PMID- 17418226
OWN - NLM
STAT- MEDLINE
DCOM- 20070725
LR  - 20151119
IS  - 0021-9673 (Print)
IS  - 0021-9673 (Linking)
VI  - 1153
IP  - 1-2
DP  - 2007 Jun 15
TI  - Development of an absolute quantification method targeting growth hormone 
      biomarkers using liquid chromatography coupled to isotope dilution mass 
      spectrometry.
PG  - 300-6
AB  - A method to perform absolute quantification of two biomarkers (IGF-1 and IGFBP-3) 
      of growth hormone abuse has been developed. Isotope dilution is used with 
      synthetically labelled peptides as internal standards. Peptide selection and 
      multiple reaction monitoring design are discussed. A simple sample preparation 
      based on the reduction and alkylation of cysteine residues followed by tryptic 
      digestion provides a sufficient digestion of proteins. Serum samples fortified 
      with increasing amounts of target proteins are analysed by liquid 
      chromatography-tandem mass spectrometry (LC-MS/MS) on a triple quadrupole mass 
      spectrometer. Specificity is ensured by the selection of sequences with no 
      homology in BLAST, as well as retention time deviation check, and ion ratio 
      monitoring. Linearity is studied in terms of calibration curves. These curves for 
      IGFBP-3 and IGF-1 are generated with mean slopes of 0.055 and 0.065, intercepts 
      of 0.107 and -0.011, and with coefficients of correlation of 0.95 and 0.98, 
      respectively. These curves result from the addition of proteins to the serum. 
      Risks of variations related to potential matrix effects are therefore reduced, as 
      well as probable variations related to the digestion steps. The working 
      concentration ranges are 4-10 ng/microl for IGFBP-3 and 2-8 ng/microl for IGF-1. 
      Preliminary data regarding repeatability show that relative standard deviations 
      (RSDs) range between 13 and 32% for IGFBP-3 and between 7 and 29% for IGF-1.
FAU - Kirsch, Stephanie
AU  - Kirsch S
AD  - University of Liege, Chemistry Department, Mass Spectrometry Laboratory, Allee de 
      la Chimie 3, B-6c Sart-Tilman, B-4000 Liege, Belgium. s.kirsch@ulg.ac.be
FAU - Widart, Joelle
AU  - Widart J
FAU - Louette, Joel
AU  - Louette J
FAU - Focant, Jean-Francois
AU  - Focant JF
FAU - De Pauw, Edwin
AU  - De Pauw E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070321
PL  - Netherlands
TA  - J Chromatogr A
JT  - Journal of chromatography. A
JID - 9318488
RN  - 0 (Biomarkers)
RN  - 0 (Insulin-Like Growth Factor Binding Protein 3)
RN  - 67763-96-6 (Insulin-Like Growth Factor I)
RN  - 9002-72-6 (Growth Hormone)
SB  - IM
MH  - Biomarkers/*analysis
MH  - Calibration
MH  - Chromatography, Liquid/*methods
MH  - *Doping in Sports
MH  - Growth Hormone/*blood
MH  - Humans
MH  - Insulin-Like Growth Factor Binding Protein 3/*analysis
MH  - Insulin-Like Growth Factor I/*analysis
MH  - Isotope Labeling/methods
MH  - Tandem Mass Spectrometry/*methods
EDAT- 2007/04/10 09:00
MHDA- 2007/07/26 09:00
CRDT- 2007/04/10 09:00
PHST- 2006/10/03 00:00 [received]
PHST- 2007/03/12 00:00 [revised]
PHST- 2007/03/15 00:00 [accepted]
PHST- 2007/04/10 09:00 [pubmed]
PHST- 2007/07/26 09:00 [medline]
PHST- 2007/04/10 09:00 [entrez]
AID - S0021-9673(07)00539-0 [pii]
AID - 10.1016/j.chroma.2007.03.058 [doi]
PST - ppublish
SO  - J Chromatogr A. 2007 Jun 15;1153(1-2):300-6. doi: 10.1016/j.chroma.2007.03.058. 
      Epub 2007 Mar 21.