Sulfanilic acid → 4-[2-[5-[(3-ethenyl-4-methyl-5-oxopyrrol-2-yl)methylidene]-4-methyl-3-propylpyrrol-2-yl]hydrazinyl]benzenesulfonic acid
Reaction equation
CC1C(CCC(O)=O)=C(CC2[NH:22][C:21](/[CH:23]=[C:24]3/[C:25]([CH:31]=[CH2:32])=[C:26]([CH3:30])[C:27]([NH:29]/3)=[O:28])=[C:20](C)[C:19]=2CCC(O)=O)NC=1/C=C1/C(C)=C(C=C)C(N/1)=O.[CH:44]1[C:49]([NH2:50])=[CH:48][CH:47]=[C:46]([S:51]([OH:54])(=[O:53])=[O:52])[CH:45]=1>>[CH3:26][CH2:25][CH2:24][C:23]1[C:21]([NH:22][NH:50][C:49]2[CH:44]=[CH:45][C:46]([S:51]([OH:54])(=[O:52])=[O:53])=[CH:47][CH:48]=2)=[N:22][C:21](=[CH:23][C:24]2[C:25]([CH:31]=[CH2:32])=[C:26]([CH3:30])[C:27](=[O:28])[N:29]=2)[C:20]=1[CH3:19]Participants
3
Procedure
Assays for determination of total bilirubin (serum or plasma) levels may be adapted for use on a droplet actuator. In one example, the total bilirubin assay kit (B576-480) available from Teco Diagnostics may be adapted for use on a droplet actuator. The total bilirubin assay is a colorimetric assay based on the reaction between bilirubin and diazotized sufanilic acid to produce azobilirubin which has an absorbance maximum at 560 nm wavelength in the presence of dimethylsulfoxide (DMSO) solvent. The kit includes the following reagents: Total Bilirubin Reagent (sulfanilic acid 32 mM, hydrochloric acid 165 mM, DMSO 7 M (55% w/v)); and Bilirubin Nitrite Reagent (sodium nitrite 60 mM); Bilirubin Calibrator (20 mg/dL N-1-naphthylethylenediamine dihydrochloride salt). Working reagents and calibration solutions were prepared on-bench as follows: Total bilirubin working reagent was prepared on-bench before each experiment by adding 50 μL of sodium nitrite reagent to 1.0 mL of total bilirubin reagent. Bilirubin calibrator solutions were prepared on-bench at 3, 7, 10, 14, and 20 mg/dL by diluting the 20 mg/dL calibrator stock using deionized water.