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.
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 (1-4μg). 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.
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea; Juvic Biotech. Inc., Building 102, Yonsei Engineering Research Park, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Suyong
S
Kim
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Minjoo
M
Park
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Huisuk
H
Yang
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Manita
M
Dangol
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Shayan
S
F Lahiji
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Inyoung
I
Huh
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Miroo
M
Kim
Doping Control Center, Korea Institute of Science and Technology (KIST), Hwarang-ro 14-gil 5, Seongbuk-gu, Seoul, 02792, Republic of Korea.
Jaeick
J
Lee
Doping Control Center, Korea Institute of Science and Technology (KIST), Hwarang-ro 14-gil 5, Seongbuk-gu, Seoul, 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of Korea. Electronic address: son.junghyun@kist.re.kr.
Junghyun
J
Son
Department of Biotechnology, Building 123, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea; Juvic Biotech. Inc., Building 102, Yonsei Engineering Research Park, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea. Electronic address: hijung@yonsei.ac.kr.
Hyungil
H
Jung
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002853:Chromatography, Liquid / Q000379:methods*; D002942:Circular Dichroism; D016503:Drug Delivery Systems; D004815:Epidermal Growth Factor / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology; D006820:Hyaluronic Acid / Q000737:chemistry; D051379:Mice; D009339:Needles; D052616:Solid Phase Extraction / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D004815:Epidermal Growth Factor; D006820:Hyaluronic Acid
Dissolving microneedle; Enzyme-linked immunosorbent assay; Epidermal growth factor; Hyaluronic acid; Liquid chromatography electrospray ionization mass spectrometry; Solid phase extraction