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Expanded metabolite coverage of Saccharomyces cerevisiae extract through improved chloroform/methanol extraction and tert-butyldimethylsilyl derivatization

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We present an improved extraction and derivatization protocol for GCeMS analysis of amino/non-amino
acids in Saccharomyces cerevisiae. Yeast cells were extracted with chloroform: aqueous-methanol (1:1, v/
v) and the resulting non-polar and polar extracts combined and dried for derivatization. Polar and nonpolar
metabolites were derivatized using tert-butyldimethylsilyl (t-BDMS) dissolved in acetonitrile. Using
microwave treatment of the samples, the derivatization process could be completed within 2 h (from
>20 h of the conventional method), providing fully derivatized metabolites that contain multiple derivatizable
organic functional groups. This results in a single derivative from one metabolite, leading to
increased accuracy and precision for identification and quantification of the method. Analysis of combined
fractions allowed the method to expand the coverage of detected metabolites from polar metabolites
i.e. amino acids, organic acids and non-polar metabolites i.e. fatty alcohols and long-chain fatty
acids which are normally non detectable. The recoveries of the extraction method was found at 88 ± 4%,
RSD, N = 3 using anthranilic acid as an internal standard. The method promises to be a very useful tool in
various aspects of biotechnological applications i.e. development of cell factories, metabolomics profiling,
metabolite identification, 13C-labeled flux analysis or semi-quantitative analysis of metabolites in yeast
samples.

Keywords:
Metabolomics
Extraction
Derivatization
Saccharomyces cerevisiae

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Informasi Detail

Judul Seri

Analytical Chemistry Research

No. Panggil

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Penerbit

New York : Elsevier B.V. This is an open access article under the CC BY-NC-ND., 2015

Deskripsi Fisik

Analytical Chemistry Research 6 (2015) 9-16

Bahasa

English

ISBN/ISSN

http://dx.doi.org/10

Klasifikasi

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Tipe Isi

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Tipe Media

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Tipe Pembawa

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Edisi

6 (2015)

Subjek

KIMIA ANALITIK

Info Detail Spesifik

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Pernyataan Tanggungjawab

agus

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