In the past GC/MS was used to determine the +/- 70 organic acids that were performed.
Now we developed a special new more sensitive quantitative method based on LC-MS/MS
with tandem Mass spectrometry
allowing us to perform the tests with less sample volume in urine, increased sensitivity, reduced disturbance/errors and allowing us to perform 90 organic acids within a short time of analyzing.
Organic acid disorders (organic acidemia) are a group of (inherited) metabolic conditions. Most of the organic acidemias caused by defective autosomal genes for various enzymes are for amino acid metabolism. Neurological and physiological harms are caused by this impaired ability to synthesize a key enzyme required to break down a specific amino acid, or a group of amino acids, resulting in acidemia and toxicity to specific organs systems. Most are inherited as autosomal recessive diseases. Organic acid related disorders may affect many metabolic pathways including amino acid, lipid metabolism, fatty acid oxidation and the Krebs cycle or citric acid cycle and mitochondrial function.
In the past the test performed were mostly focused on all ages including environmental, yeast and bacterial influences. Additional are Organic acidemias usually diagnosed in infancy, characterized by urinary excretion of abnormal amounts or types of organic acids. The diagnosis is usually made by detecting an abnormal pattern of organic acids in a urine sample. In some conditions, the urine is always abnormal; in the others the characteristic substances are only present intermittently. Many of the organic acidemias are detectable by newborn screening with tandem Mass spectrometry . These disorders vary in their prognosis, from manageable to fatal, and usually affect more than one organ system, especially the central nervous system.
Main methods and procedures that have been selected are based on EN ISO 13485, 98/79/EC and CAP rules.
1.
Why do use LC/MS/MS Not GC/MS for Urine Organic acid analysis?
LC/MS/MS seems to be the gold standard in sample analysis nowadays. The majority of published peer-reviewed articles are for LC/MS/MS. Also it seems possible to add certain organic acids of your special interest. Please inform us if this is applicable to your practice.
2. New Organic Acids were Added to the old Panel.
1. 3-methylglutaric
3-methylglutaric acid is an organic acid classically associated with two distinct leucine pathway enzyme deficiencies. 3-methylglutaric acid is excreted in urine of individuals harboring deficiencies in 3-hydroxy-3-methylglutaryl (HMG) CoA lyase (HMGCL) or 3-methylglutaconyl CoA hydratase (AUH). Whereas 3MG CoA is not part of the leucine catabolic pathway, it is likely formed via a side reaction involving reduction of the α-ß trans double bond in the leucine pathway intermediate, 3-methylglutaconyl CoA. While the metabolic basis for the accumulation of 3MG acid in subjects with deficiencies in HMGCL or AUH is apparent, the occurrence of 3MG aciduria in a host of unrelated inborn errors of metabolism associated with compromised mitochondrial energy metabolism is less clear. Herein, a novel mitochondrial biosynthetic pathway termed "the acetyl CoA diversion pathway", provides an explanation. The pathway is initiated by defective electron transport chain function which, ultimately, inhibits acetyl CoA entry into the TCA cycle. When this occurs, 3MG acid is synthesized in five steps from acetyl CoA via a novel reaction sequence, providing a metabolic rationale for the connection between 3MG aciduria and compromised mitochondrial energy metabolism.
2. Methylhippuric acid
Methylhippuric acid is the result of exposure to the solvent xylene that is widespread in the environment. Xylene is found in paints, lacquers, cleaning agents, pesticides, and gasoline. It is also used in the pathology laboratory for tissue processing.
3. 3,4-Dihydroxyhydrocinnamic acid
3,4-dihydroxyphenylpropionic acid is found in red beetroot, common beet, olives, and correlated with coffee intake. One of the most abundant phenolates, formed by microbial transformation of dietary polyphenols and endogenous metabolites such as dopamine, phenylalanine, tyrosine, and tryptophan. 3,4-dihydroxyphenylpropionic acid is highly correlated with homovanillic acid (HVA). 3,4-dihydroxyphenylpropionic acid has antioxidant properties and significantly inhibited the secretion of pro-inflammatory cytokines.
4. 3-Hydroxypropionic acid
3-Hydroxypropionic acid is a carboxylic acid. It is an intermediate in the breakdown of branched-chain amino acids and propionic acid from the gut. Typically it originates from propionyl-CoA and a defect in the enzyme propionyl carboxylase. This leads to a buildup in propionyl-CoA in the mitochondria. Such a buildup can lead to a disruption of the esterified CoA:free CoA ratio and ultimately to mitochondrial toxicity. Detoxification of these metabolic end products occurs via the transfer of the propionyl moiety to carnitine-forming propionyl-carnitine, which is then transferred across the inner mitochondrial membrane. 3-Hydroxypropionic acid is then released as the free acid. As an industrial chemical, it is used in the production of various chemicals such as acrylates in industry. When present in sufficiently high levels, 3-hydroxypropionic acid can act as an acidogen and a metabotoxin. An acidogen is an acidic compound that induces acidosis, which has multiple adverse effects on many organ systems. A metabotoxin is an endogenously produced metabolite that causes adverse health effects at chronically high levels. Chronically high levels of hydroxypropionic acid are associated with many inborn errors of metabolism including biotinidase deficiency, malonic aciduria, methylmalonate semialdehyde dehydrogenase deficiency, methylmalonic aciduria, methylmalonic aciduria due to cobalamin-related disorders, and propionic acidemia. Hydroxypropionic acid is an organic acid. Abnormally high levels of organic acids in the blood (organic acidemia), urine (organic aciduria), the brain, and other tissues lead to general metabolic acidosis.
5. 3-Methylcrotonyl glycine
3-Methylcrotonyl glycine is a metabolite of carnitine that has been shown to be an effective marker for detection of creatine kinase (CK) deficiency in patients with infantile mitochondrial encephalomyopathy. 3-Methylcrotonyl glycine is used for the diagnosis of metabolic disorders such as fatty acid oxidation defects, enzyme deficiencies, and inherited metabolic disorders. 3-Methylcrotonyl glycine can be used for the diagnosis of various symptoms including muscle weakness, fatigue, developmental delay, and seizures in children.
6. 4-Hydroxybenzoic acid
The presence of organic compounds such as p-Hydroxybenzoate in the urine may point towards significant dysbiosis (=impaired microbiota). p-Hydroxybenzoat may reflect intestsinal overgrowth, usually accomanied by microbal hyperpermeability.
7. 2-Hydroxyglutaric acid
2-Hydroxyglutaric acid is related to 2-Hydroxyglutaric aciduria.
8. Alpha-ketobutyric acid
Alpha-ketobutyric acid results from the breakdown of threonine or methionine during glutathione production.
- Specifically, cystathionine is metabolized to alpha-ketobutyric acid and cysteine.
- a- ketobutyric acid enters the mitochondrial matrix and get converted to propionyl-CoA by the branched chain keto-acid dehydrogenase complex (BCKDHC) and enters the Krebs cycle at succinyl-CoA.
- Evaluate lactate and the branched chain keto acids
- Evaluate alpha-hydroxybutyric acid
- Associated Nutrients: Vitamin B3
- a -Ketobutyric acid is produced from cystine, along with hydrogen sulfide (H2S) as a by-product.
- a- Ketobutyric acid is reversibly converted to a- hydroxybutyric acid.
9. Benzoic Acid
Benzoic Acid
was one of the compounds first found to be elevated in urine from patients with intestinal bacterial overgrowth of various origins. Many people with intestinal bacterial overgrowth resulting from cystic fibrosis, unclassified enteritis, celiac disease, or short bowel syndrome have elevated benzoic Acid along with varying degrees of elevated phenylacetate, p-hydroxybenzoate, and p-hydroxyphenylacetate.
Benzoic acid, a common food component used as a preservative in packaged foods such as pickles and lunch meats, also occurs naturally in cranberries and other fruits, a factor to take into account when interpreting elevated hippurate levels in urine. Whether the source is dietary intake or jejunal bacterial metabolism, benzoate should be rapidly converted to hippurate by conjugation with glycine.
10. Formiminoglutamic acid
Formiminoglutamate (FIGLU) is a functional marker of insufficiency of folic acid, another B-vitamin, and is a compound made from the amino acid histidine.
Pregnant women especially need to have adequate folic acid to prevent birth defects. Folic acid insufficiency can play a role in childhood development problems, depression, immune function, and is a risk factor for cardiovascular disease.
Insufficiency of folic acid leads to high urinary Formiminoglutamate.
11. Hexanoyl glycine
Hexanoyl glycine is an acylated amino acid that is used as a urinary biomarker for several indications. It is normally biosynthesized from hexanoyl-CoA and glycine by the mitochondrial enzyme glycine N-acyltransferase. Increased urinary excretion of hexanoyl glycine in humans is indicative of a deficiency in medium-chain acyl-CoA dehydrogenase. Increased urinary hexanoyl glycine can also be used as a biomarker for exposure to gamma radiation. Levels of hexanyl glycine can also be elevated during cancer.
12. Isocitric acid
Citric acid, cis-aconitic acid, and isocitric acid are the first three metabolites in the Krebs Citric Acid energy production cycle, which operates in the mitochondria of your cells.
Citrate, cis-Aconitate (and Isocitrate) are involved in both energy production and removal of toxic ammonia.High levels can indicate ammonia toxicity. Chronic loss of these valuable compounds can contribute to loss of organ reserve and disturbances in neurological function. If they are low they can indicate a need for essential amino acids, especially arginine.
13. Mevalonic acid
Urinary mevalonic acid levels are used as a tool in the diagnostic process Mevalonate kinase deficiency (MKD).
14.
Phenylglyoxylic acid
Phenylglyoxylic acid (PhGA) and mandelic acid (MA) are two popular urinary markers of occupational exposure to styrene.
15. Picolinic acid
Picolinate is a neurotransmitter metabolism marker and is produced under inflammatory conditions. Picolinate is a catabolite of the amino acid tryptophan through the kynurenine pathway . Its function is unclear, but it has been implicated in a variety neuro-protective, immunological, and anti-proliferative affects. In addition, it is suggested to assist in the absorption of zinc ions and other divalent or trivalent ions through the small intestine.
Nutrient Associations:
- Antioxidants (Vit C, Vit E, Lipoic acid)
- Omega 3 fatty acids.
16. Pimelic acids
Pimelic acids are excreted in elevated amounts in urine in disorders of mitochondrial beta-oxidation and disorders of peroxisomal beta-oxidation, for which they are of significant diagnostic value. Pimelic acid originating from fatty acid synthesis pathway is a bona
fide precursor of biotin in B. subtilis.
17. Propionylglycine
The level of propionylglycine is elevated in patients with propionic acidemia, which is an inherited deficiency of propionyl-CoA carboxylase.
18. Suberyl glycine
Suberylglycine was found in the urine from a patient with C6-C10-ω-dicarboxylic aciduria and unexplained episodes of lethargy and unconsciousness.
19. Succinylacetone
Succinylacetone (SA) is used for the diagnosis and monitoring of patients with tyrosinemia type I.
20. Tiglylglycine
Tiglylglycine (TG) is associated with both mitochondrial and/or genetic disorders. Toxic chemical exposure may be one of the most common causes of mitochondrial dysfunction. In mitochondrial disorders of the respiratory chain, TG values are usually more moderately increased than in the genetic disorders.
21. Xanthurenate
Xanthurenate is formed from an amino acid that comes from protein in your diet. High levels can indicate an insufficiency of B6, a vitamin critical for all protein metabolism. Use of medications (e.g., oral contraceptives, anti-hypertensives, and bronchodilators) and exposure to tobacco smoke, pesticides, and other agricultural products can all contribute to insufficiency of vitamin B6. Problems with balance, fatigue, and mental/emotional stability (such as PMS and ADHD) are frequently found in patients with inadequate vitamin B6. Research has shown that symptoms of autism can be ameliorated with vitamin B6 supplementation. Additionally, xanthurenate can prevent insulin from performing its vital role in blood sugar regulation, which can contribute to diabetes.
4. Full panel organic acids.
1 Citramalic (Citramal) 2 5-hydroxymethyl-2-furoic acid (5-hydrox) 3 3-oxoglutaric acid (3-oxoglu) 4 Furan-2,5-dicarboxylic acid (Furan-2,) 5 Furancarbonylglycine (Furancar) 6 Tartaric acid (Tartaric) 7 Arabinose (Arabinos) 8 Carboxycitric acid (Carboxyc) 9 Tricarballytic (Tricarba) 10 2-hydroxyphenylacetic acid (2-hydrox) 11 4-hydroxyphenylacetic acid (4-hydrox) 12 4-Hydroxybenzoic (4hbenzoi) 13 4-Hydroxyhippuric (4hhippuu) 14 Hippuric acid (Hippuric) 15 3-indoleacetic acid (3-indole) 16 HPHPA (Clostridia marker) (HPHPA) 17 4-Cresol (C. difficile) (4-Cresol) 18 DHPPA (Beneficial bacteria) (DHPPA) 19 Glyceric (Glyceric) 20 Glycolic acid (Glycolic) 21 Oxalic acid (Oxalic a) 22 Lactic acid (Lactic a) 23 Pyruvic acid (Pyruvic ) 24 2-hydroxybutyric (2-hydrox) 25 Succinic acid (Succinic) 26 Fumaric acid (Fumaric ) 27 Malic acid (Malic) 28 2-oxo-glutaric acid (2-oxo-gl) 29 Aconitic acid (Aconitic) 30 Citric acid (Citric a) 31 HVA (Homovanilic) (Dopamine) (HVA) 32 VMA (Vanillylmandelic) ((nor)-epinephrine) (VMA Urin) 33 HVA/VMA Ratio (HVA/VMA) 34 5-hydroxyindoleacetic acid (serotonin) (5-hydrox) 35 Quinolinic acid (Quinolin) 36 Kynurenic acid (Kynureni) 37 Uracil (Uracil) 38 Thymine (Thymine) 39 3-hydroxybutyric acid (3-hydrox) 40 Acetoacetic acid (Acetoace) 41 4-hydroxybutyric acid (4-hydrox) 42 Ethylmalonic acid (Ethylmal) 43 Methylsuccinic acid (Methylsu) 44 Adipic acid (Adipic a) 45 Suberic acid (Suberic ) 46 Sebacic (Sebacic) 47 Methylmalonic acid (GPLacids) 48 Pyridoxic (Pyridoc) 49 Pantothenic (Pantothe) 50 Glutaric acid (Glutaric) 51 Succinylacetone 52 3-hydroxy-3-methylglutaric acid (3-hydrox) 53 N-Acetylcysteine (AcetylCy) 54 Methylcitric (Methcitr) 55 Pyroglutamic acid (Pyroglut) 56 Orotic acid (Orotic a) 57 Hydroxyhippuric acid (Hydroxyh) 58 2-hydroxyisovaleric acid (2-hydrox) 59 2-oxoisovaaleric acid (2-oxoiso) 60 3-methyl-2-oxovaleric acid (3-methyl) 61 Hydroxyisovaleric acid (Hydroxyi) 62 2-oxoisocaproic acid (2-oxoiso) 63 2-oxo-4-methylbutyric acid (2-oxo-4-) 64 Mandelic acid (Mandelic) 65 Phenyllactic acid (Phenylla) 66 Phenylpyruvic acid (Phenylpy) 67 Homogentisic acid (Homogent) 68 4-hydroxyphenyllactic (4-hydrox) 69 N-acetyl-aspartic acid (N-acetyl) 70 Malonic acid (Malonic ) 71 Methylglutaric acid (Methylgl) 72 3-Hydroxyglutaric (Hydroxyg) 73 3-Methylglutaconic (3-Methyl) 74 3,4-Dihydroxyhydrocinnamic acid (Cinnamic) 75 Benzoic acid (Benzoic) 76 Isocitric acid (IsocitrU) 77 Figlu (FigMSOch) 78 Picolinic acid (Picoline) 79 Pimelic acid (PimelinU) 80 90. Xanthurenic acid (90.XantU) 81 2-Methylhippuric acid (M-HippuU) 82 3-methylhippuurzuur + 4-methylhippuurzuur (hippuric) 83 Hexanoyl glycine (HexGly U) 84 Phenylglyoxylic acid (Benzoylformic acid) (FenylglU) 85 Glucaric Acid (Saccharic acid) (GlucariU) 86 3-Hydroxypropanoic acid (Propanoi) 87 3-Methylcrotonyl Glycine (Crotonyl) 88 2-Hydroxyglutaric acid (HydroGlu) 89 Alpha-Ketobutyric acid (KetoButy) 90 Mevalonolactone (mevaloU) 91 N-(3-Phenylpropionyl)glycine (N3Pheylp) 92 Propionyl Glycine (2-propanamidoacetic acid) (PropiGly) 93 Suberyl glycine (SuberylU) 94 Tiglyl glycine (Tiglyl-g) New Paragraph
