Metalloprotein separation and analysis by directly coupled size exclusion high-performance liquid chromatography inductively coupled plasma mass spectroscopy

Journal Article · Mon Jan 01 04:00:00 EST 1990 · Analytical Biochemistry; (United States)

DOI:https://doi.org/10.1016/0003-2697(90)90066-I· OSTI ID:6016387

Mason, A Z; Storms, S D; Jenkins, K D ^[1]

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1. California State Univ., Long Beach (United States)

The feasibility of using directly coupled size exclusion high-performance liquid chromatography inductively coupled plasma mass spectroscopy (HPLC/ICP-MS) for the separation and subsequent elemental analysis of metalloproteins in biological samples has been studied. Data, on up to eight elements, was acquired simultaneously and the reconstructed elemental profiles from the chromatographed samples were quantified by flow injection analysis. Absolute and relative detection limits, reproducibility, operational dynamic range, and linearity of response were initially evaluated by analyzing standards of metallothionein protein of known elemental composition for Cd, Zn, and Cu. There was evidence of displacement of Zn from the protein during chromatography and the substitution of Cu sequestered from the mobile phase. Cd associated with the protein was fully recovered during chromatography. The versatility of the technique has been demonstrated by the quantitative multielement analysis of cytosolic metal-binding proteins separated from the polychaete worm Neanthes arenaceodentata. Fidelity of analysis has been demonstrated by two independent procedures: first, by comparing the elemental profiles obtained by directly aspirating the HPLC eluant into the ICP-MS to those obtained by collecting fractions and quantifying the metal content of the proteins in the conventional analytical mode; second, by comparing the stable isotopic profiles for {sup 114}Cd obtained by simultaneous ICP-MS analysis with radiometric profiles of {sup 109}Cd obtained by counting radioactivity associated with collected fractions. The virtues and limitations of ICP-MS as a multielement HPLC detector for the analysis of metalloproteins are described and compared to more established techniques using other element specific or multielement detectors. 60 refs., 7 figs., 9 tabs.

• DOE Contract Number: FG03-86ER60495
• OSTI ID: 6016387
• Journal Information: Analytical Biochemistry; (United States), Journal Name: Analytical Biochemistry; (United States) Vol. 186; ISSN ANBCA; ISSN 0003-2697
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
400102* -- Chemical & Spectral Procedures
ABSORPTION SPECTROSCOPY
ABUNDANCE
BETA DECAY RADIOISOTOPES
CADMIUM
CADMIUM 109
CADMIUM 114
CADMIUM ISOTOPES
CHEMICAL ANALYSIS
CHROMATOGRAPHY
COMPARATIVE EVALUATIONS
COPPER
DATA
ELECTRON CAPTURE RADIOISOTOPES
ELEMENT ABUNDANCE
ELEMENTS
EVALUATION
EVEN-EVEN NUCLEI
EVEN-ODD NUCLEI
EXPERIMENTAL DATA
INFORMATION
INTERMEDIATE MASS NUCLEI
ISOTOPES
LIQUID COLUMN CHROMATOGRAPHY
MASS SPECTROSCOPY
METALLOPROTEINS
METALLOTHIONEIN
METALS
NUCLEI
NUMERICAL DATA
ORGANIC COMPOUNDS
PLASMA
PROTEINS
QUANTITATIVE CHEMICAL ANALYSIS
RADIOISOTOPES
RADIOMETRIC ANALYSIS
SAMPLE PREPARATION
SEPARATION PROCESSES
SPECTROSCOPY
STABLE ISOTOPES
TRANSITION ELEMENTS
YEARS LIVING RADIOISOTOPES
ZINC