======Direct bilirubin examination procedure======
  *Learn operation of [[Erba XL-640]] and [[Erbachem]].
  *Select equipment
    *Use Erba XL-640 as peferred equipment.
    *If Erba XL-640 is malfunctioning and can not be repaired within period acceptable to clinicians, use Erbachem and follow [[procedure for responding to Erba XL-640 failure]]
  *Read following documents


^references|{{bilirubin_reference_literature_1.pdf|Direct Billirubin reference literature}}|
| |{{bilirubin_reference_literature_2.pdf|Direct Billirubin reference literature}}|
| |{{bilirubin_reference_literature_3.pdf|Direct Billirubin referance literature}}|
| |**Note:** When a new lot is opened, verify kit literature with the uploaded version.|
| |[[lis_upload_dokuwiki_media|Upload]] newer version when required and make necessary changes in procedure and process|

|**environmental and safety controls**|{{msds of Hcl.pdf|MSDS of HCL}}| 
| |{{msds_sodiumnitrate.pdf|MSDS of sodium nitrite}}|
| |{{MSDS_sulfanilicacid.pdf|MSDS of sulfanilicacid}}| 

|Preparation of TBIL and DBIL Reagent{{:nchsls:c:biochemistry:document:tbil_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======

|[[Centrifugation]]|
|[[Preparing aliquot]]|
|[[nchsls:c:biochemistry:document:how_to_perform_examination_in_erba_xl-640|Procedure for examination in Erba XL-640]]|
======Erbachem======
**Note:** Following instruction is useful only if operator is familiar with [[Erbachem]]

^Parameter^Value^
^Sample volume(ul)|50|
^Reagent volume(ul)|400/100|
^Filter|546 nm|
^Aspitation Volume(ul)|300|
^Type of Reaction|Sample blank|
^Decimal place|1|


^Calibration, Result calculation, Quality control and sample examination in Erbachem^

^[[nchsls:c:biochemistry:document:internal_quality_control|Internal Quality Control]]^
^[[how_to_do_external_quality_control_on_erba_xl-640|EQA]]^

^Prepare for calibration as below every monthly or there is relevent NC. Refer [[Calibrator and QC]]^
^testing material^Replicates^
|Water blank|2|
|Normal Range Calibrator|4|
|Abnormal range Calibrator|4|
|Normal range QC sera|4|
|Abnormal Range QC sera|4|

  -Run all testing material as sample (as described below at **Sample Examination**)
  -Evaluate calibration and QC absorbances in spreadsheet. Compare with past results.
  -If results are acceptable, enter the factor in the erbachem and set decimal place to zero.

====sample examination====
  -Check aspiration volume of 300 microliter in test parameter of Direct Billirubin.
  -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 [[Calibrator and QC Targets|here]]
  -When instrument asks "Aspirate water", aspirate water.
  -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**­
  - Take 400 microliter reagent R1 in ependroff cup. 			
  - Add 50 microliter sample to it.  			
  - Incubate at room temperature for 10 minutes.			
  - Add 100 microliter DIAZO A onlyafter 10 minutes.
  - When instument ask for sample blank, aspirate it.	

**TEST**
  - Take 400 microliter reagent R1  in ependroff cup.			
  - Add 50 microliter sample to it. 			
  - Incubate at room temperature for 10 minutes.			
  - Add 100 microliter R2 (Diazo A+Diazo B) after 10 minutes.
  - Take reading at 546nm
  - Evaluate QC results. If QC results show major deviation from past, rerun batch.
  - If result comes more than 25mg/dl then dilute the sample about 1:1 with DI water and rerun the the sample.
  - [[lis_calibration_entry|Enter Calibration in LIS]]
  - [[list_qc_entry_manual|Enter QC results manually in LIS]]
  - [[list_result_entry_manual|Enter sample results manually in LIS]]
  - **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.