Multielemental fractionation in human peripheral blood mononuclear cells by size exclusion liquid chromatography coupled to UV and ICP-MS detection.
An analytical methodology is presented in this work to determine metal-biomolecule complexes size distribution patterns of several elements, among different compounds present in human peripheral blood mononuclear cells (PBMC). A hyphenated technique based on size exclusion chromatography (SEC) coupled online to UV and inductively coupled plasma mass spectrometry (ICP-MS) detection is used. Two different SEC columns with separation ranges between 1,500-1,000,000 relative molecular mass (M(r)) (Nanofilm SEC-250) and 5,000 and 100,000 relative molecular mass (M(r)) (TSK-Gel G2000 SW) are used with 10 mmol/L tris-HCl at pH 7.3 as mobile phase. Retention behavior (retention time and peak-area ratios) remained unchanged for several successive separations. Metal-containing compounds are found to a wide range of M(r). Copper-zinc superoxide dismutase, copper and zinc metallothionein, and copper and zinc transferrin are identified in PBMC samples. A high M(r) (147,000) metal-binding protein containing copper and zinc and a high M(r) (107,000) manganese-binding protein were also found; however, these remained unknown.
Alvarado
Gladys
G
Centro de Química Analítica, Escuela de Química, Facultad de Ciencias, Universidad Central de Venezuela, Apartado de Correos 47102, Caracas 1041-A, Venezuela.
Murillo
Miguel
M
D002138:Calibration; D005591:Chemical Fractionation / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D003300:Copper / Q000737:chemistry; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D008970:Molecular Weight; D013482:Superoxide Dismutase / Q000737:chemistry; D015032:Zinc / Q000737:chemistry
D003300:Copper; D013482:Superoxide Dismutase; D015032:Zinc