4264-83-9

Basic information

• Product Name: Disodium 4-nitrophenylphosphate
• Synonyms: DisodiuM 4-Nitrophenylphosphate Hexahydrate [for BiocheMical Research];PNPP-NA2;4-Nitrophenyl phosphate, disodiuM salt, hexahydrate, 98+% 10GR;PNPP-NA2/4-NPP;DisodiuM 4-nitrophenyIphosphate;sodiuM 4-nitrophenyl phosphate;para-nitrophenylphosphate (pnpp)(disodiuM salt );PNPPDisodiuM 4-nitrophenylphosphate
• CAS NO: 4264-83-9
• Molecular Formula: C₆H₄NO₆P*2Na
• Molecular Weight: 263.05
• EINECS: 224-246-5
• Product Categories: Substrates;Enzyme substrates;Biological Buffers;Buffer Convenience Packaging;Buffer SolutionsBiological Buffers;Buffers A to Z
• Mol File: 4264-83-9.mol
• Article Data: 4

Chemical Properties

• Melting Point: >300 °C(lit.)
• Boiling Point: 457.849 °C at 760 mmHg
• Flash Point: 230.699 °C
• Appearance: white to pale yellow/tablets
• Density: 1.712g/cm3
• Vapor Pressure: 15Pa at 20-50℃
• Storage Temp.: 2-8°C
• Solubility: H2O: 1 tablet/3 mL 25 mM, clear, light yellow
• Water Solubility: Miscible with water. Immiscible with ethanol and acetone.
• Stability: Stable, but may be light sensitive. Incompatible with strong acids, strong bases, strong oxidizing agents.
• BRN: 5324661
• CAS DataBase Reference: Disodium 4-nitrophenylphosphate(CAS DataBase Reference)
• NIST Chemistry Reference: Disodium 4-nitrophenylphosphate(4264-83-9)
• EPA Substance Registry System: Disodium 4-nitrophenylphosphate(4264-83-9)

Safety Data

• Hazard Codes: Xi,Xn
• Statements: 36/38-36/37/38-48/22-41-38-21/22
• Safety Statements: 26-36-24/25-23-36/37/39
• WGK Germany: 3
• F: 10-21
• TSCA: Yes
• Hazardous Substances Data: 4264-83-9(Hazardous Substances Data)

Usage

Chemical Properties

White crystalline

Uses

PNPP, Disodium Salt is a colorimetric alkaline phosphatase soluble substrate, p-Nitrophenyl Phosphate (pNPP) is the substrate of choice for use with alkaline phosphatase in Enzyme Linked Immunosorbant Assay (ELISA) procedures.

General Description

Excellent substrate for alkaline phosphatase-based ELISA assays. Produces a yellow soluble end product that can be read at 405 nm to 410 nm. Both kinetic and endpoint determinations can be performed. ε max: ≥9600 cm-1 M-1 at 311 nm in 0.4 M Na2CO3.

Biochem/physiol Actions

Primary TargetSubstrate for phosphatase

Purification Methods

Dissolve it in hot aqueous MeOH, filter and precipitate it by adding Me2CO. Wash the solid with Me2CO and repeat the purification. Aqueous MeOH and Et2O can also be used as solvents. The white fibrous crystals contain less than 1% of free 4-nitrophenol [assay: Axelrod J Biol Chem 167 57 1947]. [Beilstein 6 IV 1327.]

InChI:InChI=1/C6H6NO6P.2Na/c8-7(9)5-1-3-6(4-2-5)13-14(10,11)12;;/h1-4H,(H2,10,11,12);;/q;2*+1/p-2

Synthetic route

sodium bis(4-nitrophenyl) phosphate (4043-96-3) → disodium (4-nitrophenyl)phosphate (4264-83-9)

Conditions	Yield
{[Cu{[(1-vinylIm-2-yl)CH2]3N}Cl]PF6}-{[CH2=C(CH3)CO2CH2-]n} In water; dimethyl sulfoxide at 50℃; for 96h; pH=9.5; Kinetics; Further Variations:; Catalysts; pH-values; Temperatures; catalyst ratios;	53%

C6H4NO6P(2-)*C30H57N6(3+)*2Cl(1-)*Na(1+) → C30H57N6(3+)*3Cl(1-) + disodium (4-nitrophenyl)phosphate (4264-83-9)

Conditions	Yield
With sodium chloride In d(4)-methanol at 65℃; Equilibrium constant;

sodium bis(4-nitrophenyl) phosphate (4043-96-3) → 4-nitro-phenol (100-02-7) + disodium (4-nitrophenyl)phosphate (4264-83-9)

Conditions	Yield
With [D]-sodium hydroxide; (Et2NH2)8[{α-PW11O39Zr(μ-OH)(H2O)}2]*7H2O; sodium diphenyl phosphate; hydrogen chloride In water-d2 at 60℃; pH=6.4; Kinetics; Reagent/catalyst; Temperature; Concentration;

4-nitro-phenol (100-02-7) → disodium (4-nitrophenyl)phosphate (4264-83-9)

Conditions	Yield
Multi-step reaction with 4 steps 1: triethylamine / dichloromethane / 4 h / 20 °C 2: acetonitrile / 16 h / 0 - 20 °C 3: water / methanol / 1 h / 0 - 20 °C 4: sodium hydroxide / water / 0 °C / Cooling with ice View Scheme
Multi-step reaction with 4 steps 1: triethylamine / dichloromethane / 4 h / 20 °C 2: acetonitrile / 24 h / 0 - 20 °C 3: water / methanol / 1 h / 0 - 20 °C 4: sodium hydroxide / water / 0 °C / Cooling with ice View Scheme

paraoxon (311-45-5) → disodium (4-nitrophenyl)phosphate (4264-83-9)

Conditions	Yield
Multi-step reaction with 3 steps 1: acetonitrile / 16 h / 0 - 20 °C 2: water / methanol / 1 h / 0 - 20 °C 3: sodium hydroxide / water / 0 °C / Cooling with ice View Scheme
Multi-step reaction with 3 steps 1: acetonitrile / 24 h / 0 - 20 °C 2: water / methanol / 1 h / 0 - 20 °C 3: sodium hydroxide / water / 0 °C / Cooling with ice View Scheme

C12H22NO6PSi2 → disodium (4-nitrophenyl)phosphate (4264-83-9)

Conditions	Yield
Multi-step reaction with 2 steps 1: water / methanol / 1 h / 0 - 20 °C 2: sodium hydroxide / water / 0 °C / Cooling with ice View Scheme

mono-p-nitrophenyl phosphate (330-13-2) → disodium (4-nitrophenyl)phosphate (4264-83-9)

Conditions	Yield
With sodium hydroxide In water at 0℃; Cooling with ice;	297 g

[[1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato]dizinc(II)(CH3COO)][ClO4]2*H2O + disodium (4-nitrophenyl)phosphate (4264-83-9) → [[1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato]dizinc(II)(p-nitrophenyl phosphate)][ClO4]2*H2O

Conditions	Yield
In water 1:1 mixt. of Zn complex and p-nitrophenyl phosphate disodium salt;	99%

bis(N,N-dimethylformamide)dichloridodioxidomolybdenum(VI) (21356-99-0, 18078-67-6) + disodium (4-nitrophenyl)phosphate (4264-83-9) → 3H(1+)*Mo12O40P(3-)*3C2H7N

Conditions	Yield
With water at 55℃; for 0.5h;	95%

disodium (4-nitrophenyl)phosphate (4264-83-9) → 4-nitro-phenol (100-02-7)

Conditions	Yield
With cerium(IV) oxide In ethanol at 25℃; for 8h; Kinetics;	91%
With tetraethylammonium hydroxide In water; acetonitrile at 20℃;
at 37℃; enzimatically by alkaline and acid phosphatase;

zinc perchlorate + 1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-ol*4HClO4*2.5H2O + disodium (4-nitrophenyl)phosphate (4264-83-9) → [[1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-olato]dizinc(II)(p-nitrophenyl phosphate)][ClO4]2*H2O

Conditions	Yield
With NaOH In water adding aq. soln. of amine to Zn salt, adding aq. NaOH, stirring for 1 h at 45°C, filtration, adding. aq. soln. of p-nitrophenyl phosphatedisodium salt; cooling to room temp.; elem. anal.;	80%

tetraaqua(1,3-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene)dicopper(II) perchlorate trihydrate*NaClO4 + disodium (4-nitrophenyl)phosphate (4264-83-9) → (4-nitrophenyl phosphate)(1,3-bis(1,4,7-triazacyclonon-1-ylmethyl)benzene)(μ-OH)dicopper(II) perchlorate monohydrate

Conditions	Yield
With HEPES buffer; NaClO4 In water soln. of Cu complex, Na2O2P(O)OC6H4NO2, HEPES buffer (pH 7.4) and NaClO4kept at 38°C overnight; crystals collected; washed with water; elem. anal.;	70%

Zn2(C16H32(NH)6N2)(C3H5O)(3+)*3ClO4(1-)=[Zn2(C16H32(NH)6N2)(C3H5O)](ClO4)3 + disodium (4-nitrophenyl)phosphate (4264-83-9) → Zn2(C16H32(NH)6N2)(C3H5O)(HPO3)(3+)*3ClO4(1-)*2H2O=[Zn2(C16H32(NH)6N2)(C3H5O)(HPO3)](ClO4)3*2H2O

Conditions	Yield
With HClO4 In water stirring (60°C), 2 d), adjustment of pH to 3 (HClO4), evapn. of solvent; recrystn. (water); elem. anal.;	69%

zinc(II) perchlorate + 4,16-(2-hydroxypropano)-1,4,7,13,16,19-hexaazacyclotetracosane tetrahydrochloride hemihydrate + water (7732-18-5) + disodium (4-nitrophenyl)phosphate (4264-83-9) → Zn2(OCH(CH2N(CH2CH2NH)2)2((CH2)5)2)(O2P(O)OC6H4NO2)(1+)*ClO4(1-)*4H2O=(Zn2(C21H45N6O)(O2P(O)OC6H4NO2))ClO4*4H2O

Conditions	Yield
With NaOH In water aq. soln. of Zn salt added to aq. soln. of HCl salt of ligand; 0.10 M NaOH (5 equiv.) added dropwise at 45°C for 1 h; soln. filtered withcellulose nitrate filter; Na phosphate added; cooled to room temp.; elem. anal.;	69%

[Co(tris(2-pyridylmethyl)amine)(CO3)]Cl*1.5(water) + disodium (4-nitrophenyl)phosphate (4264-83-9) → [Co(tris(2-pyridylmethyl)amine)(phosphate)] (862125-30-2) + [[Co(tris(2-pyridylmethyl)amine)](μ-diphosphate)](ClO4)2

Conditions	Yield
With charcoal; NaOH In water to a soln. of Co-complex added a soln. of disodium 4-nitrophenylphosphate; mixed soln. adjusted to pH 3 with NaOH soln.; heating to 60°C and catalytic amount of active charcoal added; stirred at 60°C for 7 h; soln. neutralized and products purified by column chromy. with cation-exchange resin;	A 8% B 51%
With charcoal; NaOH In water to a soln. of Co-complex added a soln. of disodium 4-nitrophenylphosphate; mixed soln. adjusted to pH 3 with NaOH soln.; heating to 60°C and catalytic amount of active charcoal added; stirred at 60°C for 5 h; soln. neutralized and products purified by column chromy. with cation-exchange resin;	A n/a B 46%
With charcoal; NaOH In water to a soln. of Co-complex added a soln. of disodium 4-nitrophenylphosphate; mixed soln. adjusted to pH 3 with NaOH soln.; heating to 60°C and catalytic amount of active charcoal added; stirred at 60°C for 3 h; soln. neutralized and products purified by column chromy. with cation-exchange resin;	A n/a B 26%

methanol (67-56-1) + [Cu2(μ-OH)(m-xylenediamine bis(Kemp's triacid imide)(2-))(1,10-phenanthroline)2]NO3*2CH2Cl2 + disodium (4-nitrophenyl)phosphate (4264-83-9) → [Cu2(μ-4-nitrophenylphosphate)(m-xylenediamine bis(Kemp's triacid imide)(2-))(1,10-phenanthroline)2(CH3OH)]*CH3OH

Conditions	Yield
In methanol; dichloromethane soln. disodium 4-nitrophenylphosphate in MeOH-CH2Cl2 was added to soln. Cu complex in MeOH and stirred at room temp. for 12 h; solvent was evapd. in vacuo, residue was extd. with MeOH and concd., Et2O was added; elem. anal.;	28%

disodium (4-nitrophenyl)phosphate (4264-83-9) + methyloxirane (75-56-9, 16033-71-9) → barium 2-hydroxypropyl(4-nitrophenyl)phosphate

Conditions	Yield
Stage #1: disodium (4-nitrophenyl)phosphate; methyloxirane With ammonium hydroxide In water at 35℃; Stage #2: With barium dihydroxide at 20℃; pH=Ca.7;	0.4%

tetraethylammonium hydroxide (77-98-5) + disodium (4-nitrophenyl)phosphate (4264-83-9) → Tentraethylammonium 4-nitrophenoxide (3774-76-3)

Conditions	Yield
With water In acetonitrile at 20℃; Rate constant; Mechanism; Thermodynamic data; ΔH(excit.), ΔS(excit.); other concentration of water , Et4N(1+)*OH(1-) and temperatures;

disodium (4-nitrophenyl)phosphate (4264-83-9) → p-nitrophenolate (14609-74-6)

Conditions	Yield
With water; Zn*(im-bzl-L-His)2dien at 35 - 50℃; Kinetics;
With C91H151N3O70Zn In water at 35℃; pH=6.5 - 9.5; Kinetics; pH-value; Good's buffer;
With [Zn2(2-[bis(2-methoxyethyl)aminomethyl]-4-methylphenol)2Cl2] In water; N,N-dimethyl-formamide Kinetics; Reagent/catalyst; Concentration;

disodium (4-nitrophenyl)phosphate (4264-83-9) → 4-nitro-phenol (100-02-7) + orthophosphate (10percent)

Conditions	Yield
With pentan-1-ol; sodium citrate; erythrocyte acid phosphatase (EAPase) at 37℃; for 0.166667h; Mechanism; pH=5.5, effects of various alcohols on EAPase and Prostatic acid phosphatase (PAPase) enzymes activity, Km, Vmax constants;

[Co(tren)(OH)(H2O)](ClO4)2 + disodium (4-nitrophenyl)phosphate (4264-83-9) → [Co(tris(2-aminoethyl)amine)(O2PO2)] + [Co(tris(2-aminoethyl)amine)(H2O)(OPO3H)](1+) + [Co(tris(2-aminoethyl)amine)(H2O)(OPO3H)](1+) + Co(NH2C2H4)3N(H2O)OPO3Co(NH2C2H4)3NOH2(3+) + Co(NH2C2H4)3N(H2O)OPO3Co(NH2C2H4)3NOH2(3+)

Conditions	Yield
With NaOH In water byproducts: (NH2C2H4)3NCo(O2P(O)O)CoN(C2H4NH2)3(3+), (NH2C2H4)3NCo(OH)(OPO3)]; 40°C, pH from 4.61 to 8.44; not sepd., detected by NMR spectra;

trans-dichlorobis(ethylenediamine)cobalt(III) chloride (13408-72-5, 14040-32-5, 14040-33-6, 20594-10-9, 20594-11-0) + disodium (4-nitrophenyl)phosphate (4264-83-9) → cis-p-nitrophenylphosphatobis(ethylenediamine)cobalt(III) chloride (85436-20-0)

Conditions	Yield
In water mixing into a past using a few drops of water; warming for a few min atca 60°C; diln. with water; heating for 5-10 min at 60-70°C; filtration, washing with water, EtOH, Et2O; drying in vac.; elem. anal.;

[Co(N-2-aminoethyl-N,N-bis(3-aminopropyl)amine)(H2O)(OH)](ClO4)2 + disodium (4-nitrophenyl)phosphate (4264-83-9) → [Co(N-2-aminoethyl-N,N-bis(3-aminopropyl)amine)O2PO2] + Co(NH2C3H6)2NC2H4NH2O2PO2Co(NH2C3H6)2NC2H4NH2(3+) + Co(NH2C3H6)2NC2H4NH2O2PO2Co(NH2C3H6)2NC2H4NH2(3+) + Co(NH2C3H6)2NC2H4NH2O2PO2Co(NH2C3H6)2NC2H4NH2(3+)

Conditions	Yield
With NaOH In water 25°C, pH from 4.88 to 8.01; not sepd., detected by NMR spectra;

[Co(N-2-aminoethyl-N,N-bis(3-aminopropyl)amine)(H2O)(OH)](ClO4)2 + disodium (4-nitrophenyl)phosphate (4264-83-9) → [Co(N-2-aminoethyl-N,N-bis(3-aminopropyl)amine)(OH)O2PO(OC6H4NO2)] + [Co(N-2-aminoethyl-N,N-bis(3-aminopropyl)amine)(OH2)O2PO(OC6H4NO2)](1+)

Conditions	Yield
In water Kinetics; 15°C, pH 4.57; not sepd., detected by NMR spectra;

disodium (4-nitrophenyl)phosphate (4264-83-9) → disodium salt of p-aminophenyl phosphate

Conditions	Yield
With triethanolamine; riboflavin In water at 40℃; for 4h; UV-irradiation;
With hydrogen; palladium 10% on activated carbon In ethanol; water at 20℃; under 1551.49 Torr; for 2h;

sodium methylate (124-41-4) + disodium (4-nitrophenyl)phosphate (4264-83-9) → bis-sodium salt of methyl phosphate (17323-81-8, 51528-83-7)

Conditions	Yield
In methanol; d(4)-methanol at 50℃; Kinetics; Reagent/catalyst; pH-value; Time;

C30H57N6(3+)*3Cl(1-) + disodium (4-nitrophenyl)phosphate (4264-83-9) → C6H4NO6P(2-)*C30H57N6(3+)*2Cl(1-)*Na(1+)

Conditions	Yield
In d(4)-methanol at 65℃; Equilibrium constant;

5,11,17,23-tetra-tert-butyl-25,27-dihydroxy-26,28-bis((R)-α-alanyl-carbonylmethoxy)calix[4]arene + disodium (4-nitrophenyl)phosphate (4264-83-9) → C6H4NO6P(2-)*C54H70N2O10*2Na(1+)

Conditions	Yield
In water; dimethyl sulfoxide at 25℃;

5,11,17,23-tetra-tert-butyl-25,27-dihydroxy-26,28-bis((R)-α-glutyl-carbonylmethoxy)calix[4]arene + disodium (4-nitrophenyl)phosphate (4264-83-9) → C6H4NO6P(2-)*C58H74N2O14*2Na(1+)

Conditions	Yield
In water; dimethyl sulfoxide at 25℃;

disodium (4-nitrophenyl)phosphate (4264-83-9) + 5,11,17,23-tetra-tert-butyl-25,27-dihydroxy-26,28-bis(glycyl-carbonylmethoxy)calix[4]arene → C6H4NO6P(2-)*C52H66N2O10*2Na(1+)

Conditions	Yield
In water; dimethyl sulfoxide at 25℃;

disodium (4-nitrophenyl)phosphate (4264-83-9) + alanine isopropyl ester hydrochloride (39613-92-8, 62062-56-0, 62062-65-1) + triethylamine (121-44-8) → C12H17N2O7P*C6H15N

Conditions	Yield
With dicyclohexyl-carbodiimide In water; tert-butyl alcohol Reflux;

disodium (4-nitrophenyl)phosphate (4264-83-9) → C14H21N2O7PS

Conditions	Yield
Multi-step reaction with 2 steps 1: dicyclohexyl-carbodiimide / tert-butyl alcohol; water / Reflux 2: N,N-dimethyl-formamide / 1 h / 25 °C / Inert atmosphere View Scheme

disodium (4-nitrophenyl)phosphate (4264-83-9) → phosphonic acid mono-(E)-(4-{7-fluoro-6-morpholin-4-yl-2-[2-(5-nitrofuran-2-yl)vinyl]quinazolin-4-ylamino}phenyl) ester disodium salt

Conditions	Yield
Multi-step reaction with 2 steps 1: hydrogen / palladium 10% on activated carbon / ethanol; water / 2 h / 20 °C / 1551.49 Torr 2: N,N-dimethyl-formamide / 3 h / 90 °C View Scheme

Upstream product

• 330-13-2 mono-p-nitrophenyl phosphate 
• 311-45-5 paraoxon 
• 100-02-7 4-nitro-phenol 
• 4043-96-3 sodium bis(4-nitrophenyl) phosphate 

Downstream Products

• 3774-76-3 Tentraethylammonium 4-nitrophenoxide 
• 100-02-7 4-nitro-phenol 
• 17323-81-8 bis-sodium salt of methyl phosphate 
• 824-78-2 4-nitrophenol sodium salt 
• 14609-74-6 p-nitrophenolate 
• 85436-20-0 cis-p-nitrophenylphosphatobis(ethylenediamine)cobalt(III) chloride 

Relevant articles and documents

Enhanced hydrolytic activity of Cu(II) and Zn(II) complexes in highly cross-linked polymers

The chelate ligand tris[(1-vinylimidazol-2-yl)methyl]amine (5) was synthesized in five steps from commercially available starting materials. Upon reaction with ZnCl2 or CuCl2 in the presence of NH 4PF6, the complexes [Zn(5)Cl]PF6 (6) and [Cu(5)Cl]PF6 (7) were obtained. The structure of both complexes was determined by single-crystal X-ray crystallography. Immobilization of 6 and 7 was achieved by co-polymerization with ethylene glycol dimethacrylate. The supported complexes P6-Zn and P7-Cu were found to be efficient catalysts for the hydrolysis of bis(p-nitrophenyl)phosphate (BNPP) at 50 °C. At pH 9.5, the heterogeneous catalyst P7-Cu was 56 times more active than the homogeneous catalyst 7. Partitioning effects, which increase the local concentration of BNPP in the polymer, are shown to contribute to the enhanced activity of the immobilized catalyst. The Royal Society of Chemistry 2006.

Solution speciation of the dinuclear ZrIV-substituted keggin polyoxometalate [{α-PW11O39Zr(μ-OH)(H2O)}2]8- and Its reactivity towards DNA-model phosphodiester hydrolysis

The solution speciation of the ZrIV-substituted Keggin polyoxometalate (Et2NH2)8[{α-PW11O39Zr(μ-OH)(H2O)}2]·7H2O (ZrK 2:2) was fully determined under differ