Methods
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
- A
- B
- C
- D
- E
- F
- G
- H
- I
Short-chain fatty acids.
Sample volume required: 50 µL
GC-MS/MS
All concentrations are reported in µmol/L.
Amino- and carboxylic acids. TCA metabolites and intermediates.
Sample volume required: 50 µL
GC-MS/MS
All concentrations are reported in µmol/L.
*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
LC-MS/MS
All concentrations are reported in µmol/L.
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
LC-MS/MS
All concentrations are reported in µmol/L or nmol/L.
Folate species and catabolites.
Sample volume required: 60 µL
LC-MS/MS
All concentrations are reported in nmol/L.
Folate (B9) and cobalamin (B12).
Sample volume required: 15-35 µL
Microbiological assay
All concentrations are reported in nmol/L or pmol/L.
Serum folate | s-Folate | B vitamin | >7.5 nmol/L | 2 | 4 | 5 | 0.56 | s, p | |
Erythrocyte folate | e-Folate | B vitamin | >340 nmol/L | e | |||||
Serum cobalamin | s-B12 | B vitamin | >150 pmol/L | 30 | 4 | 5 | 0.82 | s, p |
Protein biomarkers and variants.
Sample volume required: 30 µL
MALDI-TOF MS
All concentrations are reported in µg/ml or %.
*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
LC-MS/MS
All concentrations are reported in µmol/L or nmol/L.
Short-, medium- and long-chain acylcarnitines.
Sample volume required: 60 µL
LC (HILIC)-MS/MS
All concentrations are reported in µmol/L. Read more about acylcarnitines here.
Butyrobetaine | BB | Quarternary ammonium | 725 | 0.2-2 | 0.01 | 4.3 | 2.5 | s, p | |
Carnitine | C0 | Carnitine | 288 | 7–90 | 0.50 | 2.7 | 2.4 | 0.69 | s, p |
Acetylcarnitine | C2 | Acylcarnitine | 7045767 | 1–35 | 0.06 | 3.0 | 2.1 | 0.63 | s, p |
Propionylcarnitine | C3 | Acylcarnitine | 107738 | 0.1-1.5 | 0.01 | 3.8 | 1.6 | 0.62 | s, p |
Malonylcarnitine | C3DC | Acylcarnitine | 91825606 | 0-0.5 | 0.05 | 2.8 | 2.4 | s, p | |
Butyrylcarnitine | C4 | Acylcarnitine | 213144 | 0.04-1.2 | 0.005 | 3.4 | 3.1 | s, p | |
Hydroxybutyrylcarnitine | C4OH | Acylcarnitine | 90659885 | 0.01-0.5 | 0.01 | 5.3 | 1.6 | s, p | |
Succinylcarnitine / Methylmalonylcarnitine | C4DC | Acylcarnitine | 131802075 | 0.01-1 | 0.01 | 8.4 | 12.7 | s, p | |
Isovalerylcarnitine | iC5 | Acylcarnitine | 6426851 | 0.04-0.5 | 0.01 | 2.7 | 1.8 | 0.68 | s, p |
Glutarylcarnitine | C5DC | Acylcarnitine | 71464488 | 0.01-0.3 | 0.01 | 8.0 | 8.2 | s, p | |
Methylcrotonylcarnitine / Tiglylcarnitine | C5:1 | Acylcarnitine | 71750254 | 0.005-0.1 | 0.005 | 5.0 | 3.7 | s, p | |
Hexanoylcarnitine | C6 | Acylcarnitine | 6426853 | 0.005-2.1 | 0.005 | 5.6 | 3.8 | s, p | |
Octanoylcarnitine | C8 | Acylcarnitine | 123701 | 0.03-1.1 | 0.005 | 3.5 | 3.7 | 0.41 | s, p |
Decanoylcarnitine | C10 | Acylcarnitine | 10245190 | 0.03-2.1 | 0.005 | 3.2 | 1.7 | s, p | |
Dodecanoylcarnitine | C12 | Acylcarnitine | 168381 | 0.01-0.5 | 0.01 | 2.3 | 1.4 | s, p | |
Tetradecanoylcarnitine (Myristoylcarnitine) | C14 | Acylcarnitine | 6426854 | 0.015-0.3 | 0.015 | 5.4 | 2.7 | 0.38 | s, p |
Hydroxytetradecanoylcarnitine | C14OH | Acylcarnitine | 71464541 | 0.005-0.08 | 0.005 | 7.6 | 4.1 | s, p | |
Hexadecanoylcarnitine (Palmitoylcarnitine) | C16 | Acylcarnitine | 461 | 0.04-1.3 | 0.01 | 3.1 | 2.6 | 0.49 | s, p |
Hydroxyhexadecanoylcarnitine | C16OH | Acylcarnitine | 126456228 | 0-0.05 | 0.005 | 5.2 | 4.1 | s, p | |
Octadecanoylcarnitine (Stearoylcarnitine) | C18 | Acylcarnitine | 52922056 | 0.01-0.5 | 0.006 | 5.0 | 6.6 | 0.4 | s, p |
Hydroxyoctadecanoylcarnitine | C18OH | Acylcarnitine | 71464560 | 0.01 | 9.9 | 4.6 | s, p | ||
Octadecenoylcarnitine (Oleylcarnitine) | C18:1 | Acylcarnitine | 46907933 | 0.02-1 | 0.01 | 3.4 | 2.2 | 0.66 | s, p |
Octadecadienylcarnitine (Linoleylcarnitine) | C18:2 | Acylcarnitine | 6450015 | 0.02-0.3 | 0.01 | 3.5 | 2.4 | s, p |
Short-chain fatty acids.
Volume: 50 µL
All concentrations are reported as µmol/L; CVs (%) shown as within-day (Wtn-day) and between-day (Btw-day) variations; sample matrices: serum (s), plasma (p), cerebrospinal fluid (csf).
Amino- and carboxylic acids. TCA metabolites and intermediates. Volume: 50 µL
All concentrations are reported as µmol/L; CVs (%) shown as within-day (Wtn-day) and between-day (Btw-day) variations; sample matrices: serum (s), plasma (p), urine (u).
*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.
Volume: 35 µL
All concentrations are reported as µmol/L; CVs (%) shown as within-day (Wtn-day) and between-day (Btw-day) variations; sample matrices: serum (s), plasma (p), urine (u).
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).
Volume: 60 µL
CVs (%) shown as within-day (Wtn-day) and between-day (Btw-day) variations; sample matrices: serum (s), plasma (p), urine (u), cerebrospinal fluid (csf).
Folate species and catabolites.
LC-MS/MS: 60 µL vol.
All concentrations are reported as nmol/L. CVs (%) shown as within-day (Wtn-day) and between-day (Btw-day) variations; sample matrices: serum (s), plasma (p).
Folate (B9) and cobalamin (B12).
Microbiological assay: 15-35 µL vol.
Serum folate | s-Folate | B vitamin | >7.5 nmol/L | 2 | 4 | 5 | 0.56 | s, p | |
Erythrocyte folate | e-Folate | B vitamin | >340 nmol/L | e | |||||
Serum cobalamin | s-B12 | B vitamin | >150 pmol/L | 30 | 4 | 5 | 0.82 | s, p |
Protein biomarkers and variants.
MALDI-TOF MS: 30 µL vol.
*Mass interference of S100A and CnC proteoforms
**ICCs of proteoforms can be found here
*** high sensitive CRP (hs-CRP)
Lipid-soluble vitamins.
LC-MS/MS: 50-100 µL vol.
Short-, medium- and long-chain acylcarnitines.
LC (HILIC)-MS/MS.
All concentrations are reported as µmol/L.
Butyrobetaine | BB | Quarternary ammonium | 725 | 0.2-2 | 0.01 | 4.3 | 2.5 | s, p | |
Carnitine | C0 | Carnitine | 288 | 7–90 | 0.50 | 2.7 | 2.4 | 0.69 | s, p |
Acetylcarnitine | C2 | Acylcarnitine | 7045767 | 1–35 | 0.06 | 3.0 | 2.1 | 0.63 | s, p |
Propionylcarnitine | C3 | Acylcarnitine | 107738 | 0.1-1.5 | 0.01 | 3.8 | 1.6 | 0.62 | s, p |
Malonylcarnitine | C3DC | Acylcarnitine | 91825606 | 0-0.5 | 0.05 | 2.8 | 2.4 | s, p | |
Butyrylcarnitine | C4 | Acylcarnitine | 213144 | 0.04-1.2 | 0.005 | 3.4 | 3.1 | s, p | |
Hydroxybutyrylcarnitine | C4OH | Acylcarnitine | 90659885 | 0.01-0.5 | 0.01 | 5.3 | 1.6 | s, p | |
Succinylcarnitine / Methylmalonylcarnitine | C4DC | Acylcarnitine | 131802075 | 0.01-1 | 0.01 | 8.4 | 12.7 | s, p | |
Isovalerylcarnitine | iC5 | Acylcarnitine | 6426851 | 0.04-0.5 | 0.01 | 2.7 | 1.8 | 0.68 | s, p |
Glutarylcarnitine | C5DC | Acylcarnitine | 71464488 | 0.01-0.3 | 0.01 | 8.0 | 8.2 | s, p | |
Methylcrotonylcarnitine / Tiglylcarnitine | C5:1 | Acylcarnitine | 71750254 | 0.005-0.1 | 0.005 | 5.0 | 3.7 | s, p | |
Hexanoylcarnitine | C6 | Acylcarnitine | 6426853 | 0.005-2.1 | 0.005 | 5.6 | 3.8 | s, p | |
Octanoylcarnitine | C8 | Acylcarnitine | 123701 | 0.03-1.1 | 0.005 | 3.5 | 3.7 | 0.41 | s, p |
Decanoylcarnitine | C10 | Acylcarnitine | 10245190 | 0.03-2.1 | 0.005 | 3.2 | 1.7 | s, p | |
Dodecanoylcarnitine | C12 | Acylcarnitine | 168381 | 0.01-0.5 | 0.01 | 2.3 | 1.4 | s, p | |
Tetradecanoylcarnitine (Myristoylcarnitine) | C14 | Acylcarnitine | 6426854 | 0.015-0.3 | 0.015 | 5.4 | 2.7 | 0.38 | s, p |
Hydroxytetradecanoylcarnitine | C14OH | Acylcarnitine | 71464541 | 0.005-0.08 | 0.005 | 7.6 | 4.1 | s, p | |
Hexadecanoylcarnitine (Palmitoylcarnitine) | C16 | Acylcarnitine | 461 | 0.04-1.3 | 0.01 | 3.1 | 2.6 | 0.49 | s, p |
Hydroxyhexadecanoylcarnitine | C16OH | Acylcarnitine | 126456228 | 0-0.05 | 0.005 | 5.2 | 4.1 | s, p | |
Octadecanoylcarnitine (Stearoylcarnitine) | C18 | Acylcarnitine | 52922056 | 0.01-0.5 | 0.006 | 5.0 | 6.6 | 0.4 | s, p |
Hydroxyoctadecanoylcarnitine | C18OH | Acylcarnitine | 71464560 | 0.01 | 9.9 | 4.6 | s, p | ||
Octadecenoylcarnitine (Oleylcarnitine) | C18:1 | Acylcarnitine | 46907933 | 0.02-1 | 0.01 | 3.4 | 2.2 | 0.66 | s, p |
Octadecadienylcarnitine (Linoleylcarnitine) | C18:2 | Acylcarnitine | 6450015 | 0.02-0.3 | 0.01 | 3.5 | 2.4 | s, p |
*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 values 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.