• ADRIANA YURIKO KOGA Postgraduate Program in Pharmaceutical Sciences, Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, Paraná, Brazil.
  • BRUNA CARLETTO Postgraduate Program in Pharmaceutical Sciences, Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, Paraná, Brazil.
  • LEANDRO CAVALCANTE LIPINSKI Department of Medicine, State University of Ponta Grossa, Ponta Grossa, Paraná, Brazil.
  • TRAUDI KLEIN Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, Paraná, Brazil.
  • PAULO VITOR FARAGO Postgraduate Program in Pharmaceutical Sciences, Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, Paraná, Brazil.


Objective: A simple high-performace liquid chromatography method was developed and validated to determine 17-β estradiol in poly (ε-caprolactone) nanocapsules.

Methods: The chromatographic conditions were as follows: C18 GL column with a mobile phase of acetonitrile:water (92:8 v/v) at flow rate of 1.5 mL/min with detection at 280 nm. The evaluated parameters were specificity, linearity, limits of detection and quantification, precision, accuracy, and robustness.

Results: The method was specific and linear (r=0.9982). The limits of detection and quantification were 5.78 μg.mL-1 and 17.54 μg.mL-1, respectively. Suitable accurancy and robustness were obtained. The stability assay showed that pH variation occured after 120 days of storage, and no changes were observed regarding the size and polydispersion parameters. The applicability of the method was evaluated by determining the encapsulation efficiency of the E2 nanocapsules after 120 days of storage. The results showed values >99%.

Conclusion: The results demonstrated the applicability of the developed and validated analytical method.

Keywords: High performance liquid chromatography, estradiol, nanoparticles, nanotechnology


1. Blume-Peytavi U, Kottner J, Sterry W, Hodin MW, Griffths TW, Watson RE, et al. Age-associated skin conditions and diseases: Current perspectives and future options. Gerontologist 2016;56:230-42.
2. Tobin DJ. Introduction to skin aging. J Tissue Viability 2017;26:37-46.
3. Gao WL, Wu LS, Zi JH, Wu B, Li YZ, Song YC, et al. Measurement of serum estrogen and estrogen metabolites in pre- and postmenopausal women with osteoarthritis using high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Braz J Med Biol Res 2015;48:146-53.
4. Fuentes N, Silveyra P. Estrogen receptor signaling mechanisms. Adv Protein Chem Struct Biol 2019;116:135-70.
5. Seleit I, Bakry OA, Repey HS, Ali R. Intrinsic versus extrinsic aging: A histopathological, morphometric and immunohistochemical study of estrogen receptor ? and androgen receptor. Skin Pharmacolol Physiol 2016;29:178-89.
6. Yasar P, Ayaz G, User SD, Gupur G, Muyan M. Molecular mechanism of estrogen-estrogen receptor signaling. Reprod Med Biol 2017;16:4-20.
7. Rzepecki AK, Murase JE, Juran R, Fabi SG, McLellan BN. Estrogen-deficient skin: The role of topical therapy. Int J Womens Dermatol 2019;5:85-90.
8. Cooke PS, Nanjappa MK, Ko C, Prins GS, Hess RA. Estrogens in male physiology. Physiol Rev 2017;97:995-1043.
9. Mondockova V, Adamkovicova M, Lukacova M, Grosskopf B, Babosova R, Galbavy D, et al. The estrogen receptor 1 gene affects bone mineral density and osteoporosis treatment efficiency in Slovak postmenopausal women. Med Genet 2018;19:2-13.
10. Mahmoodzadeh S, Dworatzek E. The role of 17?-estradiol and estrogen receptors in regulation of Ca2 + channels and mitochondrial function in cardiomyocytes. Front Endocrinol 2019;10:1-15.
11. Liao ZH, Huang T, Xiao JW, Gu RC, Ouyang J, Wu G, et al. Estrogen signaling effects on muscle-specific immune responses through controlling the recruitment and function of macrophages and T cells. Skelet Muscle 2019;9:20.
12. Lobo RA. Menopause and aging. In: Yen and Jaffe’s Reproductive Endocrinology. Amsterdam: Elsevier; 2009. p. 325-55.
13. Brufani M, Rizzi N, Meda C, Filocamo L, Ceccaci F, Bartolli VG, et al. Novel locally active estrogens accelerate cutaneous wound healing-part 2. Sci Rep 2017;7:1-36.
14. Ealias AM, Saravanakumar MP. A review on the classification, characterisation, synthesis of nanoparticles and their application. Mater Sci Eng 2017;263:2-14.
15. Radwan F. Nanotechnology and medicine. Mater Sci Nanotechnol 2018;2:10-1.
16. Jeevanandam J, Barhoum A, Chan YS, Dufresne A, Danquah MK. Review on nanoparticles and nanostructured materials: History, sources, toxicity and regulations. Beilstein J Nanotechnol 2018;9:1050-74.
17. Kumari A, Yadav SK, Yadav SC. Biodegradable polymeric nanoparticles based drug delivery systems. Colloids Surf B Biointerfaces 2010;75:1-18.
18. Jarai BM, Kolewe EL, Stillman ZS, Raman N, Fromen CA. Polymeric nanoparticles. In: Nanoparticles for Biomedical Applications. ???: ???; 2020. p. 304-23.
19. Azimi B, Nourpanah P, Rabiee M, Arbab S. Poly (?-caprolactone) fiber: An overview. J Eng Fiber Fabr 2014;9:74-90.
20. Msigala SC, Mabiki FP, Styrishave B, Mdegela RH. Optimized HPLC-UV method for separation, detection and quantification of endocrine disrupting estrogens in low quality water. Int J Chem 2017;9:19-27.
21. Soranganba N, Singh IJ. Simultaneous determination of fish steroidal hormones using RP-HPLC with UV detection by multi-step gradient elution technique. Int J Curr Microbiol App Sci 2018;7:1397-404.
22. ICH. International Conference on the Harmonisation of Technical Requirements for the Registration of Pharmaceuticals for Human Use guidelines, Validation of analytical procedures: Text and Methodology, Q2(R1). Vol. 1. Geneva: ICH; 2005.
23. Agencia Nacional de Vigilância Sanitária. ANVISA, Resolução No. 166, de 24 de Julho de 2017. Diário Oficial [da] República Federativa do Brasil, Brasília, DF; 2017.
24. Rawski RI, Sanecki PT, Kijowska KM, Skital PM, Saletnik DE. Regression analysis in analytical chemistry. Determination and validation of linear and quadratic regression dependencies. S Afr J Chem 2016;69:166-73.
25. Yilmaz B, Kadiouglu Y. Determination of 17 b-estradiol in pharmaceutical preparation by UV spectrophotometry and high performance liquid chromatography methods. Arab J Chem 2017;10:1422-8.
26. Liz MV, Amaral B, Stets S, Nagata N, Peralta-Zamora P. Sensitive estrogens determination in wastewater samples by HPLC and fluorescence detection. J Braz Chem Soc 2017;28:1453-60.
27. Moraes FC, Rossi B, Donatoni MC, Oliveira KT, Pereira EC. Sensitive determination of 17 b-estradiol in river water using a graphene based electrochemical sensor. Anal Chim Acta 2015;881:37-43.
28. Loh XJ. The Effect of pH on the hydrolytic degradation of poly (e-caprolactone)-block-poly (ethylene glycol) copolymers. J Appl Polym Sci 2012;127:1-11.
29. Xu L, Liang H, Yang Y, Yu S. Stability and reactivity: Positive and negative aspects for nanoparticle processing. Chem Rev 2018;118:3209-50.
30. Selvamani V. Stability studies on nanomaterials used in drugs. Characterization and Biology of Nanomaterials for Drug Delivery. Amsterdam: Elsevier; 2019. p. 425-44.
31. Souza PM, Lobo FA, Rosa AH, Fraceto LF. Desenvolvimento de nanocápsulas de poli-e-caprolactona contendo o herbicida atrazina. Quim Nova 2012;35:132-7.
32. Alex AT, Joseph A, Shavi G, Rao JV, Udupa N. Development and evaluation of carboplatin-loaded PCL nanoparticles for intranasal delivery. Drug Deliv 2014;23:2144-53.
33. Melo NF, Campos EV, Franz-Montan M, Paula E, Silva CM, Muruyama CR, et al. Characterization of articaine-loaded poly (?-caprolactone) nanocapsules and solid lipid nanoparticles in hydrogels for topical formulations. J Nanosci Nanotechnol 2018;18:4428-38.
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How to Cite
KOGA, A. Y., B. CARLETTO, L. C. LIPINSKI, T. KLEIN, and P. V. FARAGO. “DEVELOPMENT AND VALIDATION OF AN HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE DETERMINATION OF 17-Β ESTRADIOL IN POLYMERIC NANOPARTICLES”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 14, no. 5, May 2021, pp. 112-6, doi:10.22159/ajpcr.2021.v14i5.41355.
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