Phenol red

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Phenol red
Phenol-red-zwitterionic-form-2D-skeletal.png
Phenol-red-zwitterionic-form-3D-balls.png
Identifiers
3D model (JSmol)
ChEBI
ChEMBL
ChemSpider
ECHA InfoCard 100.005.100 OOjs UI icon edit-ltr-progressive.svg
PubChem CID
UNII
  • InChI=1S/C19H14O5S/c20-15-9-5-13(6-10-15)19(14-7-11-16(21)12-8-14)17-3-1-2-4-18(17)25(22,23)24-19/h1-12,20-21H Yes check.svgY
    Key: BELBBZDIHDAJOR-UHFFFAOYSA-N Yes check.svgY
  • O=S2(=O)OC(c1ccccc12)(c3ccc(O)cc3)c4ccc(O)cc4
Properties
C19H14O5S
Molar mass 354.38 g·mol−1
Pharmacology
V04CH03 ( WHO )
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
Yes check.svgY  verify  (what is  Yes check.svgYX mark.svgN ?)

Phenol red (also known as phenolsulfonphthalein or PSP) is a pH indicator frequently used in cell biology laboratories.

Chemical structure and properties

Phenol red exists as a red crystal that is stable in air. Its solubility is 0.77 grams per liter (g/L) in water and 2.9 g/L in ethanol. [1] It is a weak acid with pKa = 8.00 at 20 °C (68 °F).

A solution of phenol red is used as a pH indicator, often in cell culture. Its color exhibits a gradual transition from yellow (λmax = 443 nm [2] ) to red (λmax = 570 nm [3] ) over the pH range 6.8 to 8.2. Above pH 8.2, phenol red turns a bright pink (fuchsia) color. [4] [5]

Phenol red(pH indicator)
below pH 6.8above pH 8.2
6.88.2

In crystalline form, and in solution under very acidic conditions (low pH), the compound exists as a zwitterion as in the structure shown above, with the sulfate group negatively charged, and the ketone group carrying an additional proton. This form is sometimes symbolically written as H+
2
PS
and is orange-red. If the pH is increased (pKa = 1.2), the proton from the ketone group is lost, resulting in the yellow, negatively charged ion denoted as HPS. At still higher pH (pKa = 7.7), the phenol's hydroxy group loses its proton, resulting in the red ion denoted as PS2−. [6]

In several sources, the structure of phenol red is shown with the sulfur atom being part of a cyclic group, similar to the structure of phenolphthalein. [1] [7] However, this cyclic structure could not be confirmed by X-ray crystallography. [8]

Several indicators share a similar structure to phenol red, including bromothymol blue, thymol blue, bromocresol purple, thymolphthalein, and phenolphthalein. (A table of other common chemical indicators is available in the article on pH indicators.)

Uses

Phenolsulfonphthalein test

Phenol red was used by Leonard Rowntree and John Geraghty in the phenolsulfonphthalein test to estimate the overall blood flow through the kidney in 1911. [9] It was the first test of kidney function and was used for almost a century but is now obsolete.

The test is based on the fact that phenol red is excreted almost entirely in the urine. Phenol red solution is administered intravenously; the urine produced is collected. By measuring the amount of phenol red excreted colorimetrically, kidney function can be determined. [10]

Indicator for cell cultures

Phenol red, 40 mM: colors in cell culture medium at a pH range from 6.0 to 8.0. Phenol red pH 6,0 - 8,0.jpg
Phenol red, 40 μM: colors in cell culture medium at a pH range from 6.0 to 8.0.

Most living tissues prosper at a near-neutral pH—that is, a pH close to 7. The pH of blood ranges from 7.35 to 7.45, for instance. When cells are grown in tissue culture, the medium in which they grow is held close to this physiological pH. A small amount of phenol red added to this growth medium will have a pink-red color under normal conditions. Typically, 15 mg/L are used for cell culture.

In the event of problems, waste products produced by dying cells or overgrowth of contaminants will cause a change in pH, leading to a change in indicator color. For example, a culture of relatively slowly dividing mammalian cells can be quickly overgrown by bacterial contamination. This usually results in an acidification of the medium, turning it yellow. Many biologists find this a convenient way to rapidly check on the health of tissue cultures. In addition, the waste products produced by the mammalian cells themselves will slowly decrease the pH, gradually turning the solution orange and then yellow. This color change is an indication that even in the absence of contamination, the medium needs to be replaced (generally, this should be done before the medium has turned completely orange).

Since the color of phenol red can interfere with some spectrophotometric and fluorescent assays, many types of tissue culture media are also available without phenol red.

Estrogen mimic

A commercial test kit for swimming pools with phenol red and an orthotolidine indicator solution Swimming Pool Test Kit.jpg
A commercial test kit for swimming pools with phenol red and an orthotolidine indicator solution

Phenol red is a weak estrogen mimic, and in cell cultures can enhance the growth of cells that express the estrogen receptor. [11] It has been used to induce ovarian epithelial cells from post-menopausal women to differentiate into cells with properties of oocytes (eggs), with potential implications for both fertility treatment and stem cell research. [12]

Use in swimming pool test kits

Phenol red, sometimes labelled with a different name, such as "Guardex Solution #2", is used as a pH indicator in home swimming pool test kits. [13]

Chlorine can result in the bleaching of the dye in the absence of thiosulfate to inhibit the oxidizing chlorine. High levels of bromine can convert phenol red to bromophenol red (dibromophenolsulfonephthalein, whose lowered pKa results in an indicator with a range shifted in the acidic direction – water at pH 6.8 will appear to test at 7.5). Even higher levels of bromine (>20 ppm) can result in the secondary conversion of bromophenol red to bromophenol blue with an even lower pKa, erroneously giving the impression that the water has an extremely high pH despite being dangerously low. [14]

Related Research Articles

<span class="mw-page-title-main">Titration</span> Laboratory method for determining the concentration of an analyte

Titration is a common laboratory method of quantitative chemical analysis to determine the concentration of an identified analyte. A reagent, termed the titrant or titrator, is prepared as a standard solution of known concentration and volume. The titrant reacts with a solution of analyte to determine the analyte's concentration. The volume of titrant that reacted with the analyte is termed the titration volume.

A pH indicator is a halochromic chemical compound added in small amounts to a solution so the pH (acidity or basicity) of the solution can be determined visually or spectroscopically by changes in absorption and/or emission properties. Hence, a pH indicator is a chemical detector for hydronium ions (H3O+) or hydrogen ions (H+) in the Arrhenius model. Normally, the indicator causes the color of the solution to change depending on the pH. Indicators can also show change in other physical properties; for example, olfactory indicators show change in their odor. The pH value of a neutral solution is 7.0 at 25°C (standard laboratory conditions). Solutions with a pH value below 7.0 are considered acidic and solutions with pH value above 7.0 are basic. Since most naturally occurring organic compounds are weak electrolytes, such as carboxylic acids and amines, pH indicators find many applications in biology and analytical chemistry. Moreover, pH indicators form one of the three main types of indicator compounds used in chemical analysis. For the quantitative analysis of metal cations, the use of complexometric indicators is preferred, whereas the third compound class, the redox indicators, are used in redox titrations (titrations involving one or more redox reactions as the basis of chemical analysis).

<span class="mw-page-title-main">Phenolphthalein</span> pH indicator turning to colorless – in basic solution

Phenolphthalein ( feh-NOL(F)-thə-leen) is a chemical compound with the formula C20H14O4 and is often written as "HIn", "HPh", "phph" or simply "Ph" in shorthand notation. Phenolphthalein is often used as an indicator in acid–base titrations. For this application, it turns colorless in acidic solutions and pink in basic solutions. It belongs to the class of dyes known as phthalein dyes.

<span class="mw-page-title-main">Bromothymol blue</span> pH indicator

Bromothymol blue is a pH indicator. It is mostly used in applications that require measuring substances that would have a relatively neutral pH. A common use is for measuring the presence of carbonic acid in a liquid. It is typically sold in solid form as the sodium salt of the acid indicator.

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

Bromophenol blue, albutest is used as a pH indicator, an electrophoretic color marker, and a dye. It can be prepared by slowly adding excess bromine to a hot solution of phenolsulfonphthalein in glacial acetic acid.

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

Methyl orange is a pH indicator frequently used in titration because of its clear and distinct color variance at different pH values. Methyl orange shows red color in acidic medium and yellow color in basic medium. Because it changes color at the pKa of a mid strength acid, it is usually used in titration of strong acids in weak bases that reach the equivalence point at a pH of 3.1-4.4. Unlike a universal indicator, methyl orange does not have a full spectrum of color change, but it has a sharp end point. In a solution becoming less acidic, methyl orange changes from red to orange and, finally, to yellow—with the reverse process occurring in a solution of increasing acidity.

<span class="mw-page-title-main">Universal indicator</span> Indicator that works over a wide range of pH

A universal indicator is a pH indicator made of a solution of several compounds that exhibit various smooth colour changes over a wide range pH values to indicate the acidity or alkalinity of solutions. A universal indicator can be in paper form or present in a form of a solution.

<span class="mw-page-title-main">Methyl red</span> Chemical indicator that turns red in acidic solutions

Methyl red (2-(N,N-dimethyl-4-aminophenyl) azobenzenecarboxylic acid), also called C.I. Acid Red 2, is an indicator dye that turns red in acidic solutions. It is an azo dye, and is a dark red crystalline powder. Methyl red is a pH indicator; it is red in pH under 4.4, yellow in pH over 6.2, and orange in between, with a pKa of 5.1. Murexide and methyl red are investigated as promising enhancers of sonochemical destruction of chlorinated hydrocarbon pollutants. Methyl red is classed by the IARC in group 3 - unclassified as to carcinogenic potential in humans.

<span class="mw-page-title-main">Kastle–Meyer test</span> Blood test utilizing phenolphthalein

The Kastle–Meyer test is a presumptive blood test, first described in 1903, in which the chemical indicator phenolphthalein is used to detect the possible presence of hemoglobin. It relies on the peroxidase-like activity of hemoglobin in blood to catalyze the oxidation of phenolphthalin into phenolphthalein, which is visible as a bright pink color. The Kastle–Meyer test is a form of catalytic blood test, one of the two main classes of forensic tests commonly employed by crime labs in the chemical identification of blood. The other class of tests used for this purpose are microcrystal tests, such as the Teichmann crystal test and the Takayama crystal test.

<span class="mw-page-title-main">Rapid urease test</span> Test for Heliobacter pylori infection

Rapid urease test, also known as the CLO test, is a rapid diagnostic test for diagnosis of Helicobacter pylori. The basis of the test is the ability of H. pylori to secrete the urease enzyme, which catalyzes the conversion of urea to ammonia and carbon dioxide.

<span class="mw-page-title-main">Bromocresol green</span> Chemical dye and pH indicator

Bromocresol green (BCG) is a dye of the triphenylmethane family. It belongs to a class of dyes called sulfonephthaleins. It is used as a pH indicator in applications such as growth mediums for microorganisms and titrations. In clinical practise, it is commonly used as a diagnostic technique. The most common use of bromocresol green is to measure serum albumin concentration within mammalian blood samples in possible cases of kidney failure and liver disease. In chemistry, bromocresol green is used in Thin-layer chromatography staining solutions to visualize acidic compounds.

<span class="mw-page-title-main">XLD agar</span> Selective culture medium

Xylose Lysine Deoxycholate agar is a selective growth medium used in the isolation of Salmonella and Shigella species from clinical samples and from food. The agar was developed by Welton Taylor in 1965. It has a pH of approximately 7.4, leaving it with a bright pink or red appearance due to the indicator phenol red. Sugar fermentation lowers the pH and the phenol red indicator registers this by changing to yellow. Most gut bacteria, including Salmonella, can ferment the sugar xylose to produce acid; Shigella colonies cannot do this and therefore remain red. After exhausting the xylose supply Salmonella colonies will decarboxylate lysine, increasing the pH once again to alkaline and mimicking the red Shigella colonies. Salmonellae metabolise thiosulfate to produce hydrogen sulfide, which leads to the formation of colonies with black centers and allows them to be differentiated from the similarly coloured Shigella colonies.

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

Bromocresol purple (BCP) or 5′,5″-dibromo-o-cresolsulfophthalein, is a dye of the triphenylmethane family and a pH indicator. It is colored yellow below pH 5.2, and violet above pH 6.8. In its cyclic sulfonate ester form, it has a pKa value of 6.3, and is usually prepared as a 0.04% aqueous solution.

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

Chlorophenol red is an indicator dye that changes color from yellow to violet in the pH range 5.4 to 6.8. The pH of a substance is determined by taking the negative logarithm of the Hydronium ion concentration and the indictor changes color due to the dissociation of H+ ions. The lambda max is at 572 nm.

<span class="mw-page-title-main">Dermatophyte test medium</span>

Dermatophyte test medium (DTM) is a specialized agar used in medical mycology. It is based on Sabouraud's dextrose agar with added cycloheximide to inhibit saprotrophic growth, antibiotic to inhibit bacterial growth, and phenol red a pH indicator. The pH indicator is useful in distinguishing a dermatophyte fungus, which utilizes nitrogenous material for preferred metabolism, producing alkaline by-products, imparting a red color change to the medium. Typical saprotrophic fungi utilize carbohydrates in the medium producing acidic by-products and no red color change.

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

Azo violet (Magneson I; p-nitrobenzeneazoresorcinol) is an azo compound with the chemical formula C12H9N3O4. It is used commercially as a violet dye and experimentally as a pH indicator, appearing yellow below pH 11, and violet above pH 13. It also turns deep blue in the presence of magnesium salt in a slightly alkaline, or basic, environment. Azo violet may also be used to test for the presence of ammonium ions. The color of ammonium chloride or ammonium hydroxide solution will vary depending upon the concentration of azo violet used. Magneson I is used to test Be also; it produces an orange-red lake with Be(II) in alkaline medium.

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

Bromopyrogallol red is frequently used in analytical chemistry as a reagent for spectrophometric analysis and as an complexometric indicator.

E-SCREEN is a cell proliferation assay based on the enhanced proliferation of human breast cancer cells (MCF-7) in the presence of estrogen active substances. The E-SCREEN test is a tool to easily and rapidly assess estrogenic activity of suspected xenoestrogens. This bioassay measures estrogen-induced increase of the number of human breast cancer cell, which is biologically equivalent to the increase of mitotic activity in tissues of the genital tract. It was originally developed by Soto et al. and was included in the first version of the OECD Conceptual Framework for Testing and Assessment of Endocrine Disrupters published in 2012. However, due to failed validation, it was not included in the updated version of the framework published in 2018.

Diagnostic microbiology is the study of microbial identification. Since the discovery of the germ theory of disease, scientists have been finding ways to harvest specific organisms. Using methods such as differential media or genome sequencing, physicians and scientists can observe novel functions in organisms for more effective and accurate diagnosis of organisms. Methods used in diagnostic microbiology are often used to take advantage of a particular difference in organisms and attain information about what species it can be identified as, which is often through a reference of previous studies. New studies provide information that others can reference so that scientists can attain a basic understanding of the organism they are examining.

<span class="mw-page-title-main">Phthalein dye</span>

Phthalein dyes are a class of dyes mainly used as pH indicators, due to their ability to change colors depending on pH. They are formed by the reaction of phthalic anhydride with various phenols. They are a subclass of triarylmethane dyes.

References

  1. 1 2 Merck Index , 11th ed., 7213 Phenolsulfonphtalein
  2. Saha, U.; Mukherjea, K. K. (2015). "Development of a multifunctional biomimicking L-cysteine based oxovanadium(IV) complex: synthesis, DFT calculations, bromo-peroxidation and nuclease activity". RSC Advances. 5 (114): 94462–94473. doi:10.1039/C5RA19585C.
  3. Mills, A.; Skinner, G. A. (2011). "A novel 'fizziness' indicator". The Analyst. 136 (5): 894–896. Bibcode:2011Ana...136..894M. doi:10.1039/c0an00610f. PMID   21210046.
  4. Merck Index , 13th ed., 7329 Phenolsulfonphthalein
  5. Beilstein 5-19-03-00457
  6. Tamura, Z.; Maeda M. (1997). "Differences between phthaleins and sulfonphthaleins". Yakugaku Zasshi (in Japanese). 117 (10–11): 764–770. doi: 10.1248/yakushi1947.117.10-11_764 . PMID   9414589.
  7. "Phenolsulfonphthalein". PubChem . NIH.
  8. Yamaguchi, K.; Tamura, Z.; Maeda, M. (1997). "Molecular Structure of the Zwitterionic Form of Phenolsulfonphthalein". Analytical Sciences. 13 (3): 521–522. doi: 10.2116/analsci.13.521 .
  9. GERAGHTY, J. T.; ROWNTREE, L. G. (2 September 1911). "The Phenolsulphonephthalein Test for Estimating Renal Function". Journal of the American Medical Association. LVII (10): 811. doi:10.1001/jama.1911.04260090033014.
  10. "Phenolsulfonphthalein Test". Encyclopædia Britannica .
  11. Berthois, Y.; Katzenellenbogen, J. A.; Katzenellenbogen, B. S. (1986). "Phenol red in tissue culture media is a weak estrogen: Implications concerning the study of estrogen-responsive cells in culture". Proceedings of the National Academy of Sciences of the United States of America. 83 (8): 2496–2500. Bibcode:1986PNAS...83.2496B. doi: 10.1073/pnas.83.8.2496 . PMC   323325 . PMID   3458212.
  12. Bukovsky, A.; Svetlikova, M.; Caudle, M. R. (2005). "Oogenesis in Cultures Derived from Adult Human Ovaries" (PDF). Reproductive Biology and Endocrinology. 3 (5): 17. doi: 10.1186/1477-7827-3-17 . PMC   1131924 . PMID   15871747.
  13. Guardex Solution 2 – Phenol Red Material Safety Data Sheet
  14. Effect of Bromine on Phenol Red in pH Tests