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 +|[[clinical_biochemistry_section|Home]]|[[clinical_biochemistry|]]|[[examination_procedures|]]|
  
-' +                      Triglyceride Examination Procedure 
-Purpose of examination:  +**1.Purpose of examination:**  
-●Triglyceride estimation from serum or plasma by Glycerol Phosphate Oxidase method.+Triglyceride estimation from serum or plasma by Glycerol Phosphate Oxidase.
  
-Responsibility and Authority:  +**2.Responsibility and Authority:**  
-● Calibration: Technician  +  * Calibration: Technician  
-● Quality Control: Technician  +  * Quality Control: Technician  
-● Routine operation: Technician  +  * Routine operation: Technician  
-● Overall Monitoring: Quality Manager+  * Overall Monitoring: Quality Manager
  
- Sample Details:  +**3.Sample Details:**  
-● Type of Sample: Serum,  Plasma +  * Type of Sample: Serum,  Plasma 
-● Type of container and additives: Plain without any additives  +  * Type of container and additives: Plain without any additives  
-● Patient Preparation: As per Primary Sample Collection Manual {{Sample collection manual}} +  * Patient Preparation: As per Primary Sample Collection Manual  [[sample_collection_manual|]] 
-● Stability: At Room temperature 18°–28°C (64°–82°F) stability ≤ 24 hours +  * Stability: At Room temperature 18°–28°C (64°–82°F) stability ≤ 24 hours 
-● Handling and transport: As per Primary Sample collection manual  +  * Handling and transport: As per Primary Sample collection manual  
-● Storage: 24 hours at 2-8° C+  * Storage: 24 hours at 2-8° C
  
-Required Equipment:  +**4.Required Equipment:**  
-●Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer+Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer
  
-Required reagents: +**5.Required reagents:** 
-       R1  ATP (2.5 mmol/L),  +  * R1 84ml Active ingredients: ATP (2.5 mmol/L), Mg2+ (2.5 mmol/L), 
-              Mg2+ (2.5 mmol/L), +  * 4-aminoantipyrine (0.4 mmol/L), 4-chlorophenol (2 mmol/L), 
-              4-aminoantipyrine (0.4 mmol/L),  +  * Peroxidase (horseradish) (> 2000 U/L), GK (microbial) (> 600 U/L), 
-              4-chlorophenol (2 mmol/L), +  * GPO (microbial) (> 6000 U/L), Lipoprotein lipase (microbial) (> 3000 U/L).
-              Peroxidase (horseradish) (> 2000 U/L),  +
-              GK (microbial) (> 600 U/L), +
-              GPO (microbial) (> 6000 U/L), +
-              Lipoprotein lipase (microbial) (> 3000 U/L).+
  
- Reagent Handling +**6.Reagent Handling** 
- ● Remove any air bubbles present in the reagents with a new applicator stick, or allow the reagents to settle at the appropriate storage temperature to allow the bubbles to dissipate. To minimize volume depletion, do not use a transfer pipette to remove bubbles +  * Remove any air bubbles present in the reagents with a new applicator stick, or allow the reagents to settle at the appropriate storage temperature to allow the bubbles to dissipate. To minimize volume depletion, do not use a transfer pipette to remove bubbles 
-Reagent Storage and stability +  * Reagent Storage and stability 
-Unopened reagent stable at 2-8°C until expiration date.+  * Unopened reagent stable at 2-8°C until expiration date. 
 +  * On board System temperature reagent is  stable for 30 days 
 +  * Instability or deterioration should be suspected if there are precipitates, visible signs of leakage or 
 +  * contamination, turbidity, or if calibration or controls do not meet the appropriate criteria.
  
-On board System temperature reagent is  stable for 42 days 
-Instability or deterioration should be suspected if there are precipitates, visible signs of leakage or 
-contamination, turbidity, or if calibration or controls do not meet the appropriate criteria. 
  
-Calibration Procedure: 
-● Consolidated Chemistry Calibrator 
  
-● Frequency: +**7.Calibration Procedure:** 
-Reagent lot change +  * Consolidated Chemistry Calibrator 
-QC out of range +  * Frequency: 
-After service or maintenance +  * Reagent lot change 
-Replacement in any parts of Instrument +  * QC out of range 
-Procedure: +  * After service or maintenance 
-Start the equipment.WDI abbotte fully.docx +  * Replacement in any parts of Instrument
-Calibrators are ready to use. +
-Put calibrator 15-20 minutes at room temperature   +
-Prior to use, mix the contents by inverting the vial. Do not shake the vial to prevent foam formation.  +
-Take a 150 µl calibrator solution in separate aliquots. +
-Go to the calibration and give the calibration order. +
-Verify calibration with at least two levels of controls. +
-If control results fall outside acceptable ranges,root cause analysis or recalibration may be necessary.+
  
-Quality control Procedure: +**8.Procedure:** 
-Name: Bio Rad Level 1 & 2 +  - Start the equipment.WDI abbotte fully.docx 
-Frequency: As per Quality Control Procedure +  - Calibrators are ready to use. 
-●Procedure for Reconstitution of IQC  +  - Put calibrator 15-20 minutes at room temperature   
-Reconstitution of QC material with 5 ml Distilled water by using calibrated fixed volume pipette. +  - Prior to use, mix the contents by inverting the vial. Do not shake the vial to prevent foam formation.  
-Leave to stand for 30 min in the dark place. +  - Take a 150 µl calibrator solution in separate aliquots. 
-Swirl gently several times during the reconstitution period to ensure that the contents are completely dissolved. +  - Go to the calibration and give the calibration order. 
-Prior to use, mix the contents by inverting the vial. Do not shake the vial as the information of foam should be avoided. Ensure that no lyophilized material remains un-reconstituted. +  - Verify calibration with at least two levels of controls. 
-Prepare aliquots of 150 µl from the reconstituted QC material. +  - If control results fall outside acceptable ranges,root cause analysis or recalibration may be necessary.
-Store these aliquots at -15° C to -20° C. +
-Prior to use, make sure that aliquots should be at room temperature for at least 15 min.+
  
 +**9.Quality control Procedure:**
 +  * ** Name:** Bio Rad Level 1 & 2
 +  * **Frequency:** As per Quality Control Procedure
  
-●Procedure to run IQC +**Procedure for Reconstitution of IQC**  
-Press Control order  +  - Reconstitution of QC material with 5 ml Distilled water by using calibrated fixed volume pipette. 
-Select Assay /Panel, to be run. +  - Leave to stand for 30 min in the dark place. 
-Select the control/s and its level/s +  - Swirl gently several times during the reconstitution period to ensure that the contents are completely dissolved. 
-Give Carrier Number and Position number +  - Prior to use, mix the contents by inverting the vial. Do not shake the vial as the information of foam should be avoided. Ensure that no lyophilized material remains un-reconstituted. 
-Press F3 / Add order +  - Prepare aliquots of 150 µl from the reconstituted QC material. 
-Put respected carrier in RSH rack +  - Store these aliquots at -15° C to -20° C. 
-Check IQC results, in case outliers call residents.+  - Prior to use, make sure that aliquots should be at room temperature for at least 15 min.
  
-Principle of the procedure used for examinations: +**Procedure to run IQC** 
-The glycerol is phosphorylated by adenosinetriphosphate (ATP) with glycerol kinase (GK) to produce glycerol-3-phosphate and adenosine diphosphate (ADP). Glycerol-3-phosphateis oxidized to dihydroxyacetone phosphate (DAP) by glycerol phosphate oxidase (GPO) producing hydrogen peroxide (H2O2). In a color reaction catalyzed by peroxidase, the H2O2 reacts with 4-aminoantipyrine (4-AAP) and 4-chlorophenol (4-CP) to produce a red colored dye. The absorbance of this dye is proportional to the concentration of triglyceride present in the sample.+  - Press Control order  
 +  - Select Assay /Panel, to be run. 
 +  - Select the control/s and its level/s 
 +  - Give Carrier Number and Position number 
 +  - Press F3 / Add order 
 +  - Put respected carrier in RSH rack 
 +  - Check IQC results, in case outliers call residents.
  
- Sample Preparation: +**10.Principle of the procedure used for examinations:** 
- ● Required SampleVolume: 150 µl of the sample  +Triglycerides are enzymatically hydrolyzed by lipase to free fatty acids and glycerol. The glycerol is phosphorylated by adenosine triphosphate (ATP) with glycerol kinase (GK) to produce glycerol-3-phosphate and adenosine diphosphate (ADP). Glycerol-3-phosphate is oxidized to dihydroxyacetone phosphate (DAP) by glycerophosphate oxidase (GPO) producing hydrogen peroxide (H2O2). In a color reaction catalyzed by peroxidase, the H2O2 reacts with4-aminoantipyrine (4-AAP) and 4-chlorophenol (4-CP) to produce a red colored dye. The absorbance of this dye is proportional to the concentration of triglyceride present in the sample. This analytical methodology is based on the reaction sequence described by Fossati et al.4 and by McGowan et al.5 In this reagent,4-chlorophenol is used rather than 2-hydroxy-3,5-dichloro benzene sulfonate, used in the Fossati and McGowan studies.
- ● Temperature: 37°  +
-Take 150-200µl of the sample from Primary tube to the secondary aliquot. Write the sample ID on the aliquot.  +
-●Procedure to run Patient sample +
-Press Patient order  +
-Select Assay /Panel, to be run. +
-Give Carrier Number and Position number +
-Press F3 / Add order +
-Put respected carrier in RSH rack+
  
-Performance Characteristics:  +**11.Sample Preparation:** 
-● Linearity: up to 1420 mg/dL +  * Required SampleVolume: 150 µl of the sample  
-● The limit of detection (LOD): 5.0 mg/dL +  * Temperature: 37°  
-● The limit of quantification(LOQ): 6.2 mg/dL +  * Take 150-200µl of the sample from Primary tube to the secondary aliquot. Write the sample ID on the aliquot. 
-● Unit:  mg/dL+
  
 +**Procedure to run Patient sample**
 +  - Press Patient order 
 +  - Select Assay /Panel, to be run.
 +  - Give Carrier Number and Position number
 +  - Press F3 / Add order
 +  - Put respected carrier in RSH rack
  
 +**12.Performance Characteristics**: 
 +  - Linearity: up to 1420 mg/dL
 +  - The limit of detection (LOD): 0.62 mg/dL
 +  - The limit of quantification(LOQ): 6.2 mg/dL
 +  - Unit:  mg/dL
  
- Normal and critical ranges:+**Normal and critical ranges:**
  
-Parameter +**Triglyceride** 
-Desirable / Optimal +^Desirable^ Borderline High^High^ 
-Near/ Above optimal +|<200|200-399|400 and above|
-Borderline +
-High +
-Very High+
  
  
-Triglyceride +**13.Laboratory Clinical interpretation:** 
-< 150 mg/dL+The common causes of hyperlipidemia is nephrosis,DM,Endocrine disturbances.
  
 +**14.Interference and cross reaction:**
  
-150-199 mg/dL +InterferingSubstance 
-200-499 mg/dL +  * No interference from Bilirubin up to7.5 mg/dL  
->/= 400 mg/dL+  * No interference from Hemoglobin up to750 mg/dL 
 +  * No interference from Ascorbate up to 1.5 mg/dl  
 +  * No interference from Acetamenophen up to200mg/dl.  
 +  * No interference from Dipyrone up to100mg/dl.  
 +  * No interference from N acetyl L cystenine up to800 mg/dl
  
 +**15.Potential source of variation:** 
 +  * Turn around time (TAT):
 +  * Routine: 6.0 hours
 +  * Urgent: 2.0 hours
  
 +**16.Recording of observation:**
 +  * Software backup 
 +  * Machine raw data
 +**17.torage & Disposal of waste:** Follow storage & discard procedure
  
- Laboratory Clinical interpretation: +**18.Environmental & Safety control:**
-Measurement of triglyceride is important in the diagnosis and management of  hyperlipidemia. These diseases can be genetic or secondary to other disorders including nephrosis, diabetes mellitus, and endocrine disturbances.+
  
- Interference and cross reaction:+**Reagent R1** contains sodium azide.contact with acids liberates very toxic gas
  
-No interference from Bilirubin up to7.5 mg/dL  +**19.Precautions:** 
-No interference from Hemoglobin up to750 mg/dL +  - Wear protective gloves / protective clothing / eye protection 
-No interference from Ascorbate up to 1.5 mg/dl  +  - Do not breathe mist / vapors / spray 
-No interference from Acetamenophen up to200mg/dl.  +  - Wash hands thoroughly after handling 
-No interference from Dipyrone up to100mg/dl.  +  - Keep only in original container
-No interference from N acetyl L cystenine up to800 mg/dl +
- +
-Potential source of variation:  +
-Turn around time (TAT): +
-Routine: 6.0 hours +
-Urgent: 2.0 hours +
- +
- Recording of observation: +
-Software backup  +
-Machine raw data +
-Storage & Disposal of waste: Follow storage & discard procedure +
-Environmental & Safety control: +
-Reagent R1 contains sodium azide.contact with acids liberates very toxic gas +
-Precautions: +
-Wear protective gloves / protective clothing / eye protection +
-Do not breathe mist / vapors / spray +
-Wash hands thoroughly after handling +
-Keep only in original container+
 . .
-References: +**20.References:** 
- +  - US department of labor,Occupational safety and health administration. 
-US department of labor,Occupational safety and health administration. +  - US department of health and human services. Biosafety in MIcrobiological and biomedical laboratories. 
-US department of health and human services. Biosafety in MIcrobiological and biomedical laboratories. +  - World health organization. Biosafety manual,3rd edition. 
-World health organization. Biosafety manual,3rd edition. +  - World health organization. Biosafety manual,3rd edition. 
-World health organization. Biosafety manual,3rd edition. +  - Clinical and laboratory std institute . Protection of Laboratory workers from oocupationally acquired infection; Approved guideline -4th edition. 
-Clinical and laboratory std institute . Protection of Laboratory workers from oocupationally acquired infection; Approved guideline -4th edition. +  - Burtis CA,Ashwood ER,editors,Tietz Textbook of clinical chemistry
-Burtis CA,Ashwood ER,editors,Tietz Textbook of clinical chemistry +
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 +|**Printed copy of this document is considered uncontrolled.** It should be compared with controlled electronic copy before use|
  
 +^ Name of Laboratory : Laboratory Services Sir T. Hospital (LSSTH),Bhavnagar ^^^^
 +^Document No.1^**Document Name**: Triglyceride Examination Procedure^**Unique ID**:LSSTH /BIOCHEM/ SOP-5^^
 +^Issue No. : 01^Issue Date :30/04/2024^Page No.^^
 +^Amend No.^ Amend Date ^Prepared by: Section Incharge^Approved & Issued by: HOD,Biochemistry^