6160-80-1

Basic information

• Product Name: 4-Methylumbelliferyl-beta-D-glucuronide
• Synonyms: 4-METHYL-2-OXO-2H-1-BENZOPYRAN-7-YL, BETA-D-GLUCO-PYRANOSIDURONIC ACID;4-METHYLUMBELLIFERYL B-D-GLUCURONIDE;4-METHYLUMBELLIFERYL-BETA-D-GLUCURONIC ACID;4-METHYLUMBELLIFERYL-BETA-D-GLUCURONIDE;MUG;MUGLCU;.beta.-D-Glucopyranosiduronicacid,4-methyl-2-oxo-2H-1-benzopyran-7-yl;4-methylumbelliferylglucuronide
• CAS NO: 6160-80-1
• Molecular Formula: C₁₆H₁₆O₉
• Molecular Weight: 352.29284
• EINECS: 228-186-0
• Product Categories: Substrates;Fluorescent Labels & Indicators;Glucuronides;substrate
• Mol File: 6160-80-1.mol
• Article Data: 11

Chemical Properties

• Melting Point: 102 °C
• Boiling Point: 683.106 °C at 760 mmHg
• Flash Point: 254.961 °C
• Appearance: /
• Density: 1.632 g/cm₃
• Vapor Pressure: 1.35E-19mmHg at 25°C
• Storage Temp.: −20°C
• Solubility: Methanol (Slightly), Water (Slightly)
• PKA: 2.76±0.70(Predicted)
• Water Solubility: slightly soluble
• Sensitive: Moisture Sensitive
• CAS DataBase Reference: 4-Methylumbelliferyl-beta-D-glucuronide(CAS DataBase Reference)
• NIST Chemistry Reference: 4-Methylumbelliferyl-beta-D-glucuronide(6160-80-1)
• EPA Substance Registry System: 4-Methylumbelliferyl-beta-D-glucuronide(6160-80-1)

Safety Data

• Hazard Codes: Xi,Xn
• Statements: 22-40
• Safety Statements: 24/25-36/37
• WGK Germany: 3
• TSCA: Yes
• Hazardous Substances Data: 6160-80-1(Hazardous Substances Data)

Usage

Description

4-Methylumbelliferyl-beta-D-glucuronide is a fluorescent substrate of β-glucuronidase, a hydrolase enzyme that cleaves glucuronic acid from various conjugated compounds. This cleavage results in the release of 4-methylumbelliferone, a blue fluorescent product visible under UV light, which is pH-dependent with excitation maxima of 320 and 360 nm at low (1.97-6.72) and high (7.12-10.3) pH, respectively. It is particularly useful for working with Escherichia coli and plant molecular studies.

Uses

Used in Enzyme Assays:
4-Methylumbelliferyl-beta-D-glucuronide is used as a substrate for β-glucuronidase assays, allowing for the detection and quantification of the enzyme's activity. This application is crucial in various research and diagnostic settings, as it helps in understanding the enzyme's role in biological processes and its potential involvement in diseases.
Used in Microbiology:
In microbiology, 4-Methylumbelliferyl-beta-D-glucuronide is used as a test for coliform bacteria in water. The presence of these bacteria can indicate contamination, and the substrate helps in their identification through the fluorescence emitted upon cleavage.
Used in Microbial Culture Media:
4-Methylumbelliferyl-beta-D-glucuronide is widely used as a component of selective microbial culture media. It serves as a substrate for β-glucuronidase-producing microorganisms, aiding in their isolation and identification.
Used in Chemical Synthesis:
4-Methylumbelliferyl β-D-glucuronide dihydrate is a useful building block and a fluorophore in chemical synthesis. Its fluorescent properties make it a valuable tool in the development of new compounds and materials with potential applications in various fields, such as pharmaceuticals, diagnostics, and bioimaging.

InChI:InChI=1/C16H16O9/c1-6-4-10(17)24-9-5-7(2-3-8(6)9)23-16-13(20)11(18)12(19)14(25-16)15(21)22/h2-5,11-14,16,18-20H,1H3,(H,21,22)/p-1/t11-,12+,13+,14-,16+/m0/s1

Synthetic route

methyl (4′-methylumbelliferyl 2,3,4-tri-O-acetyl-β-D-glucopyranosid)urinate (101014-65-7) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With water; potassium carbonate In methanol; dichloromethane at 0 - 5℃; for 12h;	89%
With barium hydroxide monohydrate In methanol; water for 4h; Inert atmosphere; Cooling with ice;	47%
With barium hydroxide monohydrate In methanol; water at 0℃; for 4h; Inert atmosphere;	47%
Multi-step reaction with 2 steps 1: NaOMe; MeOH / 1 h / 20 °C 2: 0.068 mmol / LiOH / tetrahydrofuran; H2O / 1 h / 20 °C View Scheme

4-methylumbelliferyl-2,3,4-tri-O-acetyl-β-D-glucopyranosiduronic acid (937018-37-6) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With barium methoxide In methanol at 0 - 5℃; for 24h;	85%

7-hydroxy-4-methyl-chromen-2-one (90-33-5, 79566-13-5) + UDP-glucuronic acid (2616-64-0) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With UGT88D4 In aq. buffer at 30℃; for 12h; Reagent/catalyst; Solvent; Enzymatic reaction;	75%
With rat liver microsome solution; magnesium chloride In various solvent(s) at 37℃; for 0.166667h; pH=7.4;

4-methylumbelliferyl β-D-glucopyranoside (18997-57-4) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With oxygen; sodium hydrogencarbonate; platinum at 90℃;
Multi-step reaction with 5 steps 1: pyridine / 5 h / 90 - 100 °C 2: 7.60 g / pyridine / 24 h / 20 °C 3: 92 percent / iodine; methanol / benzene / 24 h / 65 °C 4: 68 percent / aqueous sodium hypochlorite; TBAB; NaBr / 2,2,6,6-tetramethyl-1-piperidinyloxy; tetrabutylammonium bromide / aq. NaHCO3; CH2Cl2 / 3 h / 0 °C 5: 85 percent / barium methoxide / methanol / 24 h / 0 - 5 °C View Scheme

methyl 4-methylcoumarin-7-yl-β-D-glucopyranuronate (116523-82-1) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With lithium hydroxide In tetrahydrofuran; water at 20℃; for 1h;	0.068 mmol

4-methylumbelliferyl-2,3,4-tri-O-acetyl-β-glucopyranoside (937018-36-5) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
Multi-step reaction with 2 steps 1: 68 percent / aqueous sodium hypochlorite; TBAB; NaBr / 2,2,6,6-tetramethyl-1-piperidinyloxy; tetrabutylammonium bromide / aq. NaHCO3; CH2Cl2 / 3 h / 0 °C 2: 85 percent / barium methoxide / methanol / 24 h / 0 - 5 °C View Scheme

4-methyl-7-(3,4,5-trihydroxy-6-trityloxymethyl-tetrahydro-pyran-2-yloxy)-chromen-2-one → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
Multi-step reaction with 4 steps 1: 7.60 g / pyridine / 24 h / 20 °C 2: 92 percent / iodine; methanol / benzene / 24 h / 65 °C 3: 68 percent / aqueous sodium hypochlorite; TBAB; NaBr / 2,2,6,6-tetramethyl-1-piperidinyloxy; tetrabutylammonium bromide / aq. NaHCO3; CH2Cl2 / 3 h / 0 °C 4: 85 percent / barium methoxide / methanol / 24 h / 0 - 5 °C View Scheme

4-methylumbelliferyl-2,3,4-tri-O-acetyl-6-O-trityl-β-D-glucopyranoside (937018-35-4) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
Multi-step reaction with 3 steps 1: 92 percent / iodine; methanol / benzene / 24 h / 65 °C 2: 68 percent / aqueous sodium hypochlorite; TBAB; NaBr / 2,2,6,6-tetramethyl-1-piperidinyloxy; tetrabutylammonium bromide / aq. NaHCO3; CH2Cl2 / 3 h / 0 °C 3: 85 percent / barium methoxide / methanol / 24 h / 0 - 5 °C View Scheme

sodium 4-methylumbelliferonate (5980-33-6) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
Multi-step reaction with 3 steps 1: 45 percent / benzyltriethylammonium chloride; aq. NaOH / CHCl3 / 5 h / Heating 2: NaOMe; MeOH / 1 h / 20 °C 3: 0.068 mmol / LiOH / tetrahydrofuran; H2O / 1 h / 20 °C View Scheme

1-bromo-2,3,4-tri-O-acetyl-α-D-glucuronic acid methyl ester (21085-72-3) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
Multi-step reaction with 3 steps 1: 45 percent / benzyltriethylammonium chloride; aq. NaOH / CHCl3 / 5 h / Heating 2: NaOMe; MeOH / 1 h / 20 °C 3: 0.068 mmol / LiOH / tetrahydrofuran; H2O / 1 h / 20 °C View Scheme

7-hydroxy-4-methyl-chromen-2-one (90-33-5, 79566-13-5) + uridine-5'-diphospho-α-D-glucuronic acid trisodium salt (63700-19-6) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With magnesium chloride In phosphate buffer at 37℃; for 1.25h; pH=7.4; Enzyme kinetics; Further Variations:; Solvents;
With recombinant human UDP-glucuronosyltransferase isoform 1A7; magnesium chloride In dimethyl sulfoxide pH=7.4; Kinetics; Reagent/catalyst; Enzymatic reaction;

7-hydroxy-4-methyl-chromen-2-one (90-33-5, 79566-13-5) + uridine diphosphate glucuronic acidammonium salt → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With magnesium(II) chloride hexahydrate; recombinant human uridine 5′-diphosphate-glucuronosyltransferase 1A9; bovine serum albumin In aq. phosphate buffer at 37℃; for 1h; pH=7.4; Kinetics; Time; Concentration; Reagent/catalyst; Darkness; Enzymatic reaction;

7-hydroxy-4-methyl-chromen-2-one (90-33-5, 79566-13-5) + α-D-glucuronyl fluoride (777038-38-7) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
With Escherichia coli β-glucuronidase E504G mutant In aq. phosphate buffer; tert-butyl alcohol at 37℃; for 48h; pH=7.5; Enzymatic reaction;

7-hydroxy-4-methyl-chromen-2-one (90-33-5, 79566-13-5) + (2S,3S,4S,5R,6S)-3,4,5,6-Tetraacetoxy-tetrahydro-pyran-2-carboxylic acid methyl ester (7355-18-2) → 4-methylumbelliferyl-β-D-glucuronide (6160-80-1)

Conditions	Yield
Multi-step reaction with 2 steps 1: boron trifluoride diethyl etherate; dmap / 1,2-dichloro-ethane / 5 h / 60 °C / Inert atmosphere 2: barium hydroxide monohydrate / water; methanol / 4 h / Inert atmosphere; Cooling with ice View Scheme
Multi-step reaction with 2 steps 1: boron trifluoride diethyl etherate; dmap / 1,2-dichloro-ethane / 5 h / 60 °C / Inert atmosphere 2: barium hydroxide monohydrate / water; methanol / 4 h / Inert atmosphere; Cooling with ice View Scheme
Multi-step reaction with 2 steps 1: triethylamine; boron trifluoride diethyl etherate / dichloromethane / 72 h / 20 °C / Inert atmosphere 2: barium hydroxide monohydrate / water; methanol / 4 h / 0 °C / Inert atmosphere View Scheme

diazomethane (186581-53-3, 908094-01-9) + acetic anhydride (108-24-7) + 4-methylumbelliferyl-β-D-glucuronide (6160-80-1) → methyl (4′-methylumbelliferyl 2,3,4-tri-O-acetyl-β-D-glucopyranosid)urinate (101014-65-7)

Conditions	Yield
With pyridine; diethyl ether Reaktion ueber zwei Stufen;

4-methylumbelliferyl-β-D-glucuronide (6160-80-1) → 7-hydroxy-4-methyl-chromen-2-one (90-33-5, 79566-13-5)

Conditions	Yield
With ethylenediaminetetraacetic acid; sodium acetate; Triton X-100; β-glucuronidase in human liver homogenate at 37℃; for 0.5h; pH=5; Enzyme kinetics; Further Variations:; Catalysts;

UDP disodium + 4-methylumbelliferyl-β-D-glucuronide (6160-80-1) → 7-hydroxy-4-methyl-chromen-2-one (90-33-5, 79566-13-5)

Conditions	Yield
With magnesium(II) chloride hexahydrate; recombinant human uridine 5′-diphosphate-glucuronosyltransferase 1A9; bovine serum albumin In aq. phosphate buffer at 37℃; for 1h; pH=7.4; Kinetics; Time; Reagent/catalyst; Darkness; Enzymatic reaction;

methanol (67-56-1) + 4-methylumbelliferyl-β-D-glucuronide (6160-80-1) → methyl 4-methylcoumarin-7-yl-β-D-glucopyranuronate (116523-82-1)

Conditions	Yield
With sulfuric acid at 65℃; for 1h;

4-methylumbelliferyl-β-D-glucuronide (6160-80-1) → D-glucuronic acid (489-91-8, 528-16-5, 552-12-5, 577-46-8, 643-33-4, 2240-07-5, 6294-16-2, 10133-02-5, 18402-78-3, 18968-14-4, 21179-08-8, 23018-83-9, 33599-45-0, 33599-46-1, 46171-67-9, 46171-69-1, 46172-26-3, 70021-34-0, 71031-08-8, 104195-06-4, 106499-29-0, 124817-72-7) + 6-hydroxy-4-methyl-chromen-2-one (2373-31-1)

Conditions	Yield
With β-glucuronidase In aq. phosphate buffer pH=7.4; Concentration; Enzymatic reaction;

Upstream product

• 18997-57-4 4-methylumbelliferyl β-D-glucopyranoside 
• 90-33-5 7-hydroxy-4-methyl-chromen-2-one 
• 63700-19-6 uridine-5'-diphospho-α-D-glucuronic acid trisodium salt 
• 7355-18-2 (2S,3S,4S,5R,6S)-3,4,5,6-Tetraacetoxy-tetrahydro-pyran-2-carboxylic acid methyl ester 
• 777038-38-7 α-D-glucuronyl fluoride 
• 21085-72-3 1-bromo-2,3,4-tri-O-acetyl-α-D-glucuronic acid methyl ester 
• 101014-65-7 methyl (4′-methylumbelliferyl 2,3,4-tri-O-acetyl-β-D-glucopyranosid)urinate 
• 5980-33-6 sodium 4-methylumbelliferonate 
• 937018-35-4 4-methylumbelliferyl-2,3,4-tri-O-acetyl-6-O-trityl-β-D-glucopyranoside 
• 937018-36-5 4-methylumbelliferyl-2,3,4-tri-O-acetyl-β-glucopyranoside 
• 937018-37-6 4-methylumbelliferyl-2,3,4-tri-O-acetyl-β-D-glucopyranosiduronic acid 
• 116523-82-1 methyl 4-methylcoumarin-7-yl-β-D-glucopyranuronate 
• 2616-64-0 UDP-glucuronic acid 

Downstream Products

• 90-33-5 7-hydroxy-4-methyl-chromen-2-one 
• 489-91-8 D-glucuronic acid 
• 2373-31-1 6-hydroxy-4-methyl-chromen-2-one 
• 116523-82-1 methyl 4-methylcoumarin-7-yl-β-D-glucopyranuronate 

Relevant articles and documents

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A new synthetic route to 4-methylumbelliferyl-β-D-glucopyranosiduronic acid (MUG)

A synthetic route to prepare 4-methylumbelliferyl-β-D- glucopyranosiduronic acid (MUG) from 4-methylumbelliferyl-β-D- glucopyranoside (MUGluc) was developed. The primary hydroxyl group in MUGluc was protected by tritylation followed by acetylation of secondary hydroxyls. The triphenylmethyl group was selectively removed by treatment with iodine-methanol in benzene and the free hydroxyl was transformed into the carboxylic acid by phase-transfer oxidation with sodium hypochlorite and TEMPO as catalyst. Finally, the acetate groups were removed by reaction with barium methoxide in methanol to afford the MUG with an overall yield of 37% from the MUGluc. Georg Thieme Verlag Stuttgart.

Synthesis method for Beta-glucuronidase precipitation type fluorometric substrate

The invention discloses a synthesis method for a Beta-glucuronidase precipitation type fluorometric substrate based on 2-(benzothiazole-2'-yl)-4-bromophenol. The synthesis method comprises the following three steps of reaction: (1) glycosylation reaction; (2) aromatic cyclization reaction; (3) protecting group removal reaction. The yield of each step of reaction in the synthesis method can reach the medium level or more and even up to more than 90 percent, the reaction conversion rate of materials with high prices or preparation costs is relatively high, the total yield of the three steps canreach 37 percent, moreover, the reaction condition is mild, and the synthesis method is easy to implement. Furthermore, the step of glycosylation reaction and the step of protecting group removal reaction in the synthesis method can be applied.

An improved helferich method for the α/β-stereoselective synthesis of 4-methylumbelliferyl glycosides for the detection of microorganisms

An improved Helferich method is presented. It involves the glycosylation of 4-methyl-umbelliferone with glycosyl acetates in the presence of boron trifluoride etherate combined with triethylamine, pyridine, or 4-dimethylaminopyridine under mild conditions, followed by deprotection to give fluorogenic 4-methylumbelliferyl glycoside substrates. Due to the use of base, the glycosylation reaction proceeds more easily, is uncommonly α- or β-stereoselective, and affords the corresponding products in moderate to excellent yields (51%-94%) under appropriate conditions.