Development of a quantitative method for active epidermal growth factor extracted from dissolving microneedle by solid phase extraction and liquid chromatography electrospray ionization mass spectrometry

Suyong Kim, Minjoo Park, Huisuk Yang, Manita Dangol, Shayan F. Lahiji, Inyoung Huh, Miroo Kim, Jaeick Lee, Junghyun Son, Hyungil Jung

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Dissolving microneedle (DMN), a transdermal drug delivery in which biological drugs are encapsulated in biodegradable and biocompatible polymers, was fabricated using epidermal growth factor (EGF) as a model drug and hyaluronic acid (HA) as a backbone polymeric matrix. After mixing calibration and DMN samples with insulin, an internal standard, solid phase extraction (SPE) was performed to separate EGF and insulin from HA, and then liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) was conducted for microgram-scale quantitation. The method showed good linearity (R2 = 0.997) within a specified range (1–4 μg). Additionally, the decrease in EGF levels during DMN fabrication was compared using the SPE/LC-ESI-MS and enzyme-linked immunosorbent assay (ELISA), a traditional analytical method. The ELISA method detected an EGF loss of only 3.88 ± 4.67%, whereas SPE/LC-ESI-MS detected a loss of 16.75 ± 4.39%. Qualitative analysis by circular dichroism showed wavelength shift and splitting after DMN fabrication indicating that EGF was denatured during DMN fabrication and cell viability test showed SPE/LC-ESI-MS is more accurate and reliable for detecting the amount of active EGF loaded into the DMN than ELISA.

Original languageEnglish
Pages (from-to)297-302
Number of pages6
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume131
DOIs
Publication statusPublished - 2016 Nov 30

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All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

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