Bromothymol blue

Last updated
Bromothymol blue
Bromothymol-blue-2D-skeletal.png
Bromothymol-blue-3D-balls.png
Names
Preferred IUPAC name
3,3-Bis[3-bromo-4-hydroxy-2-methyl-5-(propan-2-yl)phenyl]-2,1λ6-benzoxathiole-1,1(3H)-dione
Identifiers
3D model (JSmol)
ChEBI
ChEMBL
ChemSpider
ECHA InfoCard 100.000.884 OOjs UI icon edit-ltr-progressive.svg
EC Number
  • 200-971-2
PubChem CID
UNII
  • InChI=1S/C27H28Br2O5S/c1-13(2)17-11-20(15(5)23(28)25(17)30)27(19-9-7-8-10-22(19)35(32,33)34-27)21-12-18(14(3)4)26(31)24(29)16(21)6/h7-14,30-31H,1-6H3 Yes check.svgY
    Key: NUHCTOLBWMJMLX-UHFFFAOYSA-N Yes check.svgY
  • InChI=1/C27H28Br2O5S/c1-13(2)17-11-20(15(5)23(28)25(17)30)27(19-9-7-8-10-22(19)35(32,33)34-27)21-12-18(14(3)4)26(31)24(29)16(21)6/h7-14,30-31H,1-6H3
    Key: NUHCTOLBWMJMLX-UHFFFAOYAD
  • Brc1c(O)c(cc(c1C)C3(OS(=O)(=O)c2ccccc23)c4cc(c(O)c(Br)c4C)C(C)C)C(C)C
Properties
C27H28Br2O5S
Molar mass 624.38 g·mol−1
Density 1.25 g/cm3
Melting point 202 °C (396 °F; 475 K)
Sparingly soluble in water [1]
Acidity (pKa)7.0
Hazards
GHS labelling:
GHS-pictogram-exclam.svg
Warning
H302, H315, H319
P264, P270, P280, P301+P312, P302+P352, P305+P351+P338, P321, P330, P332+P313, P337+P313, P362, P501
NFPA 704 (fire diamond)
NFPA 704.svgHealth 2: Intense or continued but not chronic exposure could cause temporary incapacitation or possible residual injury. E.g. chloroformFlammability 1: Must be pre-heated before ignition can occur. Flash point over 93 °C (200 °F). E.g. canola oilInstability 0: Normally stable, even under fire exposure conditions, and is not reactive with water. E.g. liquid nitrogenSpecial hazards (white): no code
2
1
0
Safety data sheet (SDS) ThermoFisher Scientific
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
X mark.svgN  verify  (what is  Yes check.svgYX mark.svgN ?)

Bromothymol blue (also known as bromothymol sulfone phthalein and BTB) is a pH indicator. It is mostly used in applications that require measuring substances that would have a relatively neutral pH (near 7). 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.

Contents

Structure and properties

Bromothymol blue(pH indicator)
below pH 6.0above pH 7.6
6.07.6

Bromothymol blue acts as a weak acid in a solution. It can thus be in protonated or deprotonated form, appearing yellow or blue, respectively. It is bright aquamarine by itself, and greenish-blue in a neutral solution. The deprotonation of the neutral form results in a highly conjugated structure, accounting for the difference in color. An intermediate of the deprotonation mechanism is responsible for the greenish color in neutral solution. [2]

The protonated form of bromothymol blue has its peak absorption at 427 nm thus transmitting yellow light in acidic solutions, and the deprotonated form has its peak absorption at 602 nm thus transmitting blue light in more basic solutions. [3] Highly acidic Bromothymol blue is magenta in color.

The general carbon skeleton of bromothymol blue is common to many indicators including chlorophenol red, thymol blue, and bromocresol green. [2]

The presence of one moderate electron-withdrawing group (bromine atom) and two moderate donating groups (alkyl substituents) are responsible for bromothymol blue's active indication range from a pH of 6.0 to 7.6. While the conjugation is responsible for the length and nature of the color change range, these substituent groups are ultimately responsible for the indicator's active range. [2]

Yellow acidic form (left) and blue basic form (right) of bromothymol blue. Bromothymol blue deprotonation.svg
Yellow acidic form (left) and blue basic form (right) of bromothymol blue.

Bromothymol blue is sparingly soluble in oil, but soluble in water, ether, and aqueous solutions of alkalis. It is less soluble in nonpolar solvents such as benzene, toluene, and xylene, and practically insoluble in petroleum ether. [5]

Synthesis and preparation

Bromothymol blue is synthesized by addition of elemental bromine to thymol blue in a solution in glacial acetic acid. [6]

To prepare a solution for use as pH indicator, dissolve 0.10 g in 8.0 cm3 N/50 (a.k.a. 0.02 Normal) NaOH and dilute with water to 250 cm3. To prepare a solution for use as indicator in volumetric work, dissolve 0.1 g in 100 cm3 of 50% (v/v) ethanol. [5]

Uses

Different colors of bromothymol blue at marked pH conditions. Bromothymol blue colors at different pH.png
Different colors of bromothymol blue at marked pH conditions.

Bromothymol blue may be used for observing photosynthetic activities, or as a respiratory indicator (turns yellow as CO2 is added). [7] [8] A common demonstration of BTB's pH indicator properties involves exhaling through a tube into a neutral solution of BTB. As CO2 is absorbed from the breath into the solution, forming carbonic acid, the solution changes color from green to yellow. Thus, BTB is commonly used in science classes to demonstrate that the more that muscles are used, the greater the CO2 output.

Bromothymol blue has been used in conjunction with phenol red to monitor the fungal asparaginase enzyme activity with phenol red turning pink and bromothymol blue turning blue indicating an increase in pH and therefore enzyme activity. [9] However, a recent study suggests that methyl red is more useful in determining activity due to the bright yellow ring formed in the zone of enzyme activity. [10]

It may also be used in the laboratory as a biological slide stain. At this point, the bromothymol is already blue, and a few drops of BTB are used on a water slide. The specimen is mixed with blue BTB solution and fixed to a slide by a cover slip. It is sometimes used to define cell walls or nuclei under the microscope.

Bromothymol is used in obstetrics for detecting premature rupture of membranes. [11] Amniotic fluid typically has a pH > 7.2, bromothymol will therefore turn blue when brought in contact with fluid leaking from the amnion. As vaginal pH normally is acidic, the blue color indicates the presence of amniotic fluid. The test may be false-positive in the presence of other alkaline substances such as blood or semen, or in the presence of bacterial vaginosis.

See also

Related Research Articles

<span class="mw-page-title-main">Carboxylic acid</span> Organic compound containing a –C(=O)OH group

In organic chemistry, a carboxylic acid is an organic acid that contains a carboxyl group attached to an R-group. The general formula of a carboxylic acid is often written as R−COOH or R−CO2H, sometimes as R−C(O)OH with R referring to the alkyl, alkenyl, aryl, or other group. Carboxylic acids occur widely. Important examples include the amino acids and fatty acids. Deprotonation of a carboxylic acid gives a carboxylate anion.

<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.

<span class="mw-page-title-main">Base (chemistry)</span> Type of chemical substance

In chemistry, there are three definitions in common use of the word "base": Arrhenius bases, Brønsted bases, and Lewis bases. All definitions agree that bases are substances that react with acids, as originally proposed by G.-F. Rouelle in the mid-18th century.

<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">Methyl violet</span> Chemical compound

Methyl violet is a family of organic compounds that are mainly used as dyes. Depending on the number of attached methyl groups, the color of the dye can be altered. Its main use is as a purple dye for textiles and to give deep violet colors in paint and ink. It is also used as a hydration indicator for silica gel. Methyl violet 10B is also known as crystal violet and has medical uses.

<span class="mw-page-title-main">Fluorescein</span> Synthetic organic compound used as dye and fluorescent tracer

Fluorescein is an organic compound and dye based on the xanthene tricyclic structural motif, formally belonging to triarylmethine dyes family. It is available as a dark orange/red powder slightly soluble in water and alcohol. It is widely used as a fluorescent tracer for many applications.

<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.

Classical qualitative inorganic analysis is a method of analytical chemistry which seeks to find the elemental composition of inorganic compounds. It is mainly focused on detecting ions in an aqueous solution, therefore materials in other forms may need to be brought to this state before using standard methods. The solution is then treated with various reagents to test for reactions characteristic of certain ions, which may cause color change, precipitation and other visible changes.

<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">Phenol red</span> Chemical compound

Phenol red is a pH indicator frequently used in cell biology laboratories.

<span class="mw-page-title-main">Litmus</span> Substance to test chemical acidity

Litmus is a water-soluble mixture of different dyes extracted from lichens. It is often absorbed onto filter paper to produce one of the oldest forms of pH indicator, used to test materials for acidity. In an acidic medium, blue litmus paper turns red, while in a basic or alkaline medium, red litmus paper turns blue. In short, it is a dye and indicator which is used to place substances on a pH scale.

<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">Thymol blue</span> Chemical compound

Thymol blue (thymolsulfonephthalein) is a brownish-green or reddish-brown crystalline powder that is used as a pH indicator. It is insoluble in water but soluble in alcohol and dilute alkali solutions.

<span class="mw-page-title-main">4-Nitrophenol</span> Chemical compound

4-Nitrophenol is a phenolic compound that has a nitro group at the opposite position of the hydroxyl group on the benzene ring.

<span class="mw-page-title-main">IMViC</span> Microbiological and biochemical method for identification

The IMViC tests are a group of individual tests used in microbiology lab testing to identify an organism in the coliform group. A coliform is a gram negative, aerobic, or facultative anaerobic rod, which produces gas from lactose within 48 hours. The presence of some coliforms indicate fecal contamination.

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

Alcian blue is any member of a family of polyvalent basic dyes, of which the Alcian blue 8G has been historically the most common and the most reliable member. It is used to stain acidic polysaccharides such as glycosaminoglycans in cartilages and other body structures, some types of mucopolysaccharides, sialylated glycocalyx of cells etc. For many of these targets it is one of the most widely used cationic dyes for both light and electron microscopy. Use of alcian blue has historically been a popular staining method in histology especially for light microscopy in paraffin embedded sections and in semithin resin sections. The tissue parts that specifically stain by this dye become blue to bluish-green after staining and are called "Alcianophilic". Alcian blue staining can be combined with H&E staining, PAS staining and van Gieson staining methods. Alcian blue can be used to quantitate acidic glycans both in microspectrophotometric quantitation in solution or for staining glycoproteins in polyacrylamide gels or on western blots. Biochemists had used it to assay acid polysaccharides in urine since the 1960s for diagnosis of diseases like mucopolysaccharidosis but from 1970's, partly due to lack of availability of Alcian and partly due to length and tediousness of the procedure, alternative methods had to be developed e.g. Dimethyl methylene blue method.

<span class="mw-page-title-main">Thiosulfate–citrate–bile salts–sucrose agar</span>

Thiosulfate–citrate–bile salts–sucrose agar, or TCBS agar, is a type of selective agar culture plate that is used in microbiology laboratories to isolate Vibrio species. TCBS agar is highly selective for the isolation of V. cholerae and V. parahaemolyticus as well as other Vibrio species. Apart from TCBS agar, other rapid testing dipsticks like immunochromatographic dipstick is also used in endemic areas such as Asia, Africa and Latin America. Though, TCBS agar study is required for confirmation. This becomes immensely important in cases of gastroenteritis caused by campylobacter species, whose symptoms mimic that of cholera. Since no yellow bacterial growth is observed in case of campylobacter species on TCBS agar, chances of incorrect diagnosis can be rectified. TCBS agar contains high concentrations of sodium thiosulfate and sodium citrate to inhibit the growth of Enterobacteriaceae. Inhibition of gram-positive bacteria is achieved by the incorporation of ox gall, which is a naturally occurring substance containing a mixture of bile salts and sodium cholate, a pure bile salt. Sodium thiosulfate also serves as a sulfur source and its presence, in combination with ferric citrate, allows for the easy detection of hydrogen sulfide production. Saccharose (sucrose) is included as a fermentable carbohydrate for metabolism by Vibrio species. The alkaline pH of the medium enhances the recovery of V. cholerae and inhibits the growth of others. Thymol blue and bromothymol blue are included as indicators of pH changes.

<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.

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

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References

  1. "Archived copy". Archived from the original on 2015-11-26. Retrieved 2015-12-11.{{cite web}}: CS1 maint: archived copy as title (link)
  2. 1 2 3 De Meyer, Thierry (March 2014). "Substituent effects on absorption spectra of pH indicators: An experimental and computational study of sulfonphthaleine dyes". Dyes and Pigments. 102: 241–250. doi:10.1016/j.dyepig.2013.10.048. hdl: 1854/LU-4353650 . Archived from the original on 2020-07-29. Retrieved 2020-03-16.
  3. Nahhal; et al. (18 July 2012). "Thin film optical BTB pH sensors using sol–gel method in presence of surfactants" (PDF). International Nano Letters. 2 (16): 3. Bibcode:2012INL.....2...16E. doi: 10.1186/2228-5326-2-16 . Archived (PDF) from the original on 2014-11-29. Retrieved 18 November 2014.
  4. Klotz, Elsbeth; Doyle, Robert; Gross, Erin; Mattson, Bruce (2011). "The Equilibrium Constant for Bromothymol Blue: A General Chemistry Laboratory Experiment Using Spectroscopy". J. Chem. Educ. 88 (5): 637–639. Bibcode:2011JChEd..88..637K. doi:10.1021/ed1007102 . Retrieved 2023-02-07.
  5. 1 2 O'Neil, Maryadele J (2006). The Merck Index. Merck Research Laboratory. p. 1445. ISBN   978-0-911910-00-1.
  6. "Bromothymol blue" (PDF). Archived (PDF) from the original on 27 November 2020. Retrieved 4 April 2020.
  7. Sabnis R. W. (2007). Handbook of Acid-Base Indicators. CRC Press. ISBN   978-0-8493-8218-5.
  8. Sabnis R. W. (2010). Handbook of Biological Dyes and Stains: Synthesis and Industrial Applications (1st ed.). Wiley. ISBN   978-0-470-40753-0.
  9. "Isolation and screening of L-asparaginase free of glutaminase and urease from fungal sp". researchgate.net. 22 August 2016.
  10. Dhale, Mohan (July 2014). "A comparative rapid and sensitive method to screen l-asparaginase producing fungi". Journal of Microbiological Methods. 102: 66–68. doi:10.1016/j.mimet.2014.04.010. PMID   24794733.
  11. King, Arthur G. (1935-12-01). "The determination of rupture of the membranes". American Journal of Obstetrics & Gynecology. 30 (6): 860–862. doi:10.1016/S0002-9378(35)90429-X. ISSN   0002-9378.