skip to main content

Systematic studies on the determination of Hg-labelled proteins using laser ablation-ICPMS and isotope dilution analysis

Kutscher, Daniel ; Fricker, Mattias ; Hattendorf, Bodo ; Bettmer, Jörg ; Günther, Detlef

Analytical and Bioanalytical Chemistry, 2011, Vol.401(9), pp.2691-2698 [Peer Reviewed Journal]

Full text available

Citations Cited by
  • Title:
    Systematic studies on the determination of Hg-labelled proteins using laser ablation-ICPMS and isotope dilution analysis
  • Author: Kutscher, Daniel ; Fricker, Mattias ; Hattendorf, Bodo ; Bettmer, Jörg ; Günther, Detlef
  • Subjects: ICPMS ; Laser ablation ; Gel electrophoresis ; Protein quantification ; Isotope dilution analysis ; Protein labelling
  • Is Part Of: Analytical and Bioanalytical Chemistry, 2011, Vol.401(9), pp.2691-2698
  • Description: A method was developed for the precise and accurate determination of ovalbumin labelled with p -hydroxy-mercuribenzoic acid (pHMB) using polyacrylamide gel electrophoresis with ns-laser ablation–inductively coupled plasma mass spectrometry. Following systematic optimisation of the ablation process in terms of detection sensitivity, two different quantification strategies were applied: external calibration using standards of the derivatized protein after 13 C + normalization and, as a proof of concept, label-specific isotope dilution analysis (IDA) using pHMB enriched in the isotope 199 Hg. Due to the inhomogeneous distribution of the protein within the gel bands, it could be demonstrated that the IDA approach was superior in terms of precision and accuracy. Furthermore, it permits a reliable quantification, if more complex separation protocols are applied, as typically occurring analyte loss and degradation can be compensated for as soon as complete mixture of spike and sample is achieved. The estimated limit of detection was 160 fmol in the case of ovalbumin. In contrast to earlier studies using metals naturally present in proteins, no loss of mercury was observed during separation under denaturing conditions and other sample preparation steps. Using label-specific IDA, the measured isotope ratios in the gel corresponded to recoveries between 95% and 103%.
  • Language: English
  • Identifier: ISSN: 1618-2642 ; E-ISSN: 1618-2650 ; DOI: 10.1007/s00216-011-5199-5

Searching Remote Databases, Please Wait