A highly sensitive enzyme-linked immunosorbent assay for the determination of tacrolimus in atopic dermatitis patients.
A highly sensitive enzyme-linked immunosorbent assay method has been developed for the determination of tacrolimus in human blood samples. The assay is a modification of the previously published assay with improved sensitivity. Following extraction of tacrolimus with methanol and sulfosalicylic acid, the samples are incubated for 2 h at room temperature on a Nunc Maxisorb plate that has been treated for nonspecific binding by precoating with polyclonal antibody. The analysis of human blood following the standard addition of tacrolimus (0.02-5.0 ng/ml) demonstrated excellent precision and accuracy over a 6-day period. The interday and intraday co-efficients of variation were 6.0-28.9 and 3.9-15.2%, respectively. The limit of quantitation was 0.05 ng/ml. The method was used to quantitate blood concentration of tacrolimus in patients following the administration of tacrolimus ointment.
Alak
A M
AM
Fujisawa Research Institute of America, Inc., Northwestern University/Evanston Research Park, IL 60201, USA.
Cook
M
M
Bekersky
I
I
D000328:Adult; D002648:Child; D003876:Dermatitis, Atopic / Q000097:blood* / Q000188:drug therapy; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007166:Immunosuppressive Agents / Q000097:blood* / Q000627:therapeutic use; D009824:Ointments; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016559:Tacrolimus / Q000097:blood* / Q000627:therapeutic use
D007166:Immunosuppressive Agents; D009824:Ointments; D016559:Tacrolimus