TY - JOUR
T1 - High sensitivity LC-MS/MS method for direct quantification of human parathyroid 1-34 (teriparatide) in human plasma
AU - Chambers, Erin E.
AU - Lame, Mary E.
AU - Bardsley, Jon
AU - Hannam, Sally
AU - Legido-Quigley, Cristina
AU - Smith, Norman
AU - Fountain, Kenneth J.
AU - Collins, Eileen
AU - Thomas, Elizabeth
N1 - © 2013 Elsevier B.V.
PY - 2013/11/1
Y1 - 2013/11/1
N2 - Teriparatide, the 1-34 fragment of human parathyroid hormone, is used to treat osteoporosis patients with a high risk of fracture by stimulating new bone formation. Routinely teriparatide is quantified using radioimmunoassay however the LC-MS/MS described here has the potential to achieve greater accuracy and precision, higher specificity, and is readily implemented in routine bioanalytical laboratories. Hence a complete method combining effective sample prep with appropriate LC separation and selected reaction monitoring (SRM) MS detection was developed to selectively separate teriparatide from closely related endogenous peptides and to reduce interferences. Samples were concentrated without evaporation, minimizing the risk of adsorptive losses. Chromatography was performed on a sub 2. μm particle charged surface hybrid column, which provided significantly higher peak capacity than a traditional C18 column when formic acid was used as the mobile phase modifier. Total LC cycle time was 6. min. An LOD of 15. pg/mL (3.6. fmol/mL) from 200. μL of human plasma was readily achieved and standard curves were accurate and precise from 15. pg/mL to 500. pg/mL. Mean QC accuracies ranged from 90% to 106%. Mean QC precision was better than 7%. The CV of matrix factors across 6 sources of human plasma was 5%. The assay presented here is the first LC-MS method which reaches clinically relevant detection limits for teriparatide.
AB - Teriparatide, the 1-34 fragment of human parathyroid hormone, is used to treat osteoporosis patients with a high risk of fracture by stimulating new bone formation. Routinely teriparatide is quantified using radioimmunoassay however the LC-MS/MS described here has the potential to achieve greater accuracy and precision, higher specificity, and is readily implemented in routine bioanalytical laboratories. Hence a complete method combining effective sample prep with appropriate LC separation and selected reaction monitoring (SRM) MS detection was developed to selectively separate teriparatide from closely related endogenous peptides and to reduce interferences. Samples were concentrated without evaporation, minimizing the risk of adsorptive losses. Chromatography was performed on a sub 2. μm particle charged surface hybrid column, which provided significantly higher peak capacity than a traditional C18 column when formic acid was used as the mobile phase modifier. Total LC cycle time was 6. min. An LOD of 15. pg/mL (3.6. fmol/mL) from 200. μL of human plasma was readily achieved and standard curves were accurate and precise from 15. pg/mL to 500. pg/mL. Mean QC accuracies ranged from 90% to 106%. Mean QC precision was better than 7%. The CV of matrix factors across 6 sources of human plasma was 5%. The assay presented here is the first LC-MS method which reaches clinically relevant detection limits for teriparatide.
KW - Bioanalysis
KW - Human growth hormone
KW - LC-MS/MS peptide quantification
KW - RhPTH (1-34)
KW - Solid phase extraction (SPE)
KW - Teriparatide
UR - http://www.scopus.com/inward/record.url?scp=84884924592&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2013.08.027
DO - 10.1016/j.jchromb.2013.08.027
M3 - Article
C2 - 24076523
AN - SCOPUS:84884924592
SN - 1570-0232
VL - 938
SP - 96
EP - 104
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical & Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical & Life Sciences
ER -