Quantitative insulin analysis using liquid chromatography-tandem mass spectrometry in a high-throughput clinical laboratory.
Clin Chem
; 59(9): 1349-56, 2013 Sep.
Article
em En
| MEDLINE
| ID: mdl-23709677
ABSTRACT
BACKGROUND:
Circulating insulin concentrations reflect the amount of endogenous insulin produced by the pancreas and can be monitored to check for insulin resistance. Insulin is commonly measured using immunochemiluminometric assays (ICMA). Unfortunately, differing crossreactivities of the various ICMA antibodies have led to variability in assay results. In contrast, liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based approaches can provide a highly specific alternative to immunoassays.METHODS:
Insulin was extracted from patient serum and reduced to liberate the insulin B chain. Subsequent resolution of the peptide was achieved by LC coupled to triple-quadrupole MS. Selected-reaction monitoring of B-chain transitions was used for quantification. Recombinant human insulin was used as a calibrator and was compared against the National Institute for Biological Standards and Control (NIBSC) reference standard. Bovine insulin and a stable isotopic-labeled ((13)C/(15)N) human insulin B chain were used and compared as internal standards.RESULTS:
The LC-MS/MS assay described herein has been validated according to CLIA guidelines with a limit of detection of 1.8 µIU/mL (10.8 pmol/L) and a limit of quantitation of 3 µIU/mL (18.0 pmol/L). A correlation between the LC-MS/MS assay and a US Food and Drug Administration-approved ICMA was completed for patient samples and the resulting Deming regression revealed good agreement. A reference interval for the assay was established.CONCLUSIONS:
A simple, high-throughput, quantitative LC-MS/MS insulin assay traceable to the NIBSC standard has been successfully developed and validated.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cromatografia Líquida
/
Espectrometria de Massas em Tandem
/
Ensaios de Triagem em Larga Escala
/
Insulina
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Clin Chem
Assunto da revista:
QUIMICA CLINICA
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
Estados Unidos