Identifiers | |
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3D model (JSmol) | |
ChEBI | |
ChEMBL | |
ChemSpider | |
ECHA InfoCard | 100.005.100 |
PubChem CID | |
UNII | |
CompTox Dashboard (EPA) | |
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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). |
Phenol red (also known as phenolsulfonphthalein or PSP) is a pH indicator frequently used in cell biology laboratories.
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 6.8 | above pH 8.2 | |
6.8 | ⇌ | 8.2 |
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+
2PS−
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.)
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]
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.
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]
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]
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.
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.
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.
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.
The enzyme alkaline phosphatase is a phosphatase with the physiological role of dephosphorylating compounds. The enzyme is found across a multitude of organisms, prokaryotes and eukaryotes alike, with the same general function, but in different structural forms suitable to the environment they function in. Alkaline phosphatase is found in the periplasmic space of E. coli bacteria. This enzyme is heat stable and has its maximum activity at high pH. In humans, it is found in many forms depending on its origin within the body – it plays an integral role in metabolism within the liver and development within the skeleton. Due to its widespread prevalence in these areas, its concentration in the bloodstream is used by diagnosticians as a biomarker in helping determine diagnoses such as hepatitis or osteomalacia.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.