Advertisement

Particle Size Characterization of Sepia Ink Eumelanin Biopolymers by SEM, DLS, and AF4-MALLS: a Comparative Study

  • Inmaculada de la CalleEmail author
  • Diego Soto-Gómez
  • Paula Pérez-Rodríguez
  • J. Eugenio López-Periago
Article
  • 17 Downloads

Abstract

Cephalopod ink is a complex mixture of bioactive substances with technical properties of interest in many fields (e.g., biophysics, ecology, environment, biomedicine, food technology, cosmetics, or fine arts). It was previously reported that organic nanoparticles may naturally appear in this mixture. Thus, the particle size determination of these biopolymers is interesting from the point of view of food nanotechnology and nanotoxicology. In this work, the particle size of purified eumelanin microspheres from commercial sepia ink was successfully measured by three techniques: Scanning electron microscopy (SEM), dynamic light scattering (DLS), and asymmetric-flow field-flow fractionation linked to multi-angle laser-light scattering (AF4-MALLS). This study shows the potential and differences of the application of these techniques in terms of sample preparation, conditioning, introduction, and principles for particle size characterization of natural organic nanoparticles in foods. Thus, this methodology can be a model for the characterization of other natural and engineered organic nanoparticles in this matrix type. DLS and AF4-MALLS provide the size corresponding to the hydrodynamic diameter, which is usually larger than the size of the dense core provided by SEM (without hydration or solvation layer). Additionally, SEM informs about the particles morphology, showing a quasi-spherical shape for particles between 100 and 140 nm. DLS and AF4-MALLS indicate particles of hydrodynamic diameter in the range of 180–260 nm. Furthermore, the absolute molar mass of particles has been measured by MALLS.

Graphical Abstract

Keywords

Sepia ink DLS AF4-MALLS SEM Eumelanin Particle size 

Notes

Acknowledgements

The authors thank the Ultra Trace Analysis Aquitaine UT2A/ADERA (Pau, France) for the analysis performed on DLS and AF4-MALLS and the CACTI services from Universidade de Vigo for the SEM photographs (Vigo, Spain).

Funding Information

I. De la Calle thanks Xunta de Galicia for financial support as a postdoctoral researcher of the I2C program (POS-B/2017/012-PR) and co-financed by the European Social Funding P.P. 0000 421S 140.08. D. Soto-Gómez is funded by a predoctoral Fellowship Program (FPU) from the Spanish Ministry of Education. P. Pérez-Rodríguez is funded by a postdoctoral contract from Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (ED481B 2017/31).

Compliance with Ethical Standards

Conflict of Interest

Inmaculada de la Calle declares that she has no conflict of interest. Diego Soto-Gómez declares that he has no conflict of interest. Paula Pérez-Rodríguez declares that she has no conflict of interest. J. Eugenio López-Periago declares that he has no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Informed consent is not applicable for this study.

Supplementary material

12161_2019_1448_MOESM1_ESM.docx (1.5 mb)
ESM 1 (DOCX 1540 kb)

References

  1. Al-Saidi HM (2016) Biosorption using chitosan thiourea polymer as an extraction and preconcentration technique for copper prior to its determination in environmental and food samples by flame atomic absorption spectrometry: synthesis, characterization and analytical applications. Int J Biol Macromol 93:390–401.  https://doi.org/10.1016/j.ijbiomac.2016.08.060 Google Scholar
  2. Baalousha M, Kammer FVD, Motelica-Heino M, Hilal HS, le Coustumer P (2006) Size fractionation and characterization of natural colloids by flow-field flow fractionation coupled to multi-angle laser light scattering. J Chromatogr A 1104:272–281.  https://doi.org/10.1016/j.chroma.2005.11.095 Google Scholar
  3. Baalousha M, Kammer FVD, Motelica-Heino M, Le Coustumer P (2005) Natural sample fractionation by FlFFF-MALLS-TEM: sample stabilization, preparation, pre-concentration and fractionation. J Chromatogr A 1093:156–166.  https://doi.org/10.1016/j.chroma.2005.07.103 Google Scholar
  4. Baalousha M, Stolpe B, Lead JR (2011) Flow field-flow fractionation for the analysis and characterization of natural colloids and manufactured nanoparticles in environmental systems: a critical review. J Chromatogr A 1218:4078–4103.  https://doi.org/10.1016/j.chroma.2011.04.063 Google Scholar
  5. Crippa R, Horak V, Prota G et al (1990) Chapter 6 chemistry of melanins. In: Brossi A (ed) The alkaloids: chemistry and pharmacology. Academic Press, pp 253–323Google Scholar
  6. Derby CD (2014) Cephalopod ink: production, chemistry, functions and applications. Mar Drugs 12:2700–2730.  https://doi.org/10.3390/md12052700 Google Scholar
  7. Dubascoux S, Von Der Kammer F, Le Hécho I et al (2008) Optimisation of asymmetrical flow field flow fractionation for environmental nanoparticles separation. J Chromatogr A 1206:160–165.  https://doi.org/10.1016/j.chroma.2008.07.032 Google Scholar
  8. Ehara K, Takahata K, Koike M (2006) Absolute mass and size measurement of monodisperse particles using a modified Millikan’s method: part II—application of electro-gravitational aerosol balance to polystyrene latex particles of 100 nm to 1 μm in average diameter. Aerosol Sci Technol 40:521–535.  https://doi.org/10.1080/02786820600714387 Google Scholar
  9. Geiss O, Cascio C, Gilliland D, Franchini F, Barrero-Moreno J (2013) Size and mass determination of silver nanoparticles in an aqueous matrix using asymmetric flow field flow fractionation coupled to inductively coupled plasma mass spectrometer and ultraviolet-visible detectors. J Chromatogr A 1321:100–108.  https://doi.org/10.1016/j.chroma.2013.10.060 Google Scholar
  10. Gorniak T, Haraszti T, Suhonen H, Yang Y, Hedberg-Buenz A, Koehn D, Heine R, Grunze M, Rosenhahn A, Anderson MG (2014) Support and challenges to the melanosomal casing model based on nanoscale distribution of metals within iris melanosomes detected by X-ray fluorescence analysis. Pigment Cell Melanoma Res 27:831–834.  https://doi.org/10.1111/pcmr.12278 Google Scholar
  11. Hupfeld S, Ausbacher D, Brandl M (2009) Asymmetric flow field-flow fractionation of liposomes: optimization of fractionation variables. J Sep Sci 32:1465–1470.  https://doi.org/10.1002/jssc.200800626 Google Scholar
  12. Jarzębski M, Bellich B, Białopiotrowicz T, Śliwa T, Kościński J, Cesàro A (2017) Particle tracking analysis in food and hydrocolloids investigations. Food Hydrocoll 68:90–101.  https://doi.org/10.1016/j.foodhyd.2016.09.037 Google Scholar
  13. Lang T, Eslahian KA, Maskos M (2012) Ion effects in field-flow fractionation of aqueous colloidal polystyrene - Lang - 2012 - macromolecular chemistry and physics - Wiley online library. Macromol Chem Phys 213:2353–2361Google Scholar
  14. Liu Y, Simon JD (2003) The effect of preparation procedures on the morphology of melanin from the ink sac of sepia officinalis. Pigment Cell Res 16:72–80.  https://doi.org/10.1034/j.1600-0749.2003.00009.x Google Scholar
  15. Loiseleux T, Rolland-Sabaté A, Garnier C, Croguennec T, Guilois S, Anton M, Riaublanc A (2018) Determination of hydro-colloidal characteristics of milk protein aggregates using asymmetrical flow field-flow fractionation coupled with multiangle laser light scattering and differential refractometer (AF4-MALLS-DRi). Food Hydrocoll 74:197–206.  https://doi.org/10.1016/j.foodhyd.2017.08.012 Google Scholar
  16. Luykx DMAM, Peters RJB, van Ruth SM, Bouwmeester H (2008) A review of analytical methods for the identification and characterization of nano delivery systems in food. J Agric Food Chem 56:8231–8247.  https://doi.org/10.1021/jf8013926 Google Scholar
  17. Magarelli M, Passamonti P, Renieri C (2010) Purification, characterization and analysis of sepia melanin from commercial sepia ink (Sepia officinalis). In: Rev. CES Med. Vet. Zootec. http://www.redalyc.org/articulo.oa?id=321428104002. Accessed 6 Oct 2015
  18. Matsuura T, Hino M, Akutagawa S et al (2009) Optical and paramagnetic properties of size-controlled ink particles isolated from Sepia officinalis. Biosci Biotechnol Biochem 73:2790–2792.  https://doi.org/10.1271/bbb.90602 Google Scholar
  19. Meredith P, Sarna T (2006) The physical and chemical properties of eumelanin. Pigment Cell Res 19:572–594.  https://doi.org/10.1111/j.1600-0749.2006.00345.x Google Scholar
  20. Nair JR, Pillai D, Joseph SM, et al (2011) Cephalopod research and bioactive substances. Indian J geo-Mar Sci 40:13–27Google Scholar
  21. Nilsson L (2013) Separation and characterization of food macromolecules using field-flow fractionation: a review. Food Hydrocoll 30:1–11.  https://doi.org/10.1016/j.foodhyd.2012.04.007 Google Scholar
  22. Perna G, Palazzo G, Mallardi A, Capozzi V (2011) Fluorescence properties of natural eumelanin biopolymer. J Lumin 131:1584–1588.  https://doi.org/10.1016/j.jlumin.2011.03.055 Google Scholar
  23. Peters R, ten DG, Bouwmeester H et al (2011) Identification and characterization of organic nanoparticles in food. TrAC Trends Anal Chem 30:100–112.  https://doi.org/10.1016/j.trac.2010.10.004 Google Scholar
  24. Podzimek S (2011) Light Scattering, Size Exclusion Chromatography and Asymmetric Flow Field Flow Fractionation: Powerful Tools for the Characterization of Polymers, Proteins and NanoparticlesGoogle Scholar
  25. Prota G (1988) Progress in the chemistry of melanins and related metabolites. Med Res Rev 8:525–556.  https://doi.org/10.1002/med.2610080405 Google Scholar
  26. Pugh TL, Heller W (1957) Density of polystyrene and polyvinyltoluene latex particles. J Colloid Sci 12:173–180.  https://doi.org/10.1016/0095-8522(57)90004-1 Google Scholar
  27. Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675Google Scholar
  28. Soto-Gómez D, Pérez-Rodríguez P, López-Periago JE, Paradelo M (2016) Sepia ink as a surrogate for colloid transport tests in porous media. J Contam Hydrol 191:88–98.  https://doi.org/10.1016/j.jconhyd.2016.05.005 Google Scholar
  29. Swan GA (1974) Structure, chemistry, and biosynthesis of the Melanins. In: Fortschritte der Chemie Organischer Naturstoffe / Progress in the chemistry of organic natural products. Springer, Vienna, pp 521–582Google Scholar
  30. Zeise L, Addison RB, Chedekel MR (1992) Bio-analytical studies of eumelanins. I. Characterization of melanin the particle. Pigment Cell Res Suppl 2:48–53Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Departamento de Química Analítica y Alimentaria, Área de Química Analítica, Facultad de QuímicaUniversidad de VigoVigoSpain
  2. 2.Ultra Trace Analyses Aquitaine UT2A/ADERAHélioparc Pau-PyrénéesPauFrance
  3. 3.Soil Science and Agricultural Chemistry Group, Department of Plant Biology and Soil Science, Faculty of SciencesUniversity of VigoOurenseSpain
  4. 4.Laboratory of Hydrology and Geochemistry of Strasbourg (LHyGeS)Université de Strasbourg / EOST, CNRSStrasbourgFrance

Personalised recommendations