TY - JOUR
T1 - Analysis of volatile human urinary metabolome by solid-phase microextraction in combination with gas chromatography-mass spectrometry for biomarker discovery
T2 - application in a pilot study to discriminate patients with renal cell carcinoma
AU - Monteiro, Márcia
AU - Carvalho, Márcia
AU - Henrique, Rui
AU - Jerónimo, Carmen
AU - Moreira, Nathalie
AU - Bastos, Maria de Lourdes
AU - Pinho, Paula Guedes de
N1 - Funding Information:
This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia) through project Pest-C/EQB/LA0006/2013. Márcia Sá Monteiro acknowledges FCT for her PhD Grant ( SFRH/BD/80518/2011 ). To all financing sources the authors are greatly indebted.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 2014/7
Y1 - 2014/7
N2 - A new and simple analytical approach consisting of headspace-solid phase microextraction (HS-SPME) sampling coupled with gas chromatography-ion trap/mass spectrometry (GC-IT/MS) was developed to study the volatile human urinary metabolome. A central composite design (CCD) was used in the optimisation of extraction conditions. Fibre selection and evaluation of pH influence were performed using an univariate mode and the influence of other parameters, such as the time and temperature of extraction, time of incubation and salt addition, that affect the efficiency of the SPME sampling, was carried out using a CCD. With a sample volume of 2 mL, the optimal conditions in terms of total response values and reproducibility were achieved by performing analyses with a divinylbenzene/polydimethylsiloxane (DVB/PDMS) fibre, in an acidic pH (pH 2) with the addition of 0.59 g of NaCl, allowing the sample to equilibrate for 9 min and extracting at 68 °C for 24 min. The applicability of the optimised method was then tested in a pilot non-target analysis of urine samples obtained from patients with renal cell carcinoma (RCC) and healthy individuals. Chemometric unsupervised analyses performed on the volatile pattern acquired for these samples clearly showed the potential of volatile urinary metabolome to discriminate between RCC and control patients.
AB - A new and simple analytical approach consisting of headspace-solid phase microextraction (HS-SPME) sampling coupled with gas chromatography-ion trap/mass spectrometry (GC-IT/MS) was developed to study the volatile human urinary metabolome. A central composite design (CCD) was used in the optimisation of extraction conditions. Fibre selection and evaluation of pH influence were performed using an univariate mode and the influence of other parameters, such as the time and temperature of extraction, time of incubation and salt addition, that affect the efficiency of the SPME sampling, was carried out using a CCD. With a sample volume of 2 mL, the optimal conditions in terms of total response values and reproducibility were achieved by performing analyses with a divinylbenzene/polydimethylsiloxane (DVB/PDMS) fibre, in an acidic pH (pH 2) with the addition of 0.59 g of NaCl, allowing the sample to equilibrate for 9 min and extracting at 68 °C for 24 min. The applicability of the optimised method was then tested in a pilot non-target analysis of urine samples obtained from patients with renal cell carcinoma (RCC) and healthy individuals. Chemometric unsupervised analyses performed on the volatile pattern acquired for these samples clearly showed the potential of volatile urinary metabolome to discriminate between RCC and control patients.
KW - Central composite design
KW - Gas chromatography-ion trap/mass spectrometry
KW - Headspace solid-phase microextraction
KW - Renal cell carcinoma
KW - Urine
KW - Volatile metabolomic profile
UR - http://www.scopus.com/inward/record.url?scp=84902252993&partnerID=8YFLogxK
U2 - 10.1016/j.ejca.2014.04.011
DO - 10.1016/j.ejca.2014.04.011
M3 - Article
C2 - 24820933
AN - SCOPUS:84902252993
SN - 0959-8049
VL - 50
SP - 1993
EP - 2002
JO - European Journal of Cancer
JF - European Journal of Cancer
IS - 11
ER -