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Key data of our analytical platforms are described below and include PubChem numbers, reported biomarker concentrations, LODs, CVs, ICCs, sample matrix and method description for most of our biomarkers.*

Analytical platforms

Short-chain fatty acids.

Sample volume required: 50 µL

All concentrations are reported in µmol/L.

AcetateAceShort-chain fatty acid175-1765-3005.03-53-7s, p
ButyrateButShort-chain fatty acid2641-300.23-63-6s, p
FormateFrmShort-chain fatty acid2848-1007.03-63-6s, p, csf
IsobutyrateiButShort-chain fatty acid65900.7-60.053-63-8s, p, csf
IsovalerateiVlrShort-chain fatty acid104300.3-30.023-63-8s, p
PropionateProShort-chain fatty acid10421-130.062-63-6s, p
ValerateVlrShort-chain fatty acid79910.2-10.053-64-8s, p
α-MethylbutyrateaMBShort-chain fatty acid83140.3-30.053-74-7s, p
β-Hydroxy β-methylbutyrateHMBLeu metabolite693620.1-10, 2-6 (csf)s, p, csf

Amino- and carboxylic acids. TCA metabolites and intermediates.

Sample volume required: 50 µL

All concentrations are reported in µmol/L.

Methylmalonic acidMMACarboxylic acid487<, p
Total homocysteinetHcySulfur amino acid915525-, p
Total cysteinetCysSulfur amino acid5862150-35050120.62s, p
MethionineMetEssential amino acid613718-5012.24.10.33s, p
SerineSerPolar amino acid595195-20051.31.70.71s, p
GlycineGlyHydrophobic amino acid750150-35031.31.70.81s, p
CystathionineCystaThioether4392580.07-0.550., p
Sarcosine*SarcMethylated amino acid10880.7-2.30.0351.31.60.68s, p
HistidineHisEssential amino acid627440-1501230.44as, p
TryptophanTrpEssential aromatic amino acid630540-900., p
KynurenineKynTrp metabolite1611661.0-, p
OrnithineOrnCharged amino acid626230-9012.22.70.55as, p
Aspartic acidAspCharged amino acid59600-260.51.84s, p
Glutamic acidGluCharged amino acid3303220-1400.51.72.7s, p
LysineLysEssential amino acid5962120-2901., p
AlanineAlaHydrophobic amino acid5950230-51011.21.40.43as, p
PhenylalaninePheEssential aromatic amino acid614026-850.51.530.44as, p
IsoleucineIleEssential BCAA**630640-1400., p
LeucineLeuEssential BCAA**610670-1700.53.440.52as, p
ProlineProHydrophobic amino acid145742110-3600., p
ValineValEssential BCAA**6287150-3500., p
AsparagineAsnPolar amino acid626720-1300.6330.40as, p
GlutamineGlnPolar amino acid5961390-700102.12.70.42as, p
PhenylacetylglutaminePAGlnn-acyl-alpha amino acid922582-10015-66-7s, p
ThreonineThrEssential amino acid628870-2401., p
TyrosineTyrAromatic amino acid605740-1100., p
α-Hydroxyglutaric acidaHGα-Hydroxy acid430.15-, p
3-Hydroxyisobutyrate3HIBVal metabolite44087315-2512.42.9s, p
2-HydroxybutyrateaHBα-Hydroxy acid1126610-6011.72.3s, p
3-HydroxybutyratebHBKetone body44120-70011.93.4s, p
AcetoacetateAcAcKetone body9610-200555s, p
CarboxymethyllysineCMLan AGE***1238000.05-, p
CarboxyethyllysineCELan AGE***234007790.05-, p
2-Aminoadipic acid2AAALys metabolite4690.3-, p
β-AminoisobutyrateBAIBAVal metabolite649560.3-, p
β-AlaninebAlaβ-Amino acid2390.5-100.334.3s, p
CitrateCitTricarboxylic acid31190-200433.1s, p
IsocitrateiCitTricarboxylic acid11982-220.32.83s, p, csf
α-KetoglutarateaKGKeto acid512-400.53.73.7s, p, csf
FumarateFumDicarboxylic acid4449720.5-, p, csf
MalateMalβ-Hydroxy acid5252-, p, csf
PyruvatePyrα-Keto acid106080-160566.5s, p, csf
LactateLacα-Hydroxy acid612500-1600104.64.8s, p, csf

*Sarcosine cannot be measured in EDTA-tubes from some suppliers because of the presence of sarcosine in these tubes; **BCAA, Branched chain amino acid; ***AGE, Advanced glycation end product.
aXiaofei Yin, Orla Prendiville, Aoife E. McNamara, and Lorraine Brennan. Targeted Metabolomic Approach to Assess the Reproducibility of
Plasma Metabolites over a Four Month Period in a Free-Living Population. J. Proteome Res. 2022, 21, 683−690.

Choline derivatives. Charged, methylated or sulfur amino acids.

Sample volume required: 35 µL

All concentrations are reported in µmol/L.

CholineCholQuarternary ammonium3055–, p, u
Trimethylamine N-oxideTMAOAmine oxide11450.5–600.144.1<0.38s, p, u
BetaineBetMethylated amino acid24715–600., p, u
DimethylglycineDMGMethylated amino acid6731.5–, p, u
Total homocysteinetHcyAmino acid91552<150.1120.72s, p, u
CreatineCrnα-Amino acid58610–10013.94s, p, u
CreatinineCreatα-Amino acid58850–1000., p, u
MethionineMetEssential amino acid613718–5014.14.50.33s, p
Methionine sulfoxideMetSoPolar amino acid1589800.5–, p
Total cysteinetCysSulfur amino acid5862150–4000.1260.56s, p
HistidineHisEssential amino acid627440–2000.253.53.8s, p
3-Methylhistidine3-MHAmino acid921052.7–, p, u
1-Methylhistidine1-MHAmino acid649690.5–, p, u
CitrullineCtnAmino acid975010–6047-86-9s, p
ArginineArgCharged amino acid632210–1000.252.630.53s, p
Asymmetric dimethylarginineADMAGuanidinated amino acid1238310.4–, p
Symmetric dimethylarginineSDMAGuanidinated amino acid1691480.3–, p
HomoargininehArgGuanidinated amino acid90851–60.15.860.65s, p
TrimethyllysineTMLMethylated amino acid4401210.3–, p
CarnitineC0Carnitine28820–800.50.69as, p
Carnitine, totalbtC0Carnitines30–900.5s, p

aFloegel A, Drogan D, Wang-Sattler R, Prehn C, Illig T, et al. (2011) Reliability of Serum Metabolite Concentrations over a 4-Month Period Using a
Targeted Metabolomic Approach. PLoS ONE 6(6): e21103. doi:10.1371/journal.pone.0021103
bMeasured in a separate assay involving alkaline hydrolysis of acylcarnitines.

Kynurenine pathway and B vitamins (B1, B2, B3, B6), indoles, nicotin, coffee markers, and inflammation markers.

Sample volume required: 60 µL

3-Hydroxyanthranilic acidHAATrp metabolite8610–80/0–7 nmol/L24.660.54s, p, csf
3-HydroxykynurenineHKTrp metabolite1181125–80/0.1–25 nmol/L23.24.50.62s, p, csf
3-Indoxyl sulfate3ISIndole, Trp metabolite102580.9–30 µmol/L0.2s, p, csf
4-Pyridoxic acidPAB6 vitamer672310–200/0–15 nmol/L0., p, csf
Anthranilic acidAATrp metabolite2277–30/3–14 nmol/L27.49.50.64s, p, csf
CotinineCotNicotine metabolite854019Non-smokers < ~5 nmol/L; passive smokers 5 – 85 nmol/L; smokers > 85nmol/L; heavy smokers (>25 cigarettes/day) > 1700 nmol/L134.20.92s, p,csf
CystathionineCystaThioether43925870–550 nmol/L0., p, csf
Flavin mononucleotideFMNB2 vitamer6439763–30/1–2 nmol/L0., p, csf
Imidazole propionateImPHistidine metabolite706303–500 nmol/L2s, p
Indole-3-acetamideIAMIndole, Trp metabolite3970–10 nmol/L1s, p
Indole-3-acetateIAAIndole, Trp metabolite8020.3–23 µmol/L0.05s, p
Indole-3-aldehydeIAldIndole, Trp metabolite1025610–200 nmol/L4s, p
Indole-3-lactateILAIndole, Trp metabolite929040.08–5 µmol/L0.025s, p
Indole-3-propionateIPAIndole, Trp metabolite37440.5–12 µmol/L0.04s, p
Kynurenic acidKATrp metabolite384520–100/0.5–15 nmol/L0., p, csf
KynurenineKynTrp metabolite1611661.0–3.3/0.02–0.12 µmol/L0.0073.65.10.68s, p, csf
N1-methylnicotinamidemNAMB3 vitamer45720–250/5–100 nmol/L545.5s, p, csf
NeopterinNeoptPteridine44555–50/5–50 nmol/L0., p, csf
NicotinamideNAMB3 vitamer936100–600/5–200 nmol/L204.26.1s, p, csf
Nicotinic acidNAB3 vitamer938<20 nmol/L1045.4s, p, csf
Picolinic acidPicTrp metabolite101820–100/5–100 nmol/L86-75-8s, p, csf
PyridoxalPLB6 vitamer10505–150/10–50 nmol/L0., p, csf
Pyridoxal 5-phosphatePLPB6 vitamer105115–150/7–70 nmol//L0., p, csf
PyridoxinePNB6 vitamer1054Exogenous metabolite, reflecting B vitamin supplement use0.54.55.2s, p
Quinaldic acidQldTrp (KA) metabolite71240.2–100/0.1–10 nmol/L0.56-76-8s, p
Quinolinic acidQATrp metabolite1066150–700/10–150 nmol/L1.67-87-10s, p, csf
RiboflavinRiboB2 vitamer4935705–100/5–25 nmol/L0.267.10.79s, p, csf
ThiamineThiB1 vitamer11302–50/5–40 nmol/L0.53.54.9s, p, csf
Thiamine monophosphateTMPB1 vitamer11312–15/25–75 nmol/L0.57.111.6s, p, csf
Trans-3-hydroxycotinineOHCotNicotine metabolite107963Concentrations relative to cotinine, with OHCot/cotininine ratio ranging from 0.02-0.9023.64.50.85s, p, csf
TrigonellineTrigAlkaloid55700.4–25 µmol/L0., p, csf
Trimethylamine N-oxideTMAOAmine oxide11450.5–60/0.2–12 µmol/L0.14.26.9<0.38s, p, csf
TryptophanTrpEssential aromatic amino acid630540–90/2–7 µmol/L43.13.90.51s, p, csf
Xanthurenic acidXATrp metabolite56992–35/<0.5 nmol/L0., p, csf
Folate species and catabolites.

Sample volume required: 60 µL

All concentrations are reported in nmol/L.

5-Methyl-tetrahydrofolatemTHFFolate439234>7.50.075-94-5s, p
5-Formyl-tetrahydrofolatefTHFFolate60060-20.528-96-11s, p
Folic acidFAFolate60370-20.277-86-12s, p
Serum folate as pABG equivalentsS pABGFolate>7.5s, p
RBC folate as pABG equivalentsE pABGFolate>340e
Para-aminobenzoylglutamatepABGFolate catabolite1964730-0.50.086-93-10s, p
AcetamidobenzoylglutamateapABGFolate catabolite30808680.1-10.135-65-9s, p
4-α-hydroxy-5-methyl-THFhmTHFOxidized folate0.26-106-7s, p

Folate (B9) and cobalamin (B12).

Sample volume required: 15-35 µL
Microbiological assay

Serum folates-FolateB vitamin>7.5 nmol/L2450.56s, p
Erythrocyte folatee-FolateB vitamin>340 nmol/Le
Serum cobalamins-B12B vitamin>150 pmol/L30450.82s, p
Protein biomarkers and variants.

Sample volume required: 30 µL

C-reactive proteinCRPProteinP0274110-10 µg/mL***0.14-57-90.5-0.7s, p
Serum amyloid ASAAProteinP0DJI8, P0DJI9180-20 µg/mL0.054-58-100.4-0.7s, p
CalprotectinS100AProteinP05109, P067026*0-30 µg/mL0.024-77-80.3-0.5s, p, csf
Cystatin CCnCProteinP010345*0-8 µg/mL0.014-54-50.7s, p, csf
Glycated hemoglobinHbA1cProteinP69905, F8W6P54-14 %2-6w

*Mass interference of S100A and CnC proteoforms; **ICCs of proteoforms are reported here; *** high sensitive CRP (hs-CRP)

Lipid-soluble vitamins.

Sample volume required: 50-100 µL

All-trans retinolVit. AA vitamer4453540.3-8 µmol/L, p
25-hydroxy vitamin D225OH-D2D vitamer57101480-60 nmol/L56.68.1s, p
25-hydroxy vitamin D325OH-D3D vitamer528373110-280 nmol/L3., p
α-tocopherolaTocE vitamer1498511-106 µmol/L, p
γ-tocopherolgTocE vitamer927290.5-11 µmol/L 0.13.75s, p
PhylloquinoneVit. K1K vitamer52846070.5-20 nmol/L0., p
Short-, medium- and long-chain acylcarnitines.

Sample volume required: 60 µL

All concentrations are reported in µmol/L. Read more about acylcarnitines here.

ButyrobetaineBBQuarternary ammonium7250.2-, p
CarnitineC0Carnitine2887–900.502.72.40.69s, p
AcetylcarnitineC2Acylcarnitine70457671–350., p
PropionylcarnitineC3Acylcarnitine1077380.1-, p
MalonylcarnitineC3DCAcylcarnitine918256060-, p
ButyrylcarnitineC4Acylcarnitine2131440.04-1.20.0053.43.1s, p
HydroxybutyrylcarnitineC4OHAcylcarnitine906598850.01-, p
Succinylcarnitine / MethylmalonylcarnitineC4DCAcylcarnitine1318020750.01-10.018.412.7s, p
IsovalerylcarnitineiC5Acylcarnitine64268510.04-, p
GlutarylcarnitineC5DCAcylcarnitine714644880.01-, p
Methylcrotonylcarnitine / TiglylcarnitineC5:1Acylcarnitine717502540.005-0.10.0055.03.7s, p
HexanoylcarnitineC6Acylcarnitine64268530.005-2.10.0055.63.8s, p
OctanoylcarnitineC8Acylcarnitine1237010.03-1.10.0053.53.70.41s, p
DecanoylcarnitineC10Acylcarnitine102451900.03-2.10.0053.21.7s, p
DodecanoylcarnitineC12Acylcarnitine1683810.01-, p
Tetradecanoylcarnitine (Myristoylcarnitine)C14Acylcarnitine64268540.015-0.30.0155.42.70.38s, p
HydroxytetradecanoylcarnitineC14OHAcylcarnitine714645410.005-0.080.0057.64.1s, p
Hexadecanoylcarnitine (Palmitoylcarnitine)C16Acylcarnitine4610.04-, p
HydroxyhexadecanoylcarnitineC16OHAcylcarnitine1264562280-0.050.0055.24.1s, p
Octadecanoylcarnitine (Stearoylcarnitine)C18Acylcarnitine529220560.01-0.50.0065.06.60.4s, p
HydroxyoctadecanoylcarnitineC18OHAcylcarnitine714645600.019.94.6s, p
Octadecenoylcarnitine (Oleylcarnitine)C18:1Acylcarnitine469079330.02-, p
Octadecadienylcarnitine (Linoleylcarnitine)C18:2Acylcarnitine64500150.02-, p

*Multianalyte determination is carried out in a single aliquot. Volume requirement is independent of the number of metabolites. Reported values are not strictly normal reference range, which depends on the population, and several factors, including age, gender, ethnicity and procedures for sample handling and analytical technology. Reported values are concentrations that we and others have observed or reported in various cohorts of healthy subjects. For some biomarkers the assay performance may be different from those given in published articles. This is due to continuous improvements of the methods. LODs, CVs, and ICCs are not available for all biomarkers.

May 14, 2024
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Amino acids, amino acid catabolites, acylcarnitines, TCA metabolites, ketone bodies, AGEs

Kynurenines, ratio-derived metabolites, proteins

SCFAs, indoles, choline metabolites, amino acid derived

B-vitamins, functional markers, and methyl donors, fat-soluble vitamins, essential amino acids, meat & fish intake, tobacco use & coffee intake
Biomarkers (n)

Amino acids: Alanine, Arginine, Asparagine, Aspartic acid, Glutamic acid, Glutamine, Glycine, Histidine, Isoleucine, Kynurenine, Leucine, Lysine, Methionine, Ornithine, Phenylalanine, Proline, Sarcosine, Serine, Threonine, Total cysteine, Tryptophan, Tyrosine, Valine

Amino acid catabolites: 2-Aminoadipic acid, 2-Hydroxybutyrate, 3-Hydroxysiobutyrate, α-Hydroxyglutaric acid, β-Alanine, β-Aminoisobutyrate, β-Hydroxy B-methylbutyric acid, Phenylacetylglutamine

Acylcarnities: BB, C0, C2, C3, C3-DC, C4, C4-OH, C4-DC, iC5, C5-DC, C5:1, C6, C8, C10, C12, C14, C14-OH, C16, C16-OH, C18, C18-OH, C18:1, C18:2

TCA metabolites: α-Ketoglutarate, Citrate, Fumarate, Isocitrate, Lactate, Malate, Pyruvate

Ketone bodies: Acetoacetate, 3-Hydroxybutyrate

AGEs: Carboxyethyllysine, Carboxymethyllysine


Kynurenines: 3-hydroxykynurenine, 3-hydroxyanthranilic acid, Anthranilic acid, Kynurenine, Kynurenic acid, Nicotinic acid, Nicotinamide, N1-methylnicotinamide, Picolinic acid, Quinaldic acid, Quinolinic acid, Xanthurenic acid


Proteins: C-Reactive protein, Calprotectin, Serum Amyloid A

Ratio-derived: Kynurenine/tryptophan ratio, PAr index (PLP, PL, PA)


SCFAs: Acetate, Butyrate, Formate, Isobutyrate, Isovalerate, Propionate, Valerate, α-Methylbutyrate

Indoles: 3-Indoxyl sulfate, Imidazole propionate, Indole-3-acetamide, Indole-3-acetate, Indole-3-aldehyde, Indole-3-lactate, Indole-3-propionate

Choline metabolites: Choline, Betaine, DMG, TMAO

Amino acid derived: Phenylacetylglutamine


B-vitamins, functional markers, and methyl donors: 4-Pyridoxic acid, Betaine, Choline, Cobalamin, Flavin mononucleotide, Folate, Methylmalonic acid, N1-methylnicotinamide, Nicotinamide, Nicotinic acid, Pyridoxal, Pyridoxal 5-phosphate, Pyridoxine, Riboflavin, Thiamine, Thiamine monophosphate, Total homocysteine

Fat-soluble vitamins: 25-hydroxy vitamin D2, 25-hydroxy vitamin D3, α-Tocopherol (Vit. E), All-trans retinol (Vit. A), Phylloquinone (Vit. K1), y-Tocopherol (Vit. E)

Essential amino acids: Histidine, Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Threonine, Tryptophan, Valine

Meat and fish intake: 1-Methylhistidine, 3-Methylhistidine, B-Alanine, Creatine, Creatinine, TMAO

Tobacco use and coffee intake: Cotinine, Trans-3-hydroxycotinine, Trigonelline

Volume (μl)
Analytical techniques
GC- and LC-MS/MS
GC- and LC-MS/MS
GC- and LC-MS/MS
Turnaround time (weeks)

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