by UPLC–Q-TOF–MS. Talanta 118:21–29
Avula B, Sagi S, Wang YH, Zweigenbaum J, Wang M, Khan IA (2015) Characterization and screening of pyrrolizidine alkaloids and N-oxides from botanicals and dietary supplements using UHPLC-high resolution mass spectrometry. Food Chem 178:136–148
Picron JF, Herman M, Van Hoeck E, Goscinny S (2018) Analytical strategies for the determination of pyrrolizidine alkaloids in plant based food and examination of the transfer rate during the infusion process. Food Chem 266:514–523
Bolechova M, Caslavsky J, Pospichalova M, Kosubova P (2015) UPLC–MS/MS method for determination of selected pyrrolizidine alkaloids in feed. Food Chem 170:265–270
Perez-Arribas LV, Manuel de Villena-Rueda FJ, Leon-Gonzalez ME, Gonzalo-Lumbreras R, Polo-Diez LM (2010) New approach to optimize HPLC separations of acid-base compounds with elution order involved, by using combined three-band resolution maps. Anal Bioanal Chem 396(7):2647–2656
Buckenmaier SM, McCalley DV, Euerby MR (2002) Overloading study of bases using polymeric RP-HPLC columns as an aid to rationalization of overloading on silica-ODS phases. Anal Chem 74(18):4672–4681
Lin G, Zhou KY, Zhao XG, Wang ZT, But PP (1998) Determination of hepatotoxic pyrrolizidine alkaloids by on-line high performance liquid chromatography mass spectrometry with an electrospray interface. Rapid Commun Mass Spectrom RCM 12(20):1445–1456
van de Schans MG, Blokland MH, Zoontjes PW, Mulder PP, Nielen MW (2017) Multiple heart-cutting two dimensional liquid chromatography quadrupole time-of-flight mass spectrometry of pyrrolizidine alkaloids. J Chromatogr A 1503:38–48
Kempf M, Wittig M, Reinhard A, von der Ohe K, Blacquière T, Raezke KP, Michel R, Schereier P, Beuerle T (2011) Pyrrolizidine alkaloids in honey: comparison of analytical methods. Food Addit Contam Part A 28(3):332–347
Colegate SM, Gardner DR, Joy RJ, Betz JM, Panter KE (2012) Dehydropyrrolizidine alkaloids, including monoesters with an unusual esterifying acid, from cultivated Crotalaria juncea (Sunn Hemp cv. ‘Tropic Sun’). J Agric Food Chem 60(14):3541–3550
Colegate SM, Welsh SL, Gardner DR, Betz JM, Panter KE (2014) Profiling of dehydropyrrolizidine alkaloids and their N-oxides in herbarium-preserved specimens of Amsinckia species using HPLC-esi (+) MS. J Agric Food Chem 62(30):7382–7392
Mudge EM, Jones AMP, Brown PN (2015) Quantification of pyrrolizidine alkaloids in North American plants and honey by LC–MS: single laboratory validation. Food Addit Contam Part A 32(12):2068–2074
Wuilloud JC, Gratz SR, Gamble BM, Wolnik KA (2004) Simultaneous analysis of hepatotoxic pyrrolizidine alkaloids and N-oxides in comfrey root by LC-ion trap mass spectrometry. Analyst 129(2):150–156
Lebada R, Schreier A, Scherz S, Resch C, Krenn L, Kopp B (2000) Quantitative analysis of the pyrrolizidine alkaloids senkirkine and senecionine in Tussilago farfara L. by capillary electrophoresis. Phytochem Anal 11(6):366–369
Liu F, Wan SY, Jiang Z, Li SF, Ong ES, Osorio JC (2009) Determination of pyrrolizidine alkaloids in comfrey by liquid chromatography–electrospray ionization mass spectrometry. Talanta 80(2):916–923
Funding
No funding was provided for this study.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare there are no conflicts of interest regarding the publication of this paper.
Ethical approval
This article does not contain any studies with human participants or animals performed by authors. The authors confirm compliance with ethical standards.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Jeong, W.T., Lim, H.B. Determination and Chemical Profiling of Toxic Pyrrolizidine Alkaloids in Botanical Samples with UPLC–Q-TOFMS. Chromatographia 82, 1653–1664 (2019). https://doi.org/10.1007/s10337-019-03785-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10337-019-03785-y