Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients

ABSTRACT Metabolites of human or microbial origin have the potential to be important biomarkers of the disease state in cystic fibrosis (CF). Clinical sample collection and storage conditions may impact metabolite abundances with clinical relevance. We measured the change in metabolite composition b...

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Autores principales:	Stephen Wandro, Lisa Carmody, Tara Gallagher, John J. LiPuma, Katrine Whiteson
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	biomarkers metabolomics microbial metabolites microbiome reproducibility sample storage Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0f26bdcfea7e42a3926b1d0741fc1352
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spelling	oai:doaj.org-article:0f26bdcfea7e42a3926b1d0741fc13522021-12-02T18:15:43ZMaking It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients10.1128/mSystems.00100-172379-5077https://doaj.org/article/0f26bdcfea7e42a3926b1d0741fc13522017-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00100-17https://doaj.org/toc/2379-5077ABSTRACT Metabolites of human or microbial origin have the potential to be important biomarkers of the disease state in cystic fibrosis (CF). Clinical sample collection and storage conditions may impact metabolite abundances with clinical relevance. We measured the change in metabolite composition based on untargeted gas chromatography-mass spectrometry (GC-MS) when CF sputum samples were stored at 4°C, −20°C, or −80°C with one or two freeze-thaw cycles. Daily measurements were taken for 1 week and then weekly for 4 weeks (4°C) and 8 weeks (−20°C). The metabolites in samples stored at −20°C maintained abundances similar to those found at−80°C over the course of 8 weeks (average change in Bray-Curtis distance, 0.06 ± 0.04) and were also stable after one or two freeze-thaw cycles. However, the metabolite profiles of samples stored at 4°C shifted after 1 day and continued to change over the course of 4 weeks (average change in Bray-Curtis distance, 0.31 ± 0.12). The abundances of several amino acids and other metabolites increased with time of storage at 4°C but remained constant at −20°C. Storage temperature was a significant factor driving the metabolite composition (permutational multivariate analysis of variance: r2 = 0.32 to 0.49, P < 0.001). CF sputum samples stored at −20°C at the time of sampling maintain a relatively stable untargeted GC-MS profile. Samples should be frozen on the day of collection, as more than 1 day at 4°C impacts the global composition of the metabolites in the sample. IMPORTANCE Metabolomics has great potential for uncovering biomarkers of the disease state in CF and many other contexts. However, sample storage timing and temperature may alter the abundance of clinically relevant metabolites. To assess whether existing samples are stable and to direct future study design, we conducted untargeted GC-MS metabolomic analysis of CF sputum samples after one or two freeze-thaw cycles and storage at 4°C and −20°C for 4 to 8 weeks. Overall, storage at −20°C and freeze-thaw cycles had little impact on metabolite profiles; however, storage at 4°C shifted metabolite abundances significantly. GC-MS profiling will aid in our understanding of the CF lung, but care should be taken in studies using sputum samples to ensure that samples are properly stored.Stephen WandroLisa CarmodyTara GallagherJohn J. LiPumaKatrine WhitesonAmerican Society for Microbiologyarticlebiomarkersmetabolomicsmicrobial metabolitesmicrobiomereproducibilitysample storageMicrobiologyQR1-502ENmSystems, Vol 2, Iss 6 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	biomarkers metabolomics microbial metabolites microbiome reproducibility sample storage Microbiology QR1-502
spellingShingle	biomarkers metabolomics microbial metabolites microbiome reproducibility sample storage Microbiology QR1-502 Stephen Wandro Lisa Carmody Tara Gallagher John J. LiPuma Katrine Whiteson Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients
description	ABSTRACT Metabolites of human or microbial origin have the potential to be important biomarkers of the disease state in cystic fibrosis (CF). Clinical sample collection and storage conditions may impact metabolite abundances with clinical relevance. We measured the change in metabolite composition based on untargeted gas chromatography-mass spectrometry (GC-MS) when CF sputum samples were stored at 4°C, −20°C, or −80°C with one or two freeze-thaw cycles. Daily measurements were taken for 1 week and then weekly for 4 weeks (4°C) and 8 weeks (−20°C). The metabolites in samples stored at −20°C maintained abundances similar to those found at−80°C over the course of 8 weeks (average change in Bray-Curtis distance, 0.06 ± 0.04) and were also stable after one or two freeze-thaw cycles. However, the metabolite profiles of samples stored at 4°C shifted after 1 day and continued to change over the course of 4 weeks (average change in Bray-Curtis distance, 0.31 ± 0.12). The abundances of several amino acids and other metabolites increased with time of storage at 4°C but remained constant at −20°C. Storage temperature was a significant factor driving the metabolite composition (permutational multivariate analysis of variance: r2 = 0.32 to 0.49, P < 0.001). CF sputum samples stored at −20°C at the time of sampling maintain a relatively stable untargeted GC-MS profile. Samples should be frozen on the day of collection, as more than 1 day at 4°C impacts the global composition of the metabolites in the sample. IMPORTANCE Metabolomics has great potential for uncovering biomarkers of the disease state in CF and many other contexts. However, sample storage timing and temperature may alter the abundance of clinically relevant metabolites. To assess whether existing samples are stable and to direct future study design, we conducted untargeted GC-MS metabolomic analysis of CF sputum samples after one or two freeze-thaw cycles and storage at 4°C and −20°C for 4 to 8 weeks. Overall, storage at −20°C and freeze-thaw cycles had little impact on metabolite profiles; however, storage at 4°C shifted metabolite abundances significantly. GC-MS profiling will aid in our understanding of the CF lung, but care should be taken in studies using sputum samples to ensure that samples are properly stored.
format	article
author	Stephen Wandro Lisa Carmody Tara Gallagher John J. LiPuma Katrine Whiteson
author_facet	Stephen Wandro Lisa Carmody Tara Gallagher John J. LiPuma Katrine Whiteson
author_sort	Stephen Wandro
title	Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients
title_short	Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients
title_full	Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients
title_fullStr	Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients
title_full_unstemmed	Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients
title_sort	making it last: storage time and temperature have differential impacts on metabolite profiles of airway samples from cystic fibrosis patients
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/0f26bdcfea7e42a3926b1d0741fc1352
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_version_	1718378317007028224

Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients

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