Analytical and Bioanalytical Chemistry

, Volume 411, Issue 12, pp 2707–2714 | Cite as

Paper spray portable mass spectrometry for screening of phorbol ester contamination in glycerol-based medical products

  • Samanthi WickramasekaraEmail author
  • Rahul Kaushal
  • Hongli Li
  • Dinesh Patwardhan
Research Paper


The Jatropha curcas plant (Jatropha) has been proposed as a source of biodiesel fuel, as it yields crude glycerol as an abundant by-product. Its by-products could serve as a starting material in making glycerol for FDA-regulated products. Jatropha is not regarded as a source of edible vegetable oil since it contains phorbol esters (PEs). PEs, even at very low exposure concentrations, demonstrate various toxicities in humans and animals, but may not be detected by routine impurity analyses. Here, we demonstrate the development of a rapid and simplified method for the detection and quantification of Jatropha-derived PE toxins using ambient ionization mass spectrometry. To do this, we successfully coupled a paper spray ambient ionization source with an ion trap portable mass spectrometer. The paper spray source was assembled using chromatography papers, and analyte ions were generated by applying a high voltage to a wetted paper triangle loaded with PE standards. For method development, we used commercially available PE standards on an ion trap portable mass spectrometer. Standard solutions were prepared using ethanol with PE concentrations ranging from 1.0 to 0.0001 mg mL−1. Spike and recovery experiments were performed using USP grade and commercially available glycerol. To discern chemical differences between samples, we applied multivariate data analysis. Based on the results obtained, paper spray coupled with a portable mass spectrometric method can be successfully adopted for the analysis of toxic contaminants present in glycerol-based consumer products with LOD and LOQ of 0.175 μg mL−1 and 0.3 μg mL−1 respectively. This direct, simple design, and low-cost sampling and ionization method enables fast screening with high sensitivity in non-laboratory settings.


Paper spray Portable mass spectrometry Phorbol esters Jatropha factors Spike and recovery Glycerol 



This project was supported in part by an appointment to the Research Participation Program at the Center for Devices and Radiological Health, administrated by the Oak Ridge Institute for Science and Education through an inter-agency agreement between US Department of Energy and the US Food and Drug Administration (FDA). Additionally, we would like to thank Dr. Steven Wolfgang from the FDA, Center for Drug Evaluation and Research, for his valuable guidance on the project, as well as Dr. Jose Centeno and Dr. Benita J. Dair, for their management support.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.


The findings and conclusions in this paper have not been formally disseminated by the FDA and should not be construed to represent any agency determination or policy. The mention of commercial products, their sources, or their use regarding material reported herein is not to be construed as either actual or implied endorsement of such products by the US Department of Health and Human Services.

Supplementary material

216_2019_1717_MOESM1_ESM.pdf (528 kb)
ESM 1 (PDF 527 kb)


  1. 1.
    Sommers CD, Keire DA. Detection of possible economically motivated adulterants in heparin sodium and low molecular weight heparins with a colorimetric microplate based assay. Anal Chem. 2011;83(18):7102–8.CrossRefGoogle Scholar
  2. 2.
    Scholl PF, Bergana MM, Yakes BJ, Xie Z, Zbylut S, Downey G, et al. Effects of the adulteration technique on the near-infrared detection of melamine in milk powder. J Agric Food Chem. 2017;65(28):5799–809.CrossRefGoogle Scholar
  3. 3.
    Ren Y, Wang H, Liu JJ, Zhang ZP, McLuckey MN, Ouyang Z. Analysis of biological samples using paper spray mass spectrometry: an investigation of impacts by the substrates, solvents and elution methods. Chromatographia. 2013;76(19–20):1339–46.CrossRefGoogle Scholar
  4. 4.
    Roux PP, Ballif BA, Anjum R, Gygi SP, Blenis J. Tumor-promoting phorbol esters and activated Ras inactivate the tuberous sclerosis tumor suppressor complex via p90 ribosomal S6 kinase. Proc Natl Acad Sci U S A. 2004;101(37):13489–94.CrossRefGoogle Scholar
  5. 5.
    Baird WM, Boutwell RK. Tumor-promoting activity of phorbol and four diesters of phorbol in mouse skin. Cancer Res. 1971;31(8):1074–9.Google Scholar
  6. 6.
    Bandomir J, Kaule S, Schmitz KP, Sternberg K, Petersen S, Kragl U. Usage of different vessel models in a flow-through cell: in vitro study of a novel coated balloon catheter. RSC Adv. 2015;5(15):11604–10.CrossRefGoogle Scholar
  7. 7.
    Sarode K, Spelber DA, Bhatt DL, Mohammad A, Prasad A, Brilakis ES, et al. Drug delivering technology for endovascular management of infrainguinal peripheral artery disease. JACC Cardiovasc Interv. 2014;7(8):827–39.CrossRefGoogle Scholar
  8. 8.
    Balaji Viswanathan RS, Kapila S, Lorbert S. Characterization and quantification of phorbol esters by tandem ESI-FT-ICR-MS. LC GC Chromatographyonlinecom. 2012;10(2):16–25.Google Scholar
  9. 9.
    Baldini M, Ferfuia C, Bortolomeazzi R, Verardo G, Pascali J, Piasentier E, et al. Determination of phorbol esters in seeds and leaves of Jatropha curcas and in animal tissue by high-performance liquid chromatography tandem mass spectrometry. Ind Crop Prod. 2014;59:268–76.CrossRefGoogle Scholar
  10. 10.
    Nishshanka U, Jayasuriya H, Chattopadhaya C, Kijak PJ, Chu P-S, Reimschuessel R, et al. Screening for toxic phorbol esters in jerky pet treat products using LC–MS. J Chromatogr B. 2016;1020:90–5.CrossRefGoogle Scholar
  11. 11.
    Herath K, Girard L, Reimschuessel R, Jayasuriya H. Application of time-of-flight mass spectrometry for screening of crude glycerins for toxic phorbol ester contaminants. J Chromatogr B. 2017;1046:226–34.CrossRefGoogle Scholar
  12. 12.
    Cooks RG, Ouyang Z, Takats Z, Wiseman JM. Ambient mass spectrometry. Science. 2006;311(5767):1566–70.CrossRefGoogle Scholar
  13. 13.
    Harris GA, Galhena AS, Fernandez FM. Ambient sampling/ionization mass spectrometry: applications and current trends. Anal Chem. 2011;83(12):4508–38.CrossRefGoogle Scholar
  14. 14.
    Liu JJ, Wang H, Manicke NE, Lin JM, Cooks RG, Ouyang Z. Development, characterization, and application of paper spray ionization. Anal Chem. 2010;82(6):2463–71.CrossRefGoogle Scholar
  15. 15.
    Wang H, Liu JJ, Cooks RG, Ouyang Z. Paper spray for direct analysis of complex mixtures using mass spectrometry. Angew Chem Int Ed. 2010;49(5):877–80.CrossRefGoogle Scholar
  16. 16.
    Michely JA, Meyer MR, Maurer HH. Paper spray ionization coupled to high resolution tandem mass spectrometry for comprehensive urine drug testing in comparison to liquid chromatography-coupled techniques after urine precipitation or dried urine spot workup. Anal Chem. 2017;89(21):11779–86.CrossRefGoogle Scholar
  17. 17.
    Wang H, Manicke NE, Yang Q, Zheng L, Shi R, Cooks RG, et al. Direct analysis of biological tissue by paper spray mass spectrometry. Anal Chem. 2011;83(4):1197–201.CrossRefGoogle Scholar
  18. 18.
    Brown H, Oktem B, Windom A, Doroshenko V, Evans-Nguyen K. Direct analysis in real time (DART) and a portable mass spectrometer for rapid identification of common and designer drugs on-site. Forensic Chem. 2016;1:66–73.CrossRefGoogle Scholar
  19. 19.
    Hua W, Hu H, Chen F, Tang L, Peng T, Wang Z. Rapid isolation and purification of phorbol esters from Jatropha curcas by high-speed countercurrent chromatography. J Agric Food Chem. 2015;63(10):2767–72.CrossRefGoogle Scholar
  20. 20.
    Roach JS, Devappa RK, Makkar HPS, Becker K. Isolation, stability and bioactivity of Jatropha curcas phorbol esters. Fitoterapia. 2012;83(3):586–92.CrossRefGoogle Scholar
  21. 21.
    Wang ZG, Tang L, Hu HL, Guo YR, Peng T, Yan F, et al. Metabolic profiling assisted quality control of phorbol esters in Jatropha curcas seed by high-performance liquid chromatography using a fused-core column. J Agric Food Chem. 2012;60(38):9567–72.CrossRefGoogle Scholar
  22. 22.
    Yang Q, Wang H, Maas JD, Chappell WJ, Manicke NE, Cooks RG, et al. Paper spray ionization devices for direct, biomedical analysis using mass spectrometry. Int J Mass Spectrom. 2012;312:201–7.CrossRefGoogle Scholar
  23. 23.
    Manicke NE, Yang Q, Wang H, Oradu S, Ouyang Z, Cooks RG. Assessment of paper spray ionization for quantitation of pharmaceuticals in blood spots. Int J Mass Spectrom. 2011;300(2):123–9.CrossRefGoogle Scholar
  24. 24.
    Espy RD, Muliadi AR, Ouyang Z, Cooks RG. Spray mechanism in paper spray ionization. Int J Mass Spectrom. 2012;325-327:167–71.CrossRefGoogle Scholar
  25. 25.
    Manicke NE, Bills BJ, Zhang C. Analysis of biofluids by paper spray MS: advances and challenges. Bioanalysis. 2016;8(6):589–606.CrossRefGoogle Scholar
  26. 26.
    Blain MG, Riter LS, Cruz D, Austin DE, Wu G, Plass WR, et al. Towards the hand-held mass spectrometer: design considerations, simulation, and fabrication of micrometer-scaled cylindrical ion traps. Int J Mass Spectrom. 2004;236(1):91–104.CrossRefGoogle Scholar
  27. 27.
    Rice JM, Dudek GO, Barber M. Mass spectra of nucleic acid derivatives. Pyrimidines J Am Chem Soc. 1965;87(20).Google Scholar
  28. 28.
    Nothias-Scaglia LF, Schmitz-Afonso I, Renucci F, Roussi F, Touboul D, Costa J, et al. Insights on profiling of phorbol, deoxyphorbol, ingenol and jatrophane diterpene esters by high performance liquid chromatography coupled to multiple stage mass spectrometry. J Chromatogr A. 2015;1422:128–39.CrossRefGoogle Scholar
  29. 29.
    Kongmany S, Hoa TT, Hanh LTN, Imamura K, Maeda Y, Boi LV. Semi-preparative HPLC separation followed by HPLC/UV and tandem mass spectrometric analysis of phorbol esters in Jatropha seed. J Chromatogr B. 2016;1038:63–72.CrossRefGoogle Scholar
  30. 30.
    Vogg G, Achatz S, Kettrup A, Sandermann H. Fast, sensitive and selective liquid chromatographic-tandem mass spectrometric determination of tumor-promoting diterpene eaters. J Chromatogr A. 1999;855(2):563–73.CrossRefGoogle Scholar
  31. 31.
    Valliyappan T, Bakhshi NN, Dalai AK. Pyrolysis of glycerol for the production of hydrogen or syn gas. Bioresour Technol. 2008;99(10):4476–83.CrossRefGoogle Scholar
  32. 32.
    Xia J, Wishart DS. Using MetaboAnalyst 3.0 for comprehensive metabolomics data analysis. Curr Protoc Bioinformatics: Wiley; 2016.Google Scholar
  33. 33.
    Xia JG, Sinelnikov IV, Han B, Wishart DS. MetaboAnalyst 3.0-making metabolomics more meaningful. Nucleic Acids Res. 2015;43(W1):W251–W7.CrossRefGoogle Scholar

Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2019

Authors and Affiliations

  1. 1.Office of Science and Engineering Laboratories, Center for Devices and Radiological HealthUS Food and Drug AdministrationSilver SpringUSA
  2. 2.Jiangsu Collaborative Innovation Center of Biomedical Functional Materials, College of Chemistry and Materials ScienceNanjing Normal UniversityNanjingChina

Personalised recommendations