System and methods for bilirubin analysis

Abstract

A method for measuring bilirubin levels in a subject. The method can include the steps of providing a sample to be measured from the subject, wherein the sample comprises bilirubin bound to albumin; adding a release agent to the sample, the release agent configured to release the bound bilirubin from the albumin; measuring electrochemical data of the sample using an electrochemical cell; and determining a total serum bilirubin concentration of the sample using the electrochemical data. This method is capable of providing a simpler, faster, and more robust measurement when compared to traditional bilirubin assay methods.

1 . A method for measuring bilirubin levels in a subject, the method comprising:
 providing a fluid sample to be measured from the subject to a microfluidic device, wherein the fluid sample comprises bilirubin bound to albumin;   contacting the fluid sample and a release agent, the release agent configured to release the bound bilirubin from the albumin;   measuring electrochemical data of the fluid sample using an electrochemical cell; and   determining a total serum bilirubin concentration of the fluid sample using the electrochemical data.   
     
     
         2 . The method as recited in  claim 1 , wherein the release agent is dry formulated, lyophilized, or immobilized within a film. 
     
     
         3 . The method as recited in  claim 1 , wherein the fluid sample is an untreated blood sample of the subject. 
     
     
         4 . The method as recited in  claim 1 , wherein the release agent is cocamidopropyl sulfobetaine, 3-[3-Cholamidopropyl)-dimethylamminonio]-1-propanesulfonate, 3-[3-Cholamidopropyl)-dimethylamminonio]-2-hydroxy-1-propanesulfonate, cocamidopropyl betaine, caffeine, sodium benzoate, sodium acetate, ethylene glycol, hexadecanoic acid, (9Z)-octadec-9-enoic acid, haematin, a sulfonamide, salicylate, sodium dodecyl sulfate, or 4,4′-(1,1-dioxido-3H-2,1-benzoxathiole-3,3-diyl)bis(2,6-dibromophenol). 
     
     
         5 . The method of  claim 1 , wherein the volume of the fluid sample is less than 100 microliters. 
     
     
         6 . A method for measuring bilirubin levels in a subject, the method comprising:
 contacting a blood sample from the subject and a release agent, wherein the blood sample comprises bilirubin bound to albumin and the release agent is configured to release the bound bilirubin from the albumin;   measuring electrochemical data of the blood sample using an electrochemical cell; and   determining concentrations of conjugated bilirubin, unconjugated bilirubin, and total serum bilirubin using the electrochemical data.   
     
     
         7 . The method as recited in  claim 6 , wherein the release agent is dry formulated, lyophilized, or immobilized within a film. 
     
     
         8 . The method as recited in  claim 6 , wherein the release agent is cocamidopropyl sulfobetaine, 3-[3-Cholamidopropyl)-dimethylamminonio]-1-propanesulfonate, 3-[3-Cholamidopropyl)-dimethylamminonio]-2-hydroxy-1-propanesulfonate, cocamidopropyl betaine, caffeine, sodium benzoate, sodium acetate, ethylene glycol, hexadecanoic acid, (9Z)-octadec-9-enoic acid, haematin, a sulfonamide, salicylate, sodium dodecyl sulfate, or 4,4′-(1,1-dioxido-3H-2,1-benzoxathiole-3,3-diyl)bis(2,6-dibromophenol). 
     
     
         9 . The method as recited in  claim 6 , further comprising filtering the blood sample, wherein the filtering occurs prior to measuring the electrochemical data. 
     
     
         10 . The method as recited in  claim 6 , wherein the volume of the blood sample is less than 100 microliters. 
     
     
         11 . A method for measuring bilirubin levels in a subject, the method comprising:
 providing a blood sample to be measured from the subject, wherein the blood sample comprises bilirubin bound to albumin;   contacting the blood sample and a release agent, the release agent configured to release the bound bilirubin from the albumin;   measuring electrochemical data of the blood sample using an electrochemical cell; and   determining concentrations of conjugated bilirubin, unconjugated bilirubin, total serum bilirubin, and albumin using the electrochemical data.   
     
     
         12 . The method as recited in  claim 11 , wherein the release agent is dry formulated, lyophilized, or immobilized within a film. 
     
     
         13 . The method as recited in  claim 11 , wherein the release agent is cocamidopropyl sulfobetaine, 3-[3-Cholamidopropyl)-dimethylamminonio]-1-propanesulfonate, 3-[3-Cholamidopropyl)-dimethylamminonio]-2-hydroxy-1-propanesulfonate, cocamidopropyl betaine, caffeine, sodium benzoate, sodium acetate, ethylene glycol, hexadecanoic acid, (9Z)-octadec-9-enoic acid, haematin, a sulfonamide, salicylate, sodium dodecyl sulfate, or 4,4′-(1,1-dioxido-3H-2,1-benzoxathiole-3,3-diyl)bis(2,6-dibromophenol). 
     
     
         14 . The method as recited in  claim 13 , wherein the release agent is 4,4′-(1,1-dioxido-3H-2,1-benzoxathiole-3,3-diyl)bis(2,6-dibromophenol). 
     
     
         15 . The method as recited in  claim 11 , wherein the electrochemical data is voltammetric data. 
     
     
         16 . The method as recited in  claim 11 , further comprising adding additional agents to the blood sample, wherein the addition of the agents occurs prior to measuring the electrochemical data. 
     
     
         17 . The method as recited in  claim 16 , wherein the additional agents are dry formulated, lyophilized, or immobilized within a film. 
     
     
         18 . The method as recited in  claim 16 , wherein the additional agents include at least one of a surfactant, buffer, salt, or second release agent. 
     
     
         19 . The method as recited in  claim 11 , wherein the temperature of the blood sample is actively controlled between 20 and 40 degrees Celsius while the step of measuring the electrochemical data is occurring. 
     
     
         20 . The method as recited in  claim 11 , wherein the volume of the blood sample is less than 100 microliters.