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Post-mortem tissue analyses in a patient with succinic semialdehyde dehydrogenase deficiency (SSADHD). I. Metabolomic outcomes

Metabolic Brain Disease volume 35, pages 601–614 (2020)Cite this article

A Correction to this article was published on 18 April 2020

This article has been updated

Abstract

Metabolomic characterization of post-mortem tissues (frontal and parietal cortices, pons, cerebellum, hippocampus, cerebral cortex, liver and kidney) derived from a 37 y.o. male patient with succinic semialdehyde dehydrogenase deficiency (SSADHD) was performed in conjunction with four parallel series of control tissues. Amino acids, acylcarnitines, guanidino- species (guanidinoacetic acid, creatine, creatinine) and GABA-related intermediates were quantified using UPLC and mass spectrometric methods that included isotopically labeled internal standards. Amino acid analyses revealed significant elevation of aspartic acid and depletion of glutamine in patient tissues. Evidence for disruption of short-chain fatty acid metabolism, manifest as altered C4OH, C5, C5:1, C5DC (dicarboxylic) and C12OH carnitines, was observed. Creatine and guanidinoacetic acids were decreased and elevated, respectively. GABA-associated metabolites (total GABA, γ-hydroxybutyric acid, succinic semialdehyde, 4-guanidinobutyrate, 4,5-dihydroxyhexanoic acid and homocarnosine) were significantly increased in patient tissues, including liver and kidney. The data support disruption of fat, creatine and amino acid metabolism as a component of the pathophysiology of SSADHD, and underscore the observation that metabolites measured in patient physiological fluids provide an unreliable reflection of brain metabolism.

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Change history

  • 18 April 2020

    Upon publication, it was noted that five of the on-line supplementary figures had incorrect figure: figure legend associations. These were supplementary Figs.��6, 7, 14, 15, and 23.

Abbreviations

SSADH:

Succinic semialdehyde dehydrogenase

SSADHD:

Succinic semialdehyde dehydrogenase deficiency

PM:

Post-mortem

Cer:

Cerebellum

FrC:

Frontal cortex

Pon:

Pons

PaC:

Parietal cortex

Hip:

Hippocampus

CeC:

Cerebral cortex

Crn:

Creatinine

Cre:

Creatine

Suac:

Succinylacetone

AABA:

Alpha-aminobutyric acid

ASA:

Argininosuccinic acid

Xle:

the sum of isoleucine and leucine (isobaric). Standard 3-letter amino acid abbreviations are employed throughout (e.g., glutamine, gln; serine, ser; etc)

GABA:

4-aminobutyric acid

GHB:

4-hydroxybutyric acid

SSA:

Succinic semialdehyde

4,5-DHHA:

4,5-dihydroxyhexanoic acid

Guac:

Guanidinoacetic acid

4-GBA:

4-guanidinobutyric acid

HC:

Homocarnosine (dipeptide of GABA:L-histidine)

C3DC:

Dicarboxylic acid of 3 carbon chain length (e.g., malonylcarnitine)

C16-OH:

16 carbon length species with one hydroxyl- moiety

C10:2:

10 carbon chain length with 2 unsaturations (di-alkene)

C18:2-OH:

18 carbon length species with 2 unsaturations (di-alkene) and a single hydroxyl- moiety

PheAc:

Phenylacetyl carnitine

Benz:

Benzoyl carnitine

C0:

L-carnitine

FIGLU:

Formiminoglutamic acid

C5DC(C10OH):

e.g., the acylcarnitine peak is a mixture of identical molecular weight fragments, including C5 dicarboxylic acid and C10 monohydroxyl-carnitine (the first shown species predominates in concentration)

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Acknowledgements

We gratefully acknowledge the family of the patient for agreeing to submission of autopsied tissues. We thank Dr. Timothy Fazio and Ms. Christine Fischer, Metabolic Disease Unit, Royal Melbourne Hospital, Victoria, Australia, for procurement, coordination, and shipment of tissues. This study was generously supported by the SSADH Association (www.ssadh.net) and HD 91142 from the National Institute of Child Health, National Institutes of Health (KMG).

Author information

Author notes

  1. Trevor Kirby and Dana C. Walters contributed equally to this work.

Affiliations

  1. Department of Pharmacotherapy, Health Sciences Building Room 210C, College of Pharmacy and Pharmaceutical Sciences, Washington State University, 412 E. Spokane Falls Boulevard, Spokane, WA, 99202-2131, USA

    Trevor Kirby, Dana C. Walters, Madalyn Brown, Jean-Baptiste Roullet & K. Michael Gibson

  2. Department of Clinical Chemistry, Metabolic Unit, Amsterdam University Medical Center (Amsterdam UMC) and VU University Medical Center (VUmc), Amsterdam, the Netherlands

    Erwin Jansen & Gajja S. Salomons

  3. Biochemical Genetics Laboratory, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA

    Coleman Turgeon & Piero Rinaldo

  4. Baylor Scott & White Research Institute, Institute of Metabolic Disease, Dallas, TX, USA

    Erland Arning, Paula Ashcraft & Teodoro Bottiglieri

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  1. Trevor Kirby
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Contributions

All authors contributed to the study conception, design and execution. Data collection and analysis were performed by Madalyn Brown, Coleman Turgeon, Piero Rinaldo, Erland Arning, Paula Ashcraft, Teodoro Bottiglieri, Erwin Jansen and Gajja Salomons. Data reduction, statistical analyses, and the first draft of the manuscript was written by Trevor Kirby and Dana Walters. Final oversight of data analyses, interpretation, and editing of the manuscript were performed by Jean-Baptiste Roullet and K. Michael Gibson. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to K. Michael Gibson.

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Electronic supplementary material

Suppl. Fig. 1
figure 14

Amino acid content in extracts of frontal cortex spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-FrC, post-mortem frontal cortex; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations (PNG 25.7 mb)

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Suppl. Fig. 2
figure 15

Amino acid content in extracts of parietal cortex spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-PaC, post-mortem parietal cortex; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations(PNG 6.44 mb)

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Suppl. Fig. 3
figure 16

Amino acid content in extracts of cerebellum spotted onto filter paper and measured by tandem mass spectrometry.The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-Cer, post-mortem cerebellum; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations(PNG 6.44 mb)

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Suppl. Fig. 4
figure 17

Amino acid content in extracts of pons spotted onto filter paper and measuredby tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-Pon, post-mortem pons; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations(PNG 6.44 mb)

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Suppl. Fig. 5
figure 18

Amino acid content in extracts of hippocampus spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-Hip, post-mortem hippocampus; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations(PNG 6.44 mb)

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Suppl. Fig. 6
figure 19

Amino acid content in extracts of hippocampus spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-Hip, post-mortem hippocampus; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations(PNG 6.44 mb)

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Suppl. Fig. 7
figure 20

Amino acid content in extracts of cerebral cortex spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-Liv, post-mortem liver; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations(PNG 6.44 mb)

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Suppl. Fig. 8
figure 21

Amino acid content in extracts of kidney spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. X-axis values indicate individual amino acid species. Abbreviations: PM-Kid, post-mortem kidney; Xle, sum of leucine and isoleucine (isobaric); SUAC, succinylacetone; AABA, α-aminobutyric acid; ASA, argininosuccinic acid; all other amino acids described using standard three letter abbreviations (PNG 6.44 mb)

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Suppl. Fig. 9
figure 22

Acylcarnitine content in extracts of frontal cortex spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: PM-FrC, post-mortem frontal cortex; CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines) (PNG 6.44 mb)

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Suppl. Fig. 10
figure 23

Acylcarnitine content in extracts of parietal cortex spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: PM-PaC, post-mortem parietal cortex; CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines) (PNG 6.44 mb)

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Suppl. Fig. 11
figure 24

Acylcarnitine content in extracts of cerebellum spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: PM-Cer, post-mortem cerebellum; CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines)(PNG 6.44 mb)

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Suppl. Fig. 12
figure 25

Acylcarnitine content in extracts of pons spotted onto filter paper andmeasured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: PM-Pon, post-mortem pons; CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines)(PNG 6.44 mb)

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Suppl. Fig. 13
figure 26

Acylcarnitine content in extracts of hippocampus spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid; PM-Hip, post-mortem hippocampus. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines)(PNG 6.44 mb)

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Suppl. Fig. 14
figure 27

Acylcarnitine content in extracts of cerebral cortex spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: PM-CeC, post-mortem cerebral cortex; CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines)(PNG 227 kb)

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figure 28

Acylcarnitine content in extracts of liver spotted onto filter paper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: PM-Liv, post-mortem liver; CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines)(PNG 314 kb)

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Acylcarnitine content in extracts of kidney spotted onto filter paper andmeasured by tandem mass spectrometry. The figure depicts the range of control values (n=4;green), with the median of control set to 1.0. Patient values are depicted as a horizontal blackline. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitineester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single ordouble alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC)are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g.,C14-OH. Additional abbreviations employed: PM-Kid, post-mortem kidney; CN, carnitine;salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine;FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of twomolecular ions of identical molecular weight, the data is depicted parenthetically (e.g.C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines).(PNG 6.44 mb)

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Suppl. Fig. 17
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Content of C4OH carnitine in regional brain extracts spotted onto filterpaper and measured by tandem mass spectrometry. The figure depicts the range of control values (n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontal black line. The y-axis depicts “multiples of the reference (control) median”. For the x-axis, carnitine ester chain length is presented as carbon number (e.g., C3-carnitine, C4-carnitine). Single or double alkene species are presented as, for example, C14:1 and C14:2. Dicarboxylic acids (DC) are also presented as chain length, e.g., C12DC. Hydroxylated species are depicted as OH, e.g., C14-OH. Additional abbreviations employed: PM-Liv, post-mortem liver; CN, carnitine; salicyl, salicyl carnitine; PheAc-CN, phenylacetate carnitine; Benz-CN, benzoyl carnitine; FIGLU, formiminoglutamic acid. In those instances in which a marker is composed of two molecular ions of identical molecular weight, the data is depicted parenthetically (e.g. C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines)(PNG 6.44 mb)

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Suppl. Fig. 18
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Content of C5DC carnitine in regional brain extracts spotted onto filter paperand measured by tandem mass spectrometry. The figure depicts the range of control values(n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontalblack line. The y-axis depicts “multiples of the reference (control) median”. Abbreviations:C5DC, C5 dicarboxylic acid carnitine (e.g., glutarylcarnitine); PM, post-mortem; Cer, cerebellum;CeC, cerebral cortex; FrC, frontal cortex; Hip, hippocampus; Pon, pons; PaC, parietal cortex.For C5DC carnitine, the data represents two molecular ions of identical molecular weight (e.g.C5DC(C10OH), representing a mixture of C5 dicarboxylic acid and C10 hydroxyl carnitines);nonetheless, the predominant species here is C5DC carnitine (PNG 6.44 mb)

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Suppl. Fig. 19
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Content of C12OH carnitine in regional brain extracts spotted onto filterpaper and measured by tandem mass spectrometry. The figure depicts the range of controlvalues (n=4; green), with the median of control set to 1.0. Patient values are depicted as ahorizontal black line. The y-axis depicts “multiples of the reference (control) median”.Abbreviations: C12OH, C12 hydroxy carnitine; PM, post-mortem; Cer, cerebellum; CeC, cerebralcortex; FrC, frontal cortex; Hip, hippocampus; Pon, pons; PaC, parietal cortex (PNG 6.44 mb)

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Content of guanidino- species in extracts of liver spotted onto filter paperand measured by tandem mass spectrometry. The figure depicts the range of control values(n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontalblack line. The y-axis depicts “multiples of the reference (control) median”. Abbreviations: Liv,liver; guac, guanidinoacetic acid; cre, creatine; crn, creatinine; PM, post-mortem (PNG 6.44 mb)

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Suppl. Fig. 21
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Content of guanidino- species in extracts of kidney spotted onto filter paperand measured by tandem mass spectrometry. The figure depicts the range of control values(n=4; green), with the median of control set to 1.0. Patient values are depicted as a horizontalblack line. The y-axis depicts “multiples of the reference (control) median”. Abbreviations: Kid,kidney; guac, guanidinoacetic acid; cre, creatine; crn, creatinine; PM, post-mortem (PNG 6.44 mb)

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Quantitation of D- and L-2-hydroxyglutaric acids in pooled brain regions(top panels), as well as liver and kidney (bottom panels) using isotope dilution massspectrometry. Values in top panels represent the sum of all analyses across all brain regions forboth n=4 controls and the patient (regions: cerebellum, frontal and parietal cortices, pons,hippocampus, cerebral cortex, cerebellum). Statistical analyses, two-tailed t test (p=ns for both).Data depicted as mean + SEM. Bottom panels depict the range (mean + SEM) for controls (n=4)controls in both liver (liv) and kidney (kid), as well as the value obtained for the patient Additionalabbreviation: P, patient; C, control. Since only a single measure for the patient was available,statistical evaluation was not undertaken (PNG 6.61 mb)

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Suppl. Fig. 23
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Quantitation of D- and L-2-hydroxyglutaric acids in pooled brain regions(top panels), as well as liver and kidney (bottom panels) using isotope dilution massspectrometry. Values in top panels represent the sum of all analyses across all brain regions forboth n=4 controls and the patient (regions: cerebellum, frontal and parietal cortices, pons,hippocampus, cerebral cortex, cerebellum). Statistical analyses, two-tailed t test (p=ns for both).Data depicted as mean + SEM. Bottom panels depict the range (mean + SEM) for controls (n=4)controls in both liver (liv) and kidney (kid), as well as the value obtained for the patient Additionalabbreviation: P, patient; C, control. Since only a single measure for the patient was available,statistical evaluation was not undertaken (PNG 7.20 mb)

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Kirby, T., Walters, D.C., Brown, M. et al. Post-mortem tissue analyses in a patient with succinic semialdehyde dehydrogenase deficiency (SSADHD). I. Metabolomic outcomes. Metab Brain Dis 35, 601–614 (2020). https://doi.org/10.1007/s11011-020-00550-1

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Keywords

  • GABA
  • γ-Hydroxybutyric acid
  • Metabolomics
  • Amino acids
  • Acylcarnitines