======Total 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|Total Billirubin referance literature}}|
| |{{bilirubin_reference_literature_2.pdf|Total Billirubin referance literature}}|
| |{{bilirubin_reference_literature_3.pdf|Total 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_caffine.pdf|MSDS of caffine}}| 
| |{{msds_EDTA.pdf|MSDS of EDTA}}|
| |{{msds_sodiumacetate.pdf|MSDS of sodium acetate}}| 
| |{{msds_sodiumnitrate.pdf|MSDS of sodium nitrite}}|
| |{{MSDS_sodium banzoate.pdf|MSDS of sodium banzoate}}|
| |{{MSDS_sulfanilicacid.pdf|MSDS of sulfanilicacid}}| 

|Preparation of{{:nchsls:c:biochemistry:document:tbil_dbil_reagent_.ods|}}|Tbill and Dbill reagent}}|
|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]]| 
|[[ Intereference by GOD POD Glucose Reagent]]| 
======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|
^Reading time |5 sec|


^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|
**Note:** ====Always Reconstitute and use fresh cal 2 and cal 3 for calibration of billirubin reagent.====
  -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 Total 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. 
  - |**Note:**For setting the test sequence of TBIl and DBIL, it should always be kept in mind that these two parameters should always be kept before phosphorus and calcium in the "TEST SEQUENCE" format as it would minimize the inference of phosphate buffer from glucose reagent, which earlier had been interfering with phosphorus.|