Reagents and methods for detecting analytes
A reagent for detecting an analyte comprises a flavoprotein enzyme, a mediator such as a phenothiazine mediator, at least one surfactant, a polymer and a buffer. The reagent may be used with an electrochemical test sensor that includes a plurality of electrodes.
1. A reagent for determining an analyte concentration, the reagent comprising:
a flavoprotein enzyme;
a 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator;
at least one surfactant;
a hydroxyethyl cellulose polymer; and
a buffer,
wherein at least one of the mediator and the buffer includes an inorganic salt, and a ratio of the total inorganic salt to mediator is less than about 3:1.
2. The reagent of claim 1 , wherein the flavoprotein enzyme is FAD-glucose dehydrogenase.
3. The reagent of claim 1 , wherein the surfactant includes a saccharide-based surfactant or a phosphorylcholine-based surfactant.
4. The reagent of claim 1 , wherein the buffer includes a phosphate buffer.
5. The reagent of claim 1 , wherein the ratio of the total inorganic salt to mediator is in a ratio of less than about 2:1.
6. A reagent for determining an analyte concentration in a fluid sample, the reagent comprising:
FAD-glucose dehydrogenase having an activity of from about 0.1 Units/μL to about 10 Units/μL;
a 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator having a concentration of from about 5 mM to about 120 mM;
a surfactant having a concentration of from about 0.05 wt. % to about 0.5 wt. % of the reagent;
a hydroxyethyl cellulose polymer having a concentration of from about 0.1 wt. % to about 4 wt. % of the reagent; and
a buffer,
wherein at least one of the mediator and the buffer includes an inorganic salt, and a ratio of the total inorganic salt to mediator is less than about 2:1.
7. The reagent of claim 6 , wherein the buffer includes a phosphate buffer.
8. The reagent of claim 6 , wherein the surfactant is a heptanoyl-N-methylglucamide surfactant.
9. The reagent of claim 6 , wherein the FAD-glucose dehydrogenase has an activity of from about 0.5 Units/μL to about 2.5 Units/μL and the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator has a concentration of from about 30 mM to about 60 mM, the surfactant being a heptanoyl-N-methylglucamide surfactant having a concentration of from about 0.1 wt. % to about 0.4 wt. % of the reagent, and wherein the hydroxyethyl cellulose polymer has a concentration of from about 0.2 wt. % to about 0.5 wt. % of the reagent.
10. A method of determining an analyte concentration in a fluid sample, the method comprising the acts of:
providing an electrode surface;
providing a reagent including a flavoprotein enzyme, a 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator, at least one surfactant, a buffer, and a hydroxyethyl cellulose polymer, the reagent being in contact with the electrode surface;
contacting the fluid sample with the reagent; and
determining the concentration of the analyte,
wherein at least one of the mediator and the buffer includes an inorganic salt, the ratio of the total inorganic salt to mediator being less than about 3:1.
11. The method of claim 10 , wherein the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator includes about 5 wt. % sulfate.
12. The method of claim 10 , wherein the flavoprotein is FAD-glucose dehydrogenase.
13. The method of claim 10 wherein the buffer is a phosphate buffer.
14. The reagent of claim 1 , wherein the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator includes about 5 wt. % sulfate.
15. The reagent of claim 2 , wherein the reagent includes about 2.00 U/μL of the FAD-glucose dehydrogenase enzyme, about 40 mM of the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator, about 50 mM of the buffer, about 0.20 wt. % of heptanoyl-N-methylglucamide surfactant, and about 0.25 wt. % of the hydroxyethyl cellulose polymer.
16. The reagent of claim 4 , the reagent including about 30 mM of the phosphate buffer.
17. The reagent of claim 2 , wherein the FAD-glucose dehydrogenase has an activity of from about 0.5 Units/μL to about 2.5 Units/μL, the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator has a concentration of from about 30 mM to about 60 mM, the surfactant being a heptanoyl-N-methylglucamide surfactant having a concentration of from about 0.1 wt. % to about 0.4 wt. % of the reagent, and the hydroxyethyl cellulose polymer has a concentration of from about 0.2 wt. % to about 0.5 wt. % of the reagent.
18. The reagent of claim 2 , wherein the FAD-glucose dehydrogenase has an activity of about 1.75 Units/μL, the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator has a concentration of about 40 mM, the surfactant is heptanoyl-N-methylglucamide surfactant having a concentration of about 0.2 wt. %, the hydroxyethyl cellulose has a concentration of about 0.25 wt. %, and the buffer is a phosphate buffer having concentration of about 100 mM and a pH of about 6.5, the coefficient of variation of the reagent being between about 1.2 and about 3.4.
19. The reagent of claim 2 , wherein the FAD-glucose dehydrogenase has an activity of about 1.75 Units/μL, the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator has a concentration of about 40 mM, the surfactant having a concentration of about 0.2 wt. %, the hydroxyethyl cellulose has a concentration of about 0.25 wt. %, and the buffer being a phosphate buffer having a concentration of about 100 mM.
20. The reagent of claim 2 , wherein the buffer is a phosphate buffer, the reagent including about 50 mM of the phosphate buffer at a pH of about 6.5, about 0.25% of the hydroxyethyl cellulose-300k polymer, about 0.2% of heptanoyl-N-methylglucamide surfactant, and about 40 mM of the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator having 5 wt. % sulfate, wherein the FAD-glucose dehydrogenase has an activity of about 2 Units/μL, and wherein the coefficient of variation of the reagent is between about 1.4 and about 2.7.
21. The reagent of claim 6 , wherein the 3-(2′,5′-disulfophenylimino)-3H-phenothiazine mediator includes about 5 wt. % sulfate.