Direct analysis of [6,6-(2)H2]glucose and [U-(13)C6]glucose dry blood spot enrichments by LC-MS/MS.
A liquid chromatography tandem mass spectrometry (LC-MS/MS) using multiple reaction monitoring (MRM) in a triple-quadrupole scan mode was developed and comprehensively validated for the determination of [6,6-(2)H2]glucose and [U-(13)C6]glucose enrichments from dried blood spots (DBS) without prior derivatization. The method is demonstrated with dried blood spots obtained from rats administered with a primed-constant infusion of [U-(13)C6]glucose and an oral glucose load enriched with [6,6-(2)H2]glucose. The sensitivity is sufficient for analysis of the equivalent to <5μL of blood and the overall method was accurate and precise for the determination of DBS isotopic enrichments.
CNC - Center for Neuroscience and Cell Biology, University of Coimbra, UC Biotech, Parque Tecnológico de Cantanhede, Núcleo 04, Lote 8, 3060-197 Cantanhede, Portugal.
Margarida
M
Coelho
CNC - Center for Neuroscience and Cell Biology, University of Coimbra, UC Biotech, Parque Tecnológico de Cantanhede, Núcleo 04, Lote 8, 3060-197 Cantanhede, Portugal.
Vera M
VM
Mendes
Chronic Diseases Research Center (CEDOC), NOVA Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Rua Câmara Pestana n° 6, 6-A Edifício CEDOC II, 1150-082 Lisboa, Portugal.
Inês S
IS
Lima
CNC - Center for Neuroscience and Cell Biology, University of Coimbra, UC Biotech, Parque Tecnológico de Cantanhede, Núcleo 04, Lote 8, 3060-197 Cantanhede, Portugal; Chronic Diseases Research Center (CEDOC), NOVA Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Rua Câmara Pestana n° 6, 6-A Edifício CEDOC II, 1150-082 Lisboa, Portugal; Institute for Interdisciplinary Research (IIIUC), University of Coimbra, Casa Costa Alemão, Pólo II, Rua Dom Francisco de Lemos, 3030-789 Coimbra, Portugal.
Fátima O
FO
Martins
Chronic Diseases Research Center (CEDOC), NOVA Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Rua Câmara Pestana n° 6, 6-A Edifício CEDOC II, 1150-082 Lisboa, Portugal.
Ana B
AB
Fernandes
Chronic Diseases Research Center (CEDOC), NOVA Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Rua Câmara Pestana n° 6, 6-A Edifício CEDOC II, 1150-082 Lisboa, Portugal; APDP-Portuguese Diabetes Association, Rua Rodrigo da Fonseca, n° 1, 1250-189 Lisboa, Portugal.
M Paula
MP
Macedo
CNC - Center for Neuroscience and Cell Biology, University of Coimbra, UC Biotech, Parque Tecnológico de Cantanhede, Núcleo 04, Lote 8, 3060-197 Cantanhede, Portugal; APDP-Portuguese Diabetes Association, Rua Rodrigo da Fonseca, n° 1, 1250-189 Lisboa, Portugal.
John G
JG
Jones
CNC - Center for Neuroscience and Cell Biology, University of Coimbra, UC Biotech, Parque Tecnológico de Cantanhede, Núcleo 04, Lote 8, 3060-197 Cantanhede, Portugal. Electronic address: bmanadas@gmail.com.
Bruno
B
Manadas
D000818:Animals; D001786:Blood Glucose / Q000032:analysis*; D002247:Carbon Isotopes / Q000008:administration & dosage / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D059788:Dried Blood Spot Testing / Q000379:methods*; D005947:Glucose / Q000008:administration & dosage / Q000493:pharmacokinetics; D057230:Limit of Detection; D016014:Linear Models; D051381:Rats; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D001786:Blood Glucose; D002247:Carbon Isotopes; D005947:Glucose
Animal model; Dried blood spot; Glucose kinetics; Isotope dilution measurement