University of Wisconsin–Madison

Assay Quality Controls

HPLC based QC

  • Blanks
  • Recovery
  • Linerarity
  • LLOQ (lower limits of quantification)
  • LLOD (lower limits of detection)
  • Intra and inter assay coefficient of variation (CV)

Immunoassay based QC

  • Intra and inter assay coefficient of variation (CV)
  • Parallelism
  • Accuracy
  • Sensitivity of the assay

Examples of HPLC based QCs

25-hydroxyvitamin D2 QC Summary for Marmoset Serum samples
LLOD = 0.5 ng/ml, r=0.9999 for calibration curve, blanks = 0
QC Pool ID UTAK Verified Value UTAK Expected Range WNPRC Mean Value MRM1/MRM2 (n=3) WNPRC Mean % Recovery MRM1/MRM2 Std. Dev. MRM1/MRM2 CV MRM1/MRM2
UTAK Low Control 11 9-13 8.4/8.5 76.5/77.0 0.3/0.7 3.1/8.0
UTAK Medium Control 33 28-38 26.3/26.1 79.7/79.0 1.9/1.7 7.1/6.5
UTAK High Control 77 65-89 65.4/64.1 85.0/83.3 2.4/3.2 3.7/4.9

Examples of Immunoassay based QCs

*PdG = pregnanediol-glucuronide
QC pool ID Media Assay type Parallelism Accuracy Sensitivity Intra assay CV % Inter assay CV %
Marmoset serum Insulin RIA t=-1.67, df=26, P > 0.05 98.89+2.38 s.e.m. 0.25 µU/ml 2.87 4.89
Low Rhesus urine *PdG EIA t=-1.77, df=23, P > 0.05 104.01+1.41 s.e.m. 9 pg 4.4 13.9
High Rhesus urine PdG EIA t=-1.77, df=23, P > 0.05 104.01+1.41 s.e.m. 9 pg 3.0 11.6