Targeted Metabolomics Services


Updated 14/05/2024

What is measured?

The method includes protein precipitation by trichloroacetic acid, which oxidises 7,8-dihydroneopterin to neopterin, and measures total neopterin, which is higher than concentrations obtained using assays that measure only neopterin.
Method: LC-MS/MS (1).

What is neopterin?

Neopterin, a pyrazino-pyrimidine compound, is synthesized by monocytes and macrophages in response to interferon-𝛾 (IFN-γ) produced by activated T cells. Both total neopterin and neopterin reflect cellular immune response, because IFN-γ induces a step that precedes formation of 7,8-dihydroneopterin in the neopterin pathway. Increased concentrations are observed in infections by viruses, including human immunodeficiency virus (HIV), infections by intracellular bacteria and parasites, autoimmune diseases, malignant tumour diseases and in allograft rejection episodes. Neopterin in serum/plasma shows a strong, positive relation to total homocysteine and to the kynurenine/tryptophan ratio (KTR) (2).
Since IFN-g induce a step that precedes formation of 7,8-dihydroneopterin in the neopterin pathway, the use of neopterin and total neopterin both reflect inflammation. The use of total neopterin is further supported by its strong correlation with neopterin in urine (3) and plasma, and it has been suggested that neopterin and total neopterin are of equal value for clinical diagnosis (3).


Assessment of cellular immune response.

Specimen, collection and processing

Matrix: Plasma and serum. Neopterin increases (up to 100%) in samples with hemolysis.
Volume: Minimum volume is 60 µL, but 200 µL is optimal and allows reanalysis.
Preparation: Samples should be put on ice, and protected from exposure to ultraviolet light and sunlight.


Frozen, on dry ice and protected from exposure to ultraviolet light and sunlight. (for general instruction on transportation, click here)

Reported values, interpretation

Total neopterin correlates strongly with neopterin in urine and plasma, and with plasma KTR. Neopterin and total neopterin are of equal value for the assessment of immune activation (3).
Reported values: 5-50 nmol/L.
Intraclass correlation coefficient (ICC): 0.60.


1. Midttun, O., Hustad, S., and Ueland, P.M. (2009). Quantitative profiling of biomarkers related to B-vitamin status, tryptophan metabolism and inflammation in human plasma by liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Sp 23, 1371-79.
2. Capuron, L., Geisler, S., Kurz, K., Leblhuber, F., Sperner-Unterweger, B., and Fuchs, D. (2014). Activated immune system and inflammation in healthy ageing: Relevance for tryptophan and neopterin Metabolism. Curr Pharm Des 20, 6048-057.
3. Fuchs, D., Milstien, S., Krämer, A., Reibnegger, G., Werner, E.R., Goedert, J.J., Kaufman, S., and Wachter, H. (1989). Urinary neopterin concentrations vs total neopterins for clinical utility. Clin Chem 35, 2305-07.

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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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