ABTS

Last updated
ABTS
ABTS.png
Names
IUPAC name
2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
Identifiers
3D model (JSmol)
ChemSpider
PubChem CID
  • InChI=1S/C18H18N4O6S4/c1-3-21-13-7-5-11(31(23,24)25)9-15(13)29-17(21)19-20-18-22(4-2)14-8-6-12(32(26,27)28)10-16(14)30-18/h5-10H,3-4H2,1-2H3,(H,23,24,25)(H,26,27,28)/b19-17-,20-18+ Yes check.svgY
    Key: ZTOJFFHGPLIVKC-YAFCTCPESA-N Yes check.svgY
  • InChI=1/C18H18N4O6S4/c1-3-21-13-7-5-11(31(23,24)25)9-15(13)29-17(21)19-20-18-22(4-2)14-8-6-12(32(26,27)28)10-16(14)30-18/h5-10H,3-4H2,1-2H3,(H,23,24,25)(H,26,27,28)/b19-17-,20-18+
    Key: ZTOJFFHGPLIVKC-YAFCTCPEBW
  • CCN1/C(Sc2cc(ccc12)S(O)(=O)=O)=N/N=C/3Sc4cc(ccc4N3CC)S(O)(=O)=O
  • O=S(=O)(O)c1ccc2N(C(\Sc2c1)=N\N=C4\Sc3cc(ccc3N4CC)S(=O)(=O)O)CC
Properties
C18H18N4O6S4
Molar mass 514.60 g·mol−1
Hazards
GHS labelling:
GHS-pictogram-exclam.svg
Warning
H315, H319, H335
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).

In biochemistry, ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) is a chemical compound used to observe the reaction kinetics of specific enzymes. A common use for it is in the enzyme-linked immunosorbent assay (ELISA) to detect the binding of molecules to each other.

It is commonly used as a substrate with hydrogen peroxide for a peroxidase enzyme (such as horseradish peroxidase) or alone with blue multicopper oxidase enzymes (such as laccase or bilirubin oxidase). Its use allows the reaction kinetics of peroxidases themselves to be followed. In this way it also can be used to indirectly follow the reaction kinetics of any hydrogen peroxide-producing enzyme, or to simply quantify the amount of hydrogen peroxide in a sample.

The formal reduction potentials for ABTS are high enough for it to act as an electron donor for the reduction of oxo species such as molecular oxygen and hydrogen peroxide, particularly at the less-extreme pH values encountered in biological catalysis. Under these conditions, the sulfonate groups are fully deprotonated and the mediator exists as a dianion.

ABTS· + e → ABTS2 = 0.67 V vs SHE
ABTS + e → ABTS· = 1.08 V vs SHE [1]

This compound is chosen because the enzyme facilitates the reaction with hydrogen peroxide, turning it into a green and soluble end-product. Its new absorbance maximum of 420 nm light (ε = 3.6 × 104 M1 cm1) [2] can easily be followed with a spectrophotometer, a common laboratory instrument. It is sometimes used as part of a glucose estimating reagent when finding glucose concentrations of solutions such as blood serum.

ABTS is also frequently used by the food industry and agricultural researchers to measure the antioxidant capacities of foods. [3] In this assay, ABTS is converted to its radical cation by addition of sodium persulfate. This radical cation is blue in color and absorbs light at 415, 645, 734 and 815 nm. [4] The ABTS radical cation is reactive towards most antioxidants including phenolics, thiols and Vitamin C. [5] During this reaction, the blue ABTS radical cation is converted back to its colorless neutral form. The reaction may be monitored spectrophotometrically. This assay is often referred to as the Trolox equivalent antioxidant capacity (TEAC) assay. The reactivity of the various antioxidants tested are compared to that of Trolox, which is a water-soluble analog of vitamin E. [6]

Applications for functional food analysis

Based on the special chemical properties of formed free radicals, ABTS assay has been used to determine the antioxidant capacity of food products. For example, polyphenol compounds, which widely exist in fruit, can quench free radicals inside human body, thus prevent oxidative damage by free radicals. The antioxidant potency of plant extract or food product has been measured by ABTS assay. One example with detailed method is the antioxidant activity analysis of Hibiscus products. [7]

Related Research Articles

Antioxidants are compounds that inhibit oxidation, a chemical reaction that can produce free radicals. Autoxidation leads to degradation of organic compounds, including living matter. Antioxidants are frequently added to industrial products, such as polymers, fuels, and lubricants, to extend their usable lifetimes. Foods are also treated with antioxidants to forestall spoilage, in particular the rancidification of oils and fats. In cells, antioxidants such as glutathione, mycothiol, or bacillithiol, and enzyme systems like superoxide dismutase, can prevent damage from oxidative stress.

<span class="mw-page-title-main">Catalase</span> Enzyme decomposing hydrogen peroxide

Catalase is a common enzyme found in nearly all living organisms exposed to oxygen which catalyzes the decomposition of hydrogen peroxide to water and oxygen. It is a very important enzyme in protecting the cell from oxidative damage by reactive oxygen species (ROS). Catalase has one of the highest turnover numbers of all enzymes; one catalase molecule can convert millions of hydrogen peroxide molecules to water and oxygen each second.

<span class="mw-page-title-main">Cytochrome c peroxidase</span>

Cytochrome c peroxidase, or CCP, is a water-soluble heme-containing enzyme of the peroxidase family that takes reducing equivalents from cytochrome c and reduces hydrogen peroxide to water:

<span class="mw-page-title-main">Polyphenol</span> Class of chemical compounds

Polyphenols are a large family of naturally occurring phenols. They are abundant in plants and structurally diverse. Polyphenols include phenolic acids, flavonoids, tannic acid, and ellagitannin, some of which have been used historically as dyes and for tanning garments.

Lipid peroxidation, or lipid oxidation, is a complex chemical process that leads to oxidative degradation of lipids, resulting in the formation of peroxide and hydroperoxide derivatives. It occurs when free radicals, specifically reactive oxygen species (ROS), interact with lipids within cell membranes, typically polyunsaturated fatty acids (PUFAs) as they have carbon–carbon double bonds. This reaction leads to the formation of lipid radicals, collectively referred to as lipid peroxides or lipid oxidation products (LOPs), which in turn react with other oxidizing agents, leading to a chain reaction that results in oxidative stress and cell damage.

<span class="mw-page-title-main">Oxidative stress</span> Free radical toxicity

Oxidative stress reflects an imbalance between the systemic manifestation of reactive oxygen species and a biological system's ability to readily detoxify the reactive intermediates or to repair the resulting damage. Disturbances in the normal redox state of cells can cause toxic effects through the production of peroxides and free radicals that damage all components of the cell, including proteins, lipids, and DNA. Oxidative stress from oxidative metabolism causes base damage, as well as strand breaks in DNA. Base damage is mostly indirect and caused by the reactive oxygen species generated, e.g., O
2
, OH and H2O2. Further, some reactive oxidative species act as cellular messengers in redox signaling. Thus, oxidative stress can cause disruptions in normal mechanisms of cellular signaling.

Oxygen radical absorbance capacity (ORAC) was a method of measuring antioxidant capacities in biological samples in vitro. Because no physiological proof in vivo existed in support of the free-radical theory or that ORAC provided information relevant to biological antioxidant potential, it was withdrawn in 2012.

<span class="mw-page-title-main">Ascorbate peroxidase</span> Enzyme

Ascorbate peroxidase (or L-ascorbate peroxidase, APX or APEX) (EC 1.11.1.11) is an enzyme that catalyzes the chemical reaction

<span class="mw-page-title-main">Horseradish peroxidase</span> Chemical compound and enzyme

The enzyme horseradish peroxidase (HRP), found in the roots of horseradish, is used extensively in biochemistry applications. It is a metalloenzyme with many isoforms, of which the most studied type is C. It catalyzes the oxidation of various organic substrates by hydrogen peroxide.

<span class="mw-page-title-main">Folin–Ciocalteu reagent</span> Chemical mixture

The Folin–Ciocâlteu reagent (FCR) or Folin's phenol reagent or Folin–Denis reagent, is a mixture of phosphomolybdate and phosphotungstate used for the colorimetric in vitro assay of phenolic and polyphenolic antioxidants, also called the gallic acid equivalence method (GAE). It is named after Otto Folin, Vintilă Ciocâlteu, and Willey Glover Denis. The Folin-Denis reagent is prepared by mixing sodium tungstate and phosphomolybdic acid in phosphoric acid. The Folin–Ciocalteu reagent is just a modification of the Folin-Denis reagent. The modification consisted of the addition of lithium sulfate and bromine to the phosphotungstic-phosphomolybdic reagent.

In enzymology, a lignin peroxidase (EC 1.11.1.14) is an enzyme that catalyzes the chemical reaction

In enzymology, a manganese peroxidase (EC 1.11.1.13) is an enzyme that catalyzes the chemical reaction

<span class="mw-page-title-main">NADH peroxidase</span>

In enzymology, a NADH peroxidase (EC 1.11.1.1) is an enzyme that catalyzes the chemical reaction

<span class="mw-page-title-main">Indicaxanthin</span> Chemical compound

Indicaxanthin is a type of betaxanthin, a plant pigment present in beets, in Mirabilis jalapa flowers, in cacti such as prickly pears or the red dragonfruit. It is a powerful antioxidant.

<span class="mw-page-title-main">Trolox</span> Chemical compound

Trolox is a water-soluble analog of vitamin E sold by Hoffman-LaRoche. It is an antioxidant like vitamin E and it is used in biological or biochemical applications to reduce oxidative stress or damage.

The Trolox equivalent antioxidant capacity (TEAC) assay measures the antioxidant capacity of a given substance, as compared to the standard, Trolox. Most commonly, antioxidant capacity is measured using the ABTS Decolorization Assay. Other antioxidant capacity assays which use Trolox as a standard include the diphenylpicrylhydrazyl (DPPH), oxygen radical absorbance capacity (ORAC) and ferric reducing ability of plasma (FRAP) assays. The TEAC assay is often used to measure the antioxidant capacity of foods, beverages and nutritional supplements.

Colorimetric analysis is a method of determining the concentration of a chemical element or chemical compound in a solution with the aid of a color reagent. It is applicable to both organic compounds and inorganic compounds and may be used with or without an enzymatic stage. The method is widely used in medical laboratories and for industrial purposes, e.g. the analysis of water samples in connection with industrial water treatment.

Haem peroxidases (or heme peroxidases) are haem-containing enzymes that use hydrogen peroxide as the electron acceptor to catalyse a number of oxidative reactions. Most haem peroxidases follow the reaction scheme:

<span class="mw-page-title-main">Grape reaction product</span> Chemical compound

The grape reaction product is a phenolic compound explaining the disappearance of caftaric acid from grape must during processing. It is also found in aged red wines. Its enzymatic production by polyphenol oxidase is important in limiting the browning of musts, especially in white wine production. The product can be recreated in model solutions.

<span class="mw-page-title-main">Fungal extracellular enzyme activity</span> Enzymes produced by fungi and secreted outside their cells

Extracellular enzymes or exoenzymes are synthesized inside the cell and then secreted outside the cell, where their function is to break down complex macromolecules into smaller units to be taken up by the cell for growth and assimilation. These enzymes degrade complex organic matter such as cellulose and hemicellulose into simple sugars that enzyme-producing organisms use as a source of carbon, energy, and nutrients. Grouped as hydrolases, lyases, oxidoreductases and transferases, these extracellular enzymes control soil enzyme activity through efficient degradation of biopolymers.

References

  1. Bourbonnais, Robert; Leech, Dónal; Paice, Michael G. (1998-03-02), "Electrochemical analysis of the interactions of laccase mediators with lignin model compounds", Biochimica et Biophysica Acta (BBA) - General Subjects, 1379 (3): 381–390, doi:10.1016/S0304-4165(97)00117-7, PMID   9545600
  2. Shin, Kwang-Soo; Lee, Yeo-Jin (2000-12-01), "Purification and Characterization of a New Member of the Laccase Family from the White-Rot Basidiomycete Coriolus hirsutus", Archives of Biochemistry and Biophysics, 384 (1): 109–115, doi:10.1006/abbi.2000.2083, PMID   11147821
  3. Huang, Dejian; Ou, Boxin; Prior, Donald L. (2005-02-25), "The Chemistry Behind Antioxidant Capacity Assays", J. Agric. Food Chem., 53 (6): 1841–1856, doi:10.1021/jf030723c, PMID   15769103
  4. Re, Roberta; Pellegrini, Nicoletta; Proteggente, Anna; Pannala, Ananth; Rice-Evans, Catherine (1999-06-02), "Antioxidant activity applying an improved ABTS radical cation decolorization assay", Free Radical Biology and Medicine, 26 (9–10): 1231–1237, doi:10.1016/S0891-5849(98)00315-3, PMID   10381194
  5. Walker, Richard B.; Everette, Jace D. (2009-01-29), "Comparative Reaction Rates of Various Antioxidants with ABTS Radical Cation", J. Agric. Food Chem., 57 (4): 1156–1161, doi:10.1021/jf8026765, PMID   19199590
  6. Barclay, L. R. C.; Locke, S. J.; MacNeil, J. M. (1985), "Autoxidation in micelles - Synergism of Vitamin-C with Lipid-Soluble Vitamin-E and Water-Soluble Trolox", Can. J. Chem., 63 (2): 366–374, doi: 10.1139/v85-062
  7. Zhen, J.; Villani, T. S.; Guo, Y.; Qi, Y.; Chin, K.; Pan, M. H.; Wu, Q. (2016). "Phytochemistry, antioxidant capacity, total phenolic content and anti-inflammatory activity of Hibiscus sabdariffa leaves". Food Chemistry. 190: 673–680. doi:10.1016/j.foodchem.2015.06.006. PMID   26213025.