- Take 5 ml Qc water(DI water of 0.00 microsiemens/cm) in volumetric flask.
  - Gently transfer water in cal 2 and cal 3 bottle.Pour last drop of water by simply rotate flask over edge of bottle.Do not beat the flask over bottle.
  - Mix it by gentle inversion and eversion.
  - Put it at room temperature for 30 minute and watch for all powder particle has been dissolved.
  - Fill 10 ependroff cup of 500 microliter.and seal with it parafilm.
  - Use fresh Cal 2, Cal 3, QC 5 and QC 8 for bilirubin reagent preparation because bilirubin is degraded gradually when remain for prolong storage. **Don't use stored and frozen calibrators and QC**.