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Chemical Structure| 119171-73-2 Chemical Structure| 119171-73-2

Structure of ADHP
CAS No.: 119171-73-2

Chemical Structure| 119171-73-2

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ADHP is a fluorescent peroxidase substrate (λex=530 nm, λem=590 nm).

Synonyms: 10-Acetyl-3,7-dihydroxyphenoxazine; A 6550; Amplex red reagent

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Product Citations

Product Citations

Liu, Huan ; Tonoyan, Lusine ; Reiz, Béla ; Siraki, Arno G ;

Abstract: Amplex Red (AR) is commonly used to detect extracellular hydrogen peroxide (H2O2) and is considered a cellimpermeable compound. However, it would appear capable of entering cells based on its phenoxazine substructure and the report of its mitochondrial membrane permeability. Additionally, myeloperoxidase (MPO) oxidation of AR produces a fluorescent compound, resorufin, which has been reported, though the mechanism is not well-studied. EPR spin trapping using glutathione (GSH) revealed that AR metabolism produced AR radicals and glutathionyl radicals (GS). An intermediate metabolite, 3,7-dihydroxyphenoxazine, was observed by liquid chromatography-mass spectrometry (LC-MS), which supported AR radical disproportionation first and subsequently N-oxidation. Besides, in the presence of GSH, the formation of resorufin decreased significantly evidencing the reactivity of radical intermediates. Three types of AR-GS adduct were found using LC-MS and the resorufin GS-adduct was the dominant one. Regarding intracellular findings in HL-60 cells (that highly express MPO), LC-MS and fluorescence analysis showed AR penetrated the cell membrane and was oxidized by cellular MPO. Interestingly, we demonstrated that the oxidation of AR in HL-60 cells showed a significant time dependence; PF-1355, an MPO inhibitor, inhibited the oxidation of AR by MPO. Cell viability (ATP) revealed that 200 μM AR significantly decreased viability in HL-60 cells in 6 h. We also found that AR-mediated decreased total GSH and increased protein-radical formation. These findings revealed that AR is cell-permeable, and AR radicals induce cellular oxidative distress and lead to the formation of protein radicals, which correlate with the MPO-mediated mechanism of cytotoxicity.

Keywords: Amplex Red ; Myeloperoxidase ; EPR ; Free radicals ; Oxidative stress ; Protein radicals

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Product Details of ADHP

CAS No. :119171-73-2
Formula : C14H11NO4
M.W : 257.24
SMILES Code : CC(N1C2=C(C=C(O)C=C2)OC3=CC(O)=CC=C13)=O
Synonyms :
10-Acetyl-3,7-dihydroxyphenoxazine; A 6550; Amplex red reagent
MDL No. :MFCD00467861

Safety of ADHP

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302
Precautionary Statements:P280-P305+P351+P338

Isoform Comparison

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Bio Calculators
Preparing Stock Solutions 1mg 5mg 10mg

1 mM

5 mM

10 mM

3.89mL

0.78mL

0.39mL

19.44mL

3.89mL

1.94mL

38.87mL

7.77mL

3.89mL

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