Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
Biomedical copper speciation in relation to Wilson's disease using strong anion exchange chromatography coupled to triple quadrupole inductively coupled plasma mass spectrometry. / Solovyev, Nikolay; Ala, Aftab; Schilsky, Michael; Mills, Craig; Willis, Karl; Harrington, Chris F.
в: Analytica Chimica Acta, Том 1098, 15.02.2020, стр. 27-36.Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
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TY - JOUR
T1 - Biomedical copper speciation in relation to Wilson's disease using strong anion exchange chromatography coupled to triple quadrupole inductively coupled plasma mass spectrometry
AU - Solovyev, Nikolay
AU - Ala, Aftab
AU - Schilsky, Michael
AU - Mills, Craig
AU - Willis, Karl
AU - Harrington, Chris F.
PY - 2020/2/15
Y1 - 2020/2/15
N2 - Biomedical analytical methods often rely on indirect measurements, such as immunoassays, which can lack effective metrological traceability. In the nephelometric determination of ceruloplasmin (Cp), an important protein whose circulating level is altered in Wilson's disease (WD), the anti-Cp antibody used is not specific for the biologically active holoprotein so the assay can overestimate the concentration of Cp due to the presence of the apoprotein. By providing quantitation using elemental standards, the use of strong anion exchange chromatography (SAX) coupled to triple quadrupole inductively coupled plasma mass spectrometry (ICP-MS-MS) can overcome the drawbacks of methods for the measurement of metalloproteins reliant on immunoassays. In the current study, a SAX-ICP-MS-MS method for Cp quantification was designed and tested in samples of blood serum of WD patients and healthy controls. Using standards based on a copper-EDTA complex for calibration, the method provides relatively simple quantification of Cp with the limit of detection of 0.1 μg L−1 (limit of quantification 0.4 μg L−1). The method was also used to investigate the copper species separated by using a 30 kDa cut-off ultrafiltration device. The so-called “exchangeable” copper fraction is considered as an alternative clinical biomarker of WD. Using the designed speciation approach, it was shown that the ultrafiltration method can overestimate the “exchangeable” copper fraction due to a removal of copper from Cp. This was confirmed by comparing the enzymatic activity of the fractions. Thus, the specificity of the “exchangeable” copper test can be ensured only under strict maintenance of ultrafiltration conditions.
AB - Biomedical analytical methods often rely on indirect measurements, such as immunoassays, which can lack effective metrological traceability. In the nephelometric determination of ceruloplasmin (Cp), an important protein whose circulating level is altered in Wilson's disease (WD), the anti-Cp antibody used is not specific for the biologically active holoprotein so the assay can overestimate the concentration of Cp due to the presence of the apoprotein. By providing quantitation using elemental standards, the use of strong anion exchange chromatography (SAX) coupled to triple quadrupole inductively coupled plasma mass spectrometry (ICP-MS-MS) can overcome the drawbacks of methods for the measurement of metalloproteins reliant on immunoassays. In the current study, a SAX-ICP-MS-MS method for Cp quantification was designed and tested in samples of blood serum of WD patients and healthy controls. Using standards based on a copper-EDTA complex for calibration, the method provides relatively simple quantification of Cp with the limit of detection of 0.1 μg L−1 (limit of quantification 0.4 μg L−1). The method was also used to investigate the copper species separated by using a 30 kDa cut-off ultrafiltration device. The so-called “exchangeable” copper fraction is considered as an alternative clinical biomarker of WD. Using the designed speciation approach, it was shown that the ultrafiltration method can overestimate the “exchangeable” copper fraction due to a removal of copper from Cp. This was confirmed by comparing the enzymatic activity of the fractions. Thus, the specificity of the “exchangeable” copper test can be ensured only under strict maintenance of ultrafiltration conditions.
KW - Blood serum
KW - Ceruloplasmin
KW - Copper
KW - Inductively coupled plasma mass spectrometry
KW - Strong anion exchange chromatography
KW - Wilson's disease
KW - Hepatolenticular Degeneration/blood
KW - Copper/blood
KW - Humans
KW - Biomedical Research
KW - Chromatography, Ion Exchange/instrumentation
KW - Mass Spectrometry/instrumentation
KW - FRACTIONATION
KW - METALLOPROTEINS
KW - CERULOPLASMIN
KW - GUIDELINES
KW - SERUM
KW - QUANTITATIVE-ANALYSIS
KW - ICP-MS
KW - ABSOLUTE QUANTIFICATION
KW - PROTEINS
KW - TOOL
UR - http://www.scopus.com/inward/record.url?scp=85075902275&partnerID=8YFLogxK
U2 - 10.1016/j.aca.2019.11.033
DO - 10.1016/j.aca.2019.11.033
M3 - Article
C2 - 31948584
AN - SCOPUS:85075902275
VL - 1098
SP - 27
EP - 36
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
ER -
ID: 53469059