Microchimica Acta

, 186:647 | Cite as

Biological sample preparation by using restricted-access nanoparticles prepared from bovine serum albumin: application to liquid chromatographic determination of β-blockers

  • Mariana Azevedo Rosa
  • Henrique Dipe De Faria
  • Diailison Teixeira Carvalho
  • Eduardo Costa FigueiredoEmail author
Original Paper


Restricted-access nanoparticles (RANPs) were prepared from bovine serum albumin by coacervation. They have an average sized of 311 nm. They were characterized and used to capture the β-blockers atenolol, metoprolol and propranolol from untreated biological samples. It is shown that both high protein affinity drugs (propranolol) and low protein affinity drugs (atenolol) could be rapidly extracted from plasma. This is revealed by kinetic and isothermal adsorption studies. On the other hand, almost all proteins from the sample were excluded. This demonstrates the efficiency of RANPs as restricted-access material. Sample preparation was carried out by solid phase microextraction using a probe obtained by the fixation of the RANPs at the end of a glass capillary. Atenolol (in concentrations from 100 to 1200 μg L−1), metoprolol (from 80 to 1000 μg L−1) and propranolol (from 15 to 200 μg L−1) were extracted from spiked plasma samples and analyzed by LC MS/MS without using a separation column. Correlation coefficients >0.99, good precision, accuracy, robustness, and lack of memory effects were observed for all of the analytes. The detection limits (at an S/N of 3) are 25.6, 14.6, and 3.8 μg L−1 for atenolol, metoprolol and propranolol, respectively. Ten samples can be simultaneously extracted within ∼15 min. Plasma samples of patients undergoing medical treatment were successfully analyzed with the method.

Graphical abstract

Schematic representation of a bovine serum albumin-based restricted access nanoparticle that exclude proteins from a human plasma sample but capture the small analytes.


Restricted access material Restricted access nanoparticles Solid phase microextraction β-Blockers Liquid chromatography Mass detector Human plasma 



The authors thank Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG, Belo Horizonte, Brazil) [projects CDS-APQ-00638-17 and CDS-PPM-00144-15]; the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brasília, Brazil) [project 427365/2018-0]; and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the financial support.

Compliance with ethical standards

The authors declare that they have no competing interests.

Supplementary material

604_2019_3774_MOESM1_ESM.docx (37 kb)
ESM 1 (DOCX 37.0 kb)


  1. 1.
    de Faria HD, Abrão LC de C, Santos MG et al (2017) New advances in restricted access materials for sample preparation: a review. Anal Chim Acta 959:43–65. CrossRefPubMedGoogle Scholar
  2. 2.
    Kataoka H (2018) Recent advances in online column-switching sample preparation. In: Reference module in chemistry, molecular sciences and chemical engineering. Elsevier, pp 1–30Google Scholar
  3. 3.
    Denadai M, Cass QB (2015) Simultaneous determination of fluoroquinolones in environmental water by liquid chromatography-tandem mass spectrometry with direct injection: a green approach. J Chromatogr A 1418:177–184. CrossRefPubMedGoogle Scholar
  4. 4.
    Gomes RAB, Luccas PO, de Magalhães CS, de Figueiredo EC (2016) Evaluation of the pH influence on protein exclusion by restricted access carbon nanotubes coated with bovine serum albumin. J Mater Sci 51:7407–7414. CrossRefGoogle Scholar
  5. 5.
    Shankar KR, Ameta RK, Singh M (2016) Preparation of BSA nanoparticles using aqueous urea at T = 308.15, 313.15 and 318.15 K as a function of temperature. J Mol Liq 216:808–813. CrossRefGoogle Scholar
  6. 6.
    Tang QS, Chen DZ, Xue WQ et al (2011) Preparation and biodistribution of 188Re-labeled folate conjugated human serum albumin magnetic cisplatin nanoparticles (188Re-folate-CDDP/HSA MNPs) in vivo. Int J Nanomedicine 6:3077–3085. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Elzoghby AO, Samy WM, Elgindy NA (2012) Albumin-based nanoparticles as potential controlled release drug delivery systems. J Control Release 157:168–182. CrossRefPubMedGoogle Scholar
  8. 8.
    Nsubuga H, Basheer C, Haider MB, Bakdash R (2018) Sol-gel based biogenic silica composite as green nanosorbent for chemometric optimization of micro-solid-phase extraction of beta blockers. J Chromatogr A 1554:16–27. CrossRefPubMedGoogle Scholar
  9. 9.
    Galisteo-González F, Molina-Bolívar JA (2014) Systematic study on the preparation of BSA nanoparticles. Colloid Surface B 123:286–292. CrossRefGoogle Scholar
  10. 10.
    Barbosa AF, Barbosa VMP, Bettini J, Luccas PO, Figueiredo EC (2015) Restricted access carbon nanotubes for direct extraction of cadmium from human serum samples followed by atomic absorption spectrometry analysis. Talanta 131:213–220. CrossRefPubMedGoogle Scholar
  11. 11.
    dos Santos RC, Kakazu AK, Santos MG, Belinelli Silva FA, Figueiredo EC (2017) Characterization and application of restricted access carbon nanotubes in online extraction of anticonvulsant drugs from plasma samples followed by liquid chromatography analysis. J Chromatogr B 1054:50–56. CrossRefGoogle Scholar
  12. 12.
    Silva FAB, Chagas-Silva FA, Florenzano FH, Pissetti FL (2016) Poly(dimethylsiloxane) and poly[vinyltrimethoxysilane-co-2-(dimethylamino) ethyl methacrylate] based cross-linked oroganic-inorganic hybrid adsorbent for copper(II) removal from aqueous solutions. J Braz Chem Soc 27:2181–2191. CrossRefGoogle Scholar
  13. 13.
    Foo KY, Hameed BH (2010) Insights into the modeling of adsorption isotherm systems. Chem Eng J 156:2–10. CrossRefGoogle Scholar
  14. 14.
    Boscari CN, Mazzuia GR, Wisniewski C, Borges KB, Figueiredo EC (2017) Molecularly imprinted probe for solid-phase extraction of hippuric and 4-methylhippuric acids directly from human urine samples followed by MEKC analysis. Electrophoresis 38:1083–1090. CrossRefPubMedGoogle Scholar
  15. 15.
    FDA - U.S. Department of Health and Human Services (2013) Guidance for industry: bioanalytical method validation. 34.
  16. 16.
    Rahimnejad M, Najafpour G, Bakeri G (2012) Investigation and modeling effective parameters influencing the size of BSA protein nanoparticles as colloidal carrier. Colloid Surface A 412:96–100. CrossRefGoogle Scholar
  17. 17.
    Menezes ML, Fèlix G (1998) On line extraction and separation of bendiocarb, methomyl, methylparathion, and pentachlorophenol pesticides from raw milk. J Liq Chromatogr Relat Technol 21:2863–2871. CrossRefGoogle Scholar
  18. 18.
    Bronze-Uhle ES, Costa BC, Ximenes VF, Lisboa-Filho PN (2017) Synthetic nanoparticles of bovine serum albumin with entrapped salicylic acid. Nanotechnol Sci Appl 10:11–21. CrossRefPubMedGoogle Scholar
  19. 19.
    Kumar KV, Sivanesan S (2006) Pseudo second order kinetics and pseudo isotherms for malachite green onto activated carbon: comparison of linear and non-linear regression methods. J Hazard Mater 136:721–726. CrossRefPubMedGoogle Scholar
  20. 20.
    Liu Y, Liu Y-J (2008) Biosorption isotherms, kinetics and thermodynamics. Sep Purif Technol 61:229–242. CrossRefGoogle Scholar
  21. 21.
    Kufleitner J, Wagner S, Worek F, von Briesen H, Kreuter J (2010) Adsorption of obidoxime onto human serum albumin nanoparticles: drug loading, particle size and drug release. J Microencapsul 27:506–513. CrossRefPubMedGoogle Scholar
  22. 22.
    Figueiredo EC, Sparrapan R, Sanvido GB, Santos MG, Zezzi Arruda MA, Eberlin MN (2011) Quantitation of drugs via molecularly imprinted polymer solid phase extraction and electrospray ionization mass spectrometry: benzodiazepines in human plasma. Analyst 136:3753–3757. CrossRefPubMedGoogle Scholar
  23. 23.
    Benfield P, Clissold SP, Brogden RN (1986) Metoprolol: an updated review of its Pharmacodynamic and pharmacokinetic properties, and therapeutic efficacy, in hypertension, Ischaemic heart disease and related cardiovascular disorders. Drugs 31:376–429. CrossRefPubMedGoogle Scholar
  24. 24.
    Leonetti G, Terzoli L, Bianchini C, Sala C, Zanchetti A (1980) Time-course of the anti-hypertensive action of atenolol: comparison of response to first dose and to maintained oral administration. Eur J Clin Pharmacol 18:365–374. CrossRefPubMedGoogle Scholar
  25. 25.
    Bengtsson C, Johnson G, Regdh CG (1975) Plasma levels and effects of metoprolol on blood pressure and heart rate in hypertensive patients after an acute dose and between two doses during long-term treatment. Clin Pharmacol Ther 17:400–408. CrossRefPubMedGoogle Scholar
  26. 26.
    Mansur AP, Avakian SD, Paula RS, Donzella H, Santos SRCJ, Ramires JAF (1998) Pharmacokinetics and pharmacodynamics of propranolol in hypertensive patients after sublingual administration: systemic availability. Brazilian J Med Biol Res 31:691–696. CrossRefGoogle Scholar
  27. 27.
    Fan W, He M, You L, Zhu X, Chen B, Hu B (2016) Water-compatible graphene oxide/molecularly imprinted polymer coated stir bar sorptive extraction of propranolol from urine samples followed by high performance liquid chromatography-ultraviolet detection. J Chromatogr A 1443:1–9. CrossRefPubMedGoogle Scholar
  28. 28.
    Gorbani Y, Yılmaz H, Basan H (2017) Spectrofluorimetric determination of atenolol from human urine using high-affinity molecularly imprinted solid-phase extraction sorbent. Luminescence 32:1391–1397. CrossRefPubMedGoogle Scholar
  29. 29.
    Ensafi AA, Kazemifard N, Rezaei B (2017) Development of a nano plastic antibody for determination of propranolol using CdTe quantum dots. Sensor Actuat B-Chem 252:846–853. CrossRefGoogle Scholar
  30. 30.
    Kim HM, Park JH, Long NP, Kim DD, Kwon SW (2019) Simultaneous determination of cardiovascular drugs in dried blood spot by liquid chromatography-tandem mass spectrometry. J Food Drug Anal:1–9 (in press).
  31. 31.
    Elgawish MS, Mostafa SM, Elshanawane AA (2011) Simple and rapid HPLC method for simultaneous determination of atenolol and chlorthalidone in spiked human plasma. Saudi Pharm J 19:43–49. CrossRefPubMedGoogle Scholar
  32. 32.
    Renkecz T, Ceolin G, Horváth V (2011) Selective solid phase extraction of propranolol on multiwell membrane filter plates modified with molecularly imprinted polymer. Analyst 136:2175–2182. CrossRefPubMedGoogle Scholar
  33. 33.
    Jamshidi S, Rofouei MK, Thorsen G (2019) Using magnetic core-shell nanoparticles coated with an ionic liquid dispersion assisted by effervescence powder for the micro-solid-phase extraction of four beta blockers from human plasma by ultra high performance liquid chromatography with mass spectrom. J Sep Sci 42:698–705. CrossRefPubMedGoogle Scholar
  34. 34.
    Mabrouk MM, Hammad SF, El-Malla SF, Elshenawy EA (2019) Green micellar HPLC-fluorescence method for simultaneous determination of metoprolol and amlodipine in their combined dosage form: application on metoprolol in spiked human plasma. Microchem J 147:635–642. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratory of Toxicant and Drug AnalysesFederal University of Alfenas – Unifal-MGAlfenasBrazil

Personalised recommendations