Imperial College London
Alternate

ProfessorTriciaTan

Faculty of Medicine, Department of Metabolism, Digestion and Reproduction

Chair in Metabolic Medicine and Endocrinology
 
 
 
//

Contact

 

+44 (0)20 3313 8038t.tan

 
 
//

Location

 

6N6ECommonwealth BuildingHammersmith Campus

//

Summary

 

Publications

Citation

BibTex format

@article{Cegla:2017:10.1177/0004563216675648,
author = {Cegla, J and Jones, BJ and Howard, J and Kay, R and Creaser, CS and Bloom, SR and Tan, TM},
doi = {10.1177/0004563216675648},
journal = {Annals of Clinical Biochemistry},
pages = {293--296},
title = {The preanalytical stability of glucagon as measured by liquid chromatography tandem mass spectrometry and two commercially available immunoassays},
url = {http://dx.doi.org/10.1177/0004563216675648},
volume = {54},
year = {2017}
}
Download

RIS format (EndNote, RefMan)

TY  - JOUR
AB  - BackgroundOne of the main challenges in the measurement of glucagon is the premise that it is unstable in human plasma. Traditionally, protease inhibitors have been used to prevent its degradation; however, their use is controversial. Here, we investigated the optimal method of sample collection for glucagon, with measurement by liquid chromatography tandem mass spectrometry (LC-MS/MS) and two commercially available immunoassays.MethodsBlood from healthy fasting volunteers (n = 10) was processed under a variety of preanalytical conditions including collection in EDTA vs. lithium heparin tubes and the addition of aprotinin and/or a dipeptidyl-peptidase IV (DPPIV) inhibitor. Additionally, the effect of freeze thaw was assessed. Plasma glucagon concentrations were measured by LC-MS/MS and two commercially available immunoassays (HTRF® sandwich immunoassay, Cisbio and Milliplex MAP Human Metabolic Hormone Panel, Merck Millipore).ResultsA systematic bias of Milliplex > LC-MS/MS > HTRF was noted and plasma glucagon concentrations were significantly different between methods (Milliplex vs. LC-MS/MS P < 0.01; Milliplex vs. HTRF P < 0.0001; LC-MS/MS vs. HTRF P < 0.001). The addition of aprotinin, DPPIV inhibitor or a combination of aprotinin and DPPIV inhibitor had no effect on plasma glucagon concentrations when compared to ‘non-stabilized’ samples or each other. Whether samples were taken in EDTA tubes or lithium heparin tubes made no difference to plasma glucagon concentrations. These findings were consistent for all three methods. Plasma glucagon concentrations were not significantly different after two freeze–thaw cycles (performed on samples in EDTA tubes containing aprotinin and DPPIV inhibitor).ConclusionsThis study demonstrates that glucagon is stable in both EDTA and lithium heparin tubes when stored at −80. Furthermore, the addition of ap
AU  - Cegla,J
AU  - Jones,BJ
AU  - Howard,J
AU  - Kay,R
AU  - Creaser,CS
AU  - Bloom,SR
AU  - Tan,TM
DO  - 10.1177/0004563216675648
EP  - 296
PY  - 2017///
SN  - 1758-1001
SP  - 293
TI  - The preanalytical stability of glucagon as measured by liquid chromatography tandem mass spectrometry and two commercially available immunoassays
T2  - Annals of Clinical Biochemistry
UR  - http://dx.doi.org/10.1177/0004563216675648
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000394655300016&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - http://hdl.handle.net/10044/1/69231
VL  - 54
ER  -
Download