Targeted methods can provide semiquantitative “fold change” information or full quantitative measurement of the target compounds. Overall targeted approaches provide the advantage that the detection/quantification and validation of the predefined analytes is better controlled. LC-MS/MS is the most widely applied platform for targeted profiling. An increasing number of targeted methods are appearing in the international literature
The Biomic group has developed targeted metabolomics that provide coverage and has applied these methods in the analysis of several thousands of samples of various types (blood, urine, amniotic fluid, tissue, feces, cell culture but also foods such as grape, honey, royal jelly, wine, olive oil and other specimens). Metabolites covered include aminoacids, sugars, nucleotides, vitamins, lipids cofactors and others.
We provide results that can range from semi-quantitative approach (fold change) to absolute quantification using isotopically labelled internal standards.
Amino acids can be found in biological fluids and have key roles in many biological processes, such as protein synthesis, neurotransmission, and the regulation of metabolic pathways. Characteristically, branched-chain amino acids (BCAAs) (valine (Val), leucine (Leu), and isoleucine (Ile)) are involved in protein synthesis, while tyrosine (Tyr) and Phenylalanine (Phe) are intermediate metabolites in the biosynthesis of catecholamines. Also, γ-aminobutyric acid (GABA), glutamic acid (Glu), and taurine (Tau), act as neurotransmitters that modulate transmission in the synaptic cleft. It has been shown that alterations in levels of amino acids in biological fluids are related to many types of diseases such as diabetes, liver disease, kidney disease and different types of cancers. Therefore, there is a great need for the development of analytical methods that can detect these changes in levels of amino acids in biological samples (e.g. plasma, urine, and sweat) which subsequently could be used as biomarkers for early diagnosis and elucidation of mechanisms of diseases.
Our HILIC-MS/MS method covers all major amino acid providing unsurpassed sensitivity, specificity, wide dynamic range, robustness, and reproducibility. The method has been applied in a number of studies and matrices (CSF, blood, various other biological fluids and extracts).
Virgiliou, C., Sampsonidis, I., Gika, H.G., Raikos, N., Theodoridis, G.A. Development and validation of a HILIC-MS/MS multitargeted method for metabolomics applications (2015) Electrophoresis, 36 (18), pp. 2215-2225.
|Deoxycholic acid||Taurocholic acid|
|taurodeoxycholic acid||Cholic acid|
|Chenodeoxycholic acid||Glycodeoxycholic acid|
|Dehydrocholic acid||tauroursodeoxycholic acid|
|Lithocholic acid||Taurochenodeoxycholic acid|
|Glycocholic acid||Ursodeoxycholic acid|
Bile acids (BAs) are important end products of cholesterol metabolism. The main function of bile acids is the absorption of fats and fat-soluble vitamins. In addition, they are important for the maintenance of cholesterol homeostasis, excretion of toxic substances, induction of bile flow, acting as signalling molecules and intermediates between host and gut microbiome, ultimately influencing glucose homeostasis, lipid metabolism and energy expenditure. BAs are primarily synthesized from cholesterol in the liver and play house keeping roles in many physiological processes as described above. BAs are modulated by gut microbiome, and dysregulation of the homeostasis of circulating BAs pool is associated with liver and intestinal diseases, as well as neuropsychiatric symptoms.
In humans BAs comprise primary BAs synthesized in the liver (including cholic acid (CA) and chenodeoxycholic acid (CDCA)), secondary BAs produced in the gut (deoxycholic acid (DCA), lithocholic acid (LCA), and ursodeoxycholic acid (UDCA)) generated by deconjugation and dehydroxylation by intestinal bacteria, and tertiary BAs which are synthesized in both the liver and gut via modification of secondary BAs, such as sulfation, glucuronidation, and glucosidation. BAs are reabsorbed and transported back to the liver in a process known as enterohepatic circulation. BAs biotransformation is a complex process resulting in structural diversity of molecular species of BAs. Also, BAs present an extended concentration range in human biofluids and between physiological and disease phenotype (0.1-2000 nM). Thus, considering the biological significance of Bas and their complex profile, there is an apparent need for high quality quantitative results in various biospecimens and clinical applications.
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is considered as the method of choice for the quantification of BAs and their conjugates.
|N- Palmitoyl-D-erythro-sphingosine (Cer(d18:1/16:0))|
|N- stearoyl- D- erythro- sphingosine (Cer(d18:1/18:0))|
|N-nervonoyl-D- erythro-sphingosine (Cer(d18:1/24:1))|
|Alpha Linolenic (18:3n3)||Vaccenic (18:1n7)|
|Docosapentaenoic (22:5n3)||Oleic (18:1n9)|
|Eicosapentaenoic (20:5n3)||11-Eicosenoic (20:1n9)|
|Docosahexaenoic (22:6n3)||Nervonic (24:1n9)|
|Linoleic (18:2n6)||Capric (10:0)|
|Gamma Linolenic (18:3n6)||Lauric (12:0)|
|Eicosadienoic (20:2n6)||Myristic (14:0)|
|Dihomogamma Linolenic (20:3n6)||Palmitic (16:0)|
|Arachidonic (20:4n6)||Stearic (18:0)|
|Docosadienoic (22:2n6)||Arachidic (20:0)|
|Docosatetraenoic (22:4n6)||Behenic (22:0)|
|Myristoleic (14:1n5)||Lignoceric (24:0)|
|Pyruvic acid||3-Methylglutaric||Tartaric acid|
Vitamins are essential to maintenance of healthy homeostasis, to catalyze biochemical reactions and aid in metabolic functions in humans. Deficiency in vitamins or excess can cause health problems such as cardiovascular disease, kidney disease, depression, and anemia. Therefore, there is a great need to monitor their concentration in plasma and other biological fluids as a means of diagnosis of different diseases and evaluation of treatment therapies. The majority of methods used for determination of vitamins in biofluids are chromatographic, such as high-performance liquid chromatography (HPLC). Over the last decade, LC-MS/MS methods have been heavily used for quantification of vitamins in complex matrices (e.g. plasma, serum, urine, breast milk, or amniotic fluid) due to their high sensitivity, selectivity, wide dynamic range of concentrations, and measurement of all forms of vitamins in human biofluids.
There are two groups of vitamins: water-soluble and fat-soluble vitamins. Water-soluble vitamins include ascorbic acid (vitamin C) and B group vitamins: thiamine (B1), riboflavin (B2), niacin (B3), pantothenic acid (B5), pyridoxal (B6), folic acid (B9), and cyanocobalamin (B12) . Fat-soluble vitamins include retinol (vitamin A), tocopherol (vitamin E), radiostol (vitaminD), and vitamin K. The measurement of vitamins in human biological fluids is important for evaluation of vitamin status, disorders of lipid malabsorption, diagnosis of anemia or hypervitaminosis, toxicity assessment and various other purposes. Vitamins are also useful indicators, especially in clinical, nutritional and metabolomics studies.
Biomics group can undertake the analysis of selected vitamins or groups of vitamins in various matrices using validated methods.