Charcoal stabilization of phenyl phosphates
First Claim
1. A method for stabilizing a colorless solubilized phenyl phosphate comprising contacting the solubilized phenyl phosphate with a stabilizing amount of charcoal and retaining the charcoal in contact with the solubilized phenyl phosphate to provide a stabilized, solubilized phenyl phosphate having a background absorbance of less than about 0.1 at 405 nm and an activity of about 0.2 OD/min.
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Abstract
The invention discloses methods, compositions and kits for stabilizing a solubilized phenyl phosphate, preferably paranitrophenyl phosphate (PNPP), using charcoal. Also disclosed are methods, compositions and kits for recycling solubilized phenyl phosphate, preferably PNPP, that has an absorbance of less than 0.1 when measured at 405 nm due to non-enzymatic hydrolysis.
12 Citations
44 Claims
- 1. A method for stabilizing a colorless solubilized phenyl phosphate comprising contacting the solubilized phenyl phosphate with a stabilizing amount of charcoal and retaining the charcoal in contact with the solubilized phenyl phosphate to provide a stabilized, solubilized phenyl phosphate having a background absorbance of less than about 0.1 at 405 nm and an activity of about 0.2 OD/min.
- 9. A stabilized, solubilized phenyl phosphate composition comprising (a) a solution comprising phenyl phosphate in a buffer, wherein the solution has not previously been colored due to non-enzymatic hydrolysis, and (b) a stabilizing amount of charcoal, wherein the stabilized, solubilized phenyl phosphate has a background absorbance of less than about 0.1 at 405 nm and an activity of about 0.2 OD/min.
- 17. A ready-to-use enzyme substrate composition comprising colorless solubilized phenyl phosphate, a buffer, and charcoal.
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25. A method of preparing an aqueous liquid substrate system used in phosphatase enzyme determination comprising:
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(a) solubilizing a phenyl phosphate in an aqueous buffered solvent to provide a colorless phenyl phosphate solution; (b) adding a magnesium compound to the colorless phenyl phosphate solution; (c) contacting the colorless phenyl phosphate solution with a stabilizing amount of charcoal; (d) retaining the magnesium compound and the charcoal in the colorless phenyl phosphate solution; and (e) sealing the mixture. - View Dependent Claims (26)
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- 27. A vessel containing a colorless solubilized phenyl phosphate in a basic buffer, wherein the vessel comprises charcoal on the surface of the vessel exposed to the solubilized phenyl phosphate.
- 29. A method for stabilizing a solubilized phenyl phosphate which has been colored due to non-enzymatic hydrolysis comprising contacting the solubilized phenyl phosphate with a stabilizing amount of charcoal and retaining the charcoal in contact with the solubilized phenyl phosphate to provide a stabilized, solubilized phenyl phosphate having a background absorbance of less than about 0.1 at 405 nm and an activity of about 0.2 OD/min for 30 days or more in light at room temperature.
- 37. A stabilized, solubilized phenyl phosphate composition comprising (a) a solution comprising phenyl phosphate in a buffer, wherein the solution has previously been colored due to non-enzymatic hydrolysis, and (b) a stabilizing amount of charcoal, wherein the stabilized, solubilized phenyl phosphate has a background absorbance of less than about 0.1 at 405 nm and an activity of about 0.2 OD/min for 30 days or more in light at room temperature.
Specification