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BEVITAL AS

Methods

Multianalyte determination is carried out in a single aliquot. Volume requirement is independent of the number of metabolites. ”Reported concentrations” 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 concentrations” are blood levels that we and others have observed or reported for serum/plasma in various cohorts. 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.

Analytical platforms

Short-chain fatty acids.
GC-MS/MS: 50 µL vol.
All concentrations are reported as µmol/L.
MetabolitesAbbrCategoryPubChemReportedMatrix
AcetateAceShort-chain fatty acid175-1765-300s, p
ButyrateButShort-chain fatty acid2641-30s, p
FormateFrmShort-chain fatty acid2848-100s, p, csf
IsobutyrateiButShort-chain fatty acid65900.7-6s, p, csf
IsovalerateiVlrShort-chain fatty acid104300.3-3s, p
PropionateProShort-chain fatty acid10421-13s, p
ValerateVlrShort-chain fatty acid79910.2-1s, p
α-MethylbutyrateaMBShort-chain fatty acid83140.3-3s, p
Amino- and carboxylic acids. TCA metabolites and intermediates.
GC-MS/MS: 50 µL vol.
All concentrations are reported as µmol/L.
MetabolitesAbbrCategoryPubChemReportedLODCV WtnCV BtwICCMatrixMethod
Methylmalonic acidMMACarboxylic acid487<0.260.032.430.81s, p
Total homocysteinetHcySulfur amino acid915525-150.11.42.70.72s, 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
CystathionineCystaThioether4392580.07-0.550.022.32.90.63s, p
Sarcosine*SarcMethylated amino acid10880.7-2.30.0351.31.60.68s, p
HistidineHisEssential amino acid627440-1501230.44as, p
TryptophanTrpEssential aromatic amino acid630540-900.11.62.60.51s, p
KynurenineKynTrp metabolite1611661.0-3.30.121.92.30.68s, 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 acid5962120-2901.21.41.90.54as, p
AlanineAlaHydrophobic amino acid5950230-51011.21.40.43as, p
PhenylalaninePheEssential aromatic amino acid614026-850.51.530.44as, p
IsoleucineIleEssential BCAA**630640-1400.51.51.80.53as, p
LeucineLeuEssential BCAA**610670-1700.53.440.52as, p
ProlineProHydrophobic amino acid145742110-3600.52.83.70.71as, p
ValineValEssential BCAA**6287150-3500.52.22.70.38as, p
AsparagineAsnPolar amino acid626720-1300.6330.40as, p
GlutamineGlnPolar amino acid5961390-700102.12.70.42as, p
ThreonineThrEssential amino acid628870-2401.22.43.30.44as, p
TyrosineTyrAromatic amino acid605740-1100.52.12.80.55as, p
α-Hydroxyglutaric acidaHGα-Hydroxy acid430.15-20.104.27.0s, p
3-Hydroxyisobutyrate3HIBVal metabolite44087315-2532.42.9s, p
2-HydroxybutyrateaHBα-Hydroxy acid1126610-6031.72.3s, p
3-HydroxybutyratebHBKeton body44120-70031.93.4s, p
AcetoacetateAcAcKeton body9610-200555s, p
CarboxymethyllysineCMLan AGE***1238000.05-0.30.034.34.5s, p
CarboxyethyllysineCELan AGE***234007790.05-0.40.034.14.4s, p
2-Aminoadipic acid2AAALys metabolite4690.3-2.00.24.86.9s, p
β-AminoisobutyrateBAIBAVal metabolite649560.3-2.00.22.53.7s, 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 acid4449720.5-40.21.75.2s, p, csf
MalateMalβ-Hydroxy acid5252-210.42.62.6s, 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. Short- and medium-chain acylcarnitines.
LC-MS/MS: 35 µL vol.
All concentrations are reported as µmol/L.
MetabolitesAbbrCategoryPubChemReportedLODCV WtnCV BtwICCMatrixMethod
CholineCholQuarternary ammonium3055–150.52.83.20.36s, p, u
Trimethylamine N-oxideTMAOAmine oxide11450.5–600.144.1<0.38s, p, u
BetaineBetMethylated amino acid24715–600.53.84.50.65s, p, u
DimethylglycineDMGMethylated amino acid6731.5–50.255.55.90.64s, p, u
Total homocysteinetHcyAmino acid91552<150.1120.72s, p, u
CreatineCrnα-Amino acid58610–10013.94s, p, u
CreatinineCreatα-Amino acid58850–1000.252.63.10.79s, p, u
MethionineMetEssential amino acid613718–5014.14.50.33s, p
Methionine sulfoxideMetSoPolar amino acid1589800.5–50.067.17.90.36s, p
Total cysteinetCysSulfur amino acid5862150–4000.1260.56s, p
HistidineHisEssential amino acid627440–2000.253.53.8s, p
3-Methylhistidine3-MHAmino acid921052.7–100.64.44.9s, p, u
1-Methylhistidine1-MHAmino acid649690.5–150.253.73.9s, p, u
CitrullineCtnAmino acid975010–6047-86-9s, p
ArginineArgCharged amino acid632210–1000.252.630.53s, p
Asymmetric dimethylarginineADMAGuanidinated amino acid1238310.4–10.085.25.50.53s, p
Symmetric dimethylarginineSDMAGuanidinated amino acid1691480.3–0.90.084.85.20.62s, p
HomoargininehArgGuanidinated amino acid90851–60.15.860.65s, p
TrimethyllysineTMLMethylated amino acid4401210.3–20.244.40.15s, p
ButyrobetaineBBQuarternary ammonium7250.4–20.2s, p
CarnitineC0Carnitine28820–800.50.69as, p
Carnitine, total**tC0Carnitines30–900.5s, p
AcetylcarnitineC2Acylcarnitine70457673–150.10.63s, p
PropionylcarnitineC3Acylcarnitine1077380.2–.50.050.62s, p
ButyrylcarnitineC4Acylcarnitine2131440.1–0.50.012s, p
IsovalerylcarnitineiC5Acylcarnitine64268510.06–0.20.050.68s, p
GlutarylcarnitineC5DCAcylcarnitine71464488<0.050.01s, p
HexanoylcarnitineC6Acylcarnitine64268530.01–1.20.01s, p
OctanoylcarnitineC8Acylcarnitine1237010.06–0.30.050.41s, p
DecanoylcarnitineC10Acylcarnitine102451900.2–10.02s, p
DodecanoylcarnitineC12Acylcarnitine168381<0.40.02s, p
*Measured in a separate assay after sample treatment with phospholipase D that does not affect the other metabolites of platform C, which are obtainable; **Measured in a separate assay involving alkaline hydrolysis of acylcarnitines. 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
Kynurenine pathway and B vitamins (B1, B2, B3, B6).
LC-MS/MS: 60 µL vol.
All concentrations are reported as µmol/L or nmol/L.
MetabolitesAbbrCategoryPubChemReportedLODCV WtnCV BtwICCMatrixMethod
Pyridoxal 5-phosphatePLPB6 vitamer105115–150 nmol//L0.23.55.70.7s, p
PLP7–70 nmolcsf
PyridoxalPLB6 vitamer10505–150 nmol/L0.23.55.10.62s, p
PL10–50 nmol/Lcsf
4-Pyridoxic acidPAB6 vitamer672310–200 nmol/L0.53.65.80.58s, p
PA0–15 nmol/Lcsf
PyridoxinePNB6 vitamer1054Exogenous metabolite, reflecting B vitamin supplement use14.55.2s, p
ThiamineThiB1 vitamer11302–50 nmol/L0.53.54.9s, p
Thi5–40 nmol/Lcsf
Thiamine monophosphateTMPB1 vitamer11312–15 nmol/L0.57.111.6s, p
TMP25–75 µmol/Lcsf
RiboflavinRiboB2 vitamer4935705–100 nmol/L0.267.10.79s, p
Ribo5–25 nmol/Lcsf
Flavin mononucleotideFMNB2 vitamer6439763–30 nmol/L0.45.78.10.69s, p
FMN1–2 nmol/Lcsf
CystathionineCystaThioether43925870–550 nmol/L0.023.44.50.63s, p
NeopterinNeoptPteridine44555–50 nmol/L0.75.36.60.6s, p
Neopt5–50 nmol/Lcsf
TryptophanTrpEssential aromatic amino acid630540–90 µmol/L43.13.90.51s, p
Trp2–7 µmol/Lcsf
KynurenineKynTrp metabolite1611661.0–3.3 µmol/L0.0073.65.10.68s, p
Kyn0.02–0.12 µmol/Lcsf
Kynurenic acidKATrp metabolite384520–100 nmol/L0.44.45.40.68s, p
KA0.5–15 nmol/Lcsf
Quinaldic acidQldTrp (KA) metabolite71240.2–100 nmol/L0.56-76-8s, p
Qld0.1–10 nmol/Lcsf
Anthranilic acidAATrp metabolite2277–30 nmol/L27.49.50.64s, p
AA3–14 nmol/Lcsf
3-HydroxykynurenineHKTrp metabolite1181125–80 nmol/L23.24.50.62s, p
HK0.1–25 nmol/Lcsf
Xanthurenic acidXATrp metabolite56992–35 nmol/L0.54.85.70.58s, p
XA<0.5 nmol/Lcsf
3-Hydroxyanthranilic acidHAATrp metabolite8610–80 nmol/L24.660.54s, p
HAA0–7 nmol/Lcsf
Picolinic acidPicTrp metabolite101820–100 nmol/L86-75-8s, p
Pic5–100 nmol/Lcsf
Quinolinic acidQATrp metabolite1066150–700 nmol/L1.67-87-10s, p
QA10–150 nmol/Lcsf
Nicotinic acidNAB3 vitamer938<20 nmol/L1045.4s, p
NicotinamideNAMB3 vitamer936100–600 nmol/L204.26.1s, p
NAM5–200 nmol/Lcsf
N1-methylnicotinamidemNAMB3 vitamer45720–250 nmol/L545.5s, p
mNAM5–100 nmol/Lcsf
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, u
Trans-3-hydroxycotinineOHCotNicotine metabolite107963Concentrations relative to cotinine, with OHCot/cotininine ratio ranging from 0.02-0.9023.64.50.85s, p, u
TrigonellineTrigAlkaloid55700.4–25 µmol/L0.012.93.80.66s, p
Trimethylamine N-oxideTMAOAmine oxide11450.5–60 µmol/L0.14.26.9<0.38s, p, u
TMAO0.2–12 µmol/Lcsf
SuccinateSucDicarboxylic acid11102–12 µmol/Ls, p
Imidazole propionateImPHistidine metabolite706303–500 nmol/L2s, p
3-Indoxyl sulfate3ISIndole, Trp metabolite102580.9–30 µmol/L0.2s, 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
Folate species and catabolites.
LC-MS/MS: 60 µL vol.
All concentrations are reported as nmol/L.
MetabolitesAbbrCategoryPubChemReportedLODCV WtnCV BtwMatrixMethod
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).
Microbiological assay: 15-35 µL vol.
All concentrations are reported as nmol/L or pmol/L.
MetabolitesAbbrCategoryReportedLODCV WtnCV BtwICCMatrixMethod
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.
MALDI-TOF MS: 30 µL vol.

All concentrations are reported as µmol/L or %.

ProteinsAbbrCategoryUniProtProteoformsReportedLODCV WtnCV BtwICC**MatrixMethod
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 and long-chain acylcarnitines.
LC-MS/MS: 50-100 µL vol.
All concentrations are reported as µmol/L or nmol/L.
MetabolitesAbbrCategoryPubChemReportedLODCV WtnCV BtwICCMatrixMethod
All-trans retinolVit. AA vitamer4453540,3-8 µmol/L 0.12.93.60.87s, p
25-hydroxy vitamin D225OH-D2D vitamer57101480-60 nmol/L56.68.1s, p
25-hydroxy vitamin D325OH-D3D vitamer528373110-280 nmol/L3.35.16.10.71s, p
α-tocopherolaTocE vitamer1498511-106 µmol/L 24.35.80.86s, p
γ-tocopherolgTocE vitamer927290,5-11 µmol/L 0.13.75s, p
PhylloquinoneVit. K1K vitamer52846070,5-20 nmol/L0.54.87.40.49s, p
MyristoylcarnitineC14Acylcarnitine642685410-216 nmol/L0.38as, p
PalmitoylcarnitineC16Acylcarnitine46140-310 nmol/L0.49as, p
StearoylcarnitineC18Acylcarnitine5292205617-100 nmol/L0.40as, p
OleoylcarnitineC18:1Acylcarnitine4690793340-412 nmol/L0.66bs, p
LinoleylcarnitineC18:2Acylcarnitine645001510-297 nmol/Ls, 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 bXiaofei 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.
This platform is under development: Short-, medium- and long-chain acylcarnitines.
LC (HILIC)-MS/MS
All concentrations are reported as µmol/L.
MetabolitesAbbrCategoryPubChemReportedLODICCMatrix
ButyrobetaineBBQuarternary ammonium7250.4-20.2s, p
CarnitineC0Carnitine28820-800.50.76bs, p
AcetylcarnitineC2Acylcarnitine70457673-150.10.63s, p
PropionylcarnitineC3Acylcarnitine1077380.2-1.50.050.62s, p
ButyrylcarnitineC4Acylcarnitine2131440.1-0.50.0120.79bs, p
IsovalerylcarnitineiC5Acylcarnitine64268510.06-0.20.050.68s, p
HexanoylcarnitineC6Acylcarnitine64268530.01-1.20.01s, p
OctanoylcarnitineC8Acylcarnitine1237010.06-0.30.050.41s, p
DecanoylcarnitineC10Acylcarnitine102451900.2-10.020.67bs, p
DodecanoylcarnitineC12Acylcarnitine168381< 0.40.02s, p
MyristoylcarnitineC14Acylcarnitine64268540.01-0.10.38as, p
PalmitoylcarnitineC16Acylcarnitine4610.05-0.250.49as, p
StearoylcarnitineC18Acylcarnitine529220560.03-0.10.40as, p
OleoylcarnitineC18:1Acylcarnitine469079330.04-0.2s, p
LinoleylcarnitineC18:2Acylcarnitine64500150.01-0.1s, 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 bXiaofei 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.
November 21, 2022

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