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nchsls:c:biochemistry:document:dbil

Direct bilirubin examination procedure

referencesDirect Billirubin reference literature
Direct Billirubin reference literature
Direct Billirubin referance literature
Note: When a new lot is opened, verify kit literature with the uploaded version.
Upload newer version when required and make necessary changes in procedure and process
environmental and safety controlsMSDS of HCL
MSDS of sodium nitrite
MSDS of sulfanilicacid
Preparation of TBIL and DBIL Reagenttbil_dbil_reagent_.ods
The examination is done on Erba XL-640. It is also done on Erbachem when Erba XL-640 is damaged and repair may take longer

Erba XL-640

Erbachem

Note: Following instruction is useful only if operator is familiar with Erbachem

ParameterValue
Sample volume(ul)50
Reagent volume(ul)400/100
Filter546 nm
Aspitation Volume(ul)300
Type of ReactionSample blank
Decimal place1
Calibration, Result calculation, Quality control and sample examination in Erbachem
Prepare for calibration as below every monthly or there is relevent NC. Refer Calibrator and QC
testing materialReplicates
Water blank2
Normal Range Calibrator4
Abnormal range Calibrator4
Normal range QC sera4
Abnormal Range QC sera4
  1. Run all testing material as sample (as described below at Sample Examination)
  2. Evaluate calibration and QC absorbances in spreadsheet. Compare with past results.
  3. If results are acceptable, enter the factor in the erbachem and set decimal place to zero.

sample examination

  1. Check aspiration volume of 300 microliter in test parameter of Direct Billirubin.
  2. Run 1 Randox Human assy control-2(QC 5),1 Randox Human assy control-3(QC 8) in each and every batch.See Assigned values of calibrators and target values of QC here
  3. When instrument asks “Aspirate water”, aspirate water.
  4. When instument ask for reagent blank, press 'No' (To prevent automatic correction of factor)
Note:As for sample blanking two eppendroff cups should be taken for each sample and labelled as test(T) and blank(B) and following should be done.

BLANK­

  1. Take 400 microliter reagent R1 in ependroff cup.
  2. Add 50 microliter sample to it.
  3. Incubate at room temperature for 10 minutes.
  4. Add 100 microliter DIAZO A onlyafter 10 minutes.
  5. When instument ask for sample blank, aspirate it.

TEST

  1. Take 400 microliter reagent R1 in ependroff cup.
  2. Add 50 microliter sample to it.
  3. Incubate at room temperature for 10 minutes.
  4. Add 100 microliter R2 (Diazo A+Diazo B) after 10 minutes.
  5. Take reading at 546nm
  6. Evaluate QC results. If QC results show major deviation from past, rerun batch.
  7. If result comes more than 25mg/dl then dilute the sample about 1:1 with DI water and rerun the the sample.
  8. instructions for determining quantitative results when a result is not within the measurement interval: Dilution of sample with DI water is done in same way as done by Erba XL-640 automatically.
nchsls/c/biochemistry/document/dbil.txt · Last modified: 2022/08/13 07:34 (external edit)