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.
ᅟ
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
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
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
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
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
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 253–323
Derby CD (2014) Cephalopod ink: production, chemistry, functions and applications. Mar Drugs 12:2700–2730. https://doi.org/10.3390/md12052700
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
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
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
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
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
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
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 213:2353–2361
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
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
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
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
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
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
Nair JR, Pillai D, Joseph SM, et al (2011) Cephalopod research and bioactive substances. Indian J geo-Mar Sci 40:13–27
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
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
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
Podzimek S (2011) Light Scattering, Size Exclusion Chromatography and Asymmetric Flow Field Flow Fractionation: Powerful Tools for the Characterization of Polymers, Proteins and Nanoparticles
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
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
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675
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
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 521–582
Zeise L, Addison RB, Chedekel MR (1992) Bio-analytical studies of eumelanins. I. Characterization of melanin the particle. Pigment Cell Res Suppl 2:48–53
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
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).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
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.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Highlights
- Eumelanin nanoparticles were isolated from commercial sepia ink.
- Sepia ink nanoparticles can be used as a model for organic nanoparticles studies.
- SEM showed quasi-spherical particles of 100–140 nm diameter.
- DLS indicated an apparent hydrodynamic diameter of 180–260 nm.
- AF4-MALLS provides a hydrodynamic diameter of ≈ 200 nm.
Electronic Supplementary Material
ESM 1
(DOCX 1540 kb)
Rights and permissions
About this article
Cite this article
de la Calle, I., Soto-Gómez, D., Pérez-Rodríguez, P. et al. Particle Size Characterization of Sepia Ink Eumelanin Biopolymers by SEM, DLS, and AF4-MALLS: a Comparative Study. Food Anal. Methods 12, 1140–1151 (2019). https://doi.org/10.1007/s12161-019-01448-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12161-019-01448-0