Differences

This shows you the differences between two versions of the page.

Link to this comparison view

Both sides previous revision Previous revision
triglyceride [2025/01/28 04:30] – admintriglyceride [2025/01/28 04:31] (current) – admin
Line 1: Line 1:
-' +|[[clinical_biochemistry_section|Home]]|[[clinical_biochemistry|]]|[[examination_procedures|]]|
-Purpose of examination:  +
-●Triglyceride estimation from serum or plasma by Glycerol Phosphate Oxidase method.+
  
-Responsibility and Authority:  +                      Triglyceride Examination Procedure 
-● Calibration: Technician  +**1.Purpose of examination:**  
-● Quality Control: Technician  +Triglyceride estimation from serum or plasma by Glycerol Phosphate Oxidase.
-● Routine operation: Technician  +
-● Overall Monitoring: Quality Manager+
  
- Sample Details:  +**2.Responsibility and Authority:**  
-● Type of Sample: Serum,  Plasma +  * Calibration: Technician  
-● Type of container and additives: Plain without any additives  +  * Quality Control: Technician  
-● Patient Preparation: As per Primary Sample Collection Manual {{Sample collection manual}} +  * Routine operation: Technician  
-● Stability: At Room temperature 18°–28°C (64°–82°F) stability ≤ 24 hours +  * Overall Monitoring: Quality Manager
-● Handling and transport: As per Primary Sample collection manual  +
-● Storage: 24 hours at 2-8° C+
  
-Required Equipment:  +**3.Sample Details:**  
-●Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer+  * Type of Sample: Serum,  Plasma 
 +  * Type of container and additives: Plain without any additives  
 +  * Patient Preparation: As per Primary Sample Collection Manual  [[sample_collection_manual|]] 
 +  * Stability: At Room temperature 18°–28°C (64°–82°F) stability ≤ 24 hours 
 +  * Handling and transport: As per Primary Sample collection manual  
 +  * Storage: 24 hours at 2-8° C
  
-Required reagents: +**4.Required Equipment:**  
-       R1  ATP (2.5 mmol/L),  +Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer
-              Mg2+ (2.5 mmol/L), +
-              4-aminoantipyrine (0.4 mmol/L),  +
-              4-chlorophenol (2 mmol/L), +
-              Peroxidase (horseradish) (> 2000 U/L),  +
-              GK (microbial) (> 600 U/L), +
-              GPO (microbial) (> 6000 U/L), +
-              Lipoprotein lipase (microbial) (> 3000 U/L).+
  
- Reagent Handling +**5.Required reagents:** 
- ● 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 +  * R1 84ml Active ingredients: ATP (2.5 mmol/L), Mg2+ (2.5 mmol/L), 
-Reagent Storage and stability +  * 4-aminoantipyrine (0.4 mmol/L), 4-chlorophenol (2 mmol/L), 
-Unopened reagent stable at 2-8°C until expiration date.+  * Peroxidase (horseradish) (> 2000 U/L), GK (microbial) (> 600 U/L), 
 +  * GPO (microbial) (> 6000 U/L), Lipoprotein lipase (microbial) (> 3000 U/L).
  
-On board System temperature reagent is  stable for 42 days +**6.Reagent Handling** 
-Instability or deterioration should be suspected if there are precipitates, visible signs of leakage or +  * 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 
-contamination, turbidity, or if calibration or controls do not meet the appropriate criteria.+  * Reagent Storage and stability 
 +  * 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.
  
-Calibration Procedure: 
-● Consolidated Chemistry Calibrator 
  
-● Frequency: 
-Reagent lot change 
-QC out of range 
-After service or maintenance 
-Replacement in any parts of Instrument 
-Procedure: 
-Start the equipment.WDI abbotte fully.docx 
-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: +**7.Calibration Procedure:** 
-Name: Bio Rad Level 1 & 2 +  * Consolidated Chemistry Calibrator 
-Frequency: As per Quality Control Procedure +  * Frequency: 
-●Procedure for Reconstitution of IQC  +  * Reagent lot change 
-Reconstitution of QC material with 5 ml Distilled water by using calibrated fixed volume pipette. +  * QC out of range 
-Leave to stand for 30 min in the dark place. +  * After service or maintenance 
-Swirl gently several times during the reconstitution period to ensure that the contents are completely dissolved. +  * Replacement in any parts of Instrument
-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. +
-Prepare aliquots of 150 µl from the reconstituted QC material. +
-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.+
  
 +**8.Procedure:**
 +  - Start the equipment.WDI abbotte fully.docx
 +  - 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.
  
-●Procedure to run IQC +**9.Quality control Procedure:** 
-Press Control order  +  * ** Name:** Bio Rad Level 1 & 2 
-Select Assay /Panel, to be run. +  * **Frequency:** As per Quality Control Procedure
-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.+
  
-Principle of the procedure used for examinations: +**Procedure for Reconstitution of 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.+  - Reconstitution of QC material with 5 ml Distilled water by using calibrated fixed volume pipette. 
 +  - Leave to stand for 30 min in the dark place. 
 +  - Swirl gently several times during the reconstitution period to ensure that the contents are completely dissolved. 
 +  - 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. 
 +  - Prepare aliquots of 150 µl from the reconstituted QC material. 
 +  - 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.
  
- Sample Preparation: +**Procedure to run IQC** 
- ● Required SampleVolume: 150 µl of the sample  +  - Press Control order  
- ● Temperature: 37°  +  - Select Assay /Panel, to be run. 
-Take 150-200µl of the sample from Primary tube to the secondary aliquot. Write the sample ID on the aliquot.  +  - Select the control/s and its level/s 
-●Procedure to run Patient sample +  - Give Carrier Number and Position number 
-Press Patient order  +  - Press F3 / Add order 
-Select Assay /Panel, to be run. +  - Put respected carrier in RSH rack 
-Give Carrier Number and Position number +  - Check IQC results, in case outliers call residents.
-Press F3 / Add order +
-Put respected carrier in RSH rack+
  
-Performance Characteristics:  +**10.Principle of the procedure used for examinations:** 
-● Linearity: up to 1420 mg/dL +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.
-● The limit of detection (LOD): 5.0 mg/dL +
-● The limit of quantification(LOQ): 6.2 mg/dL +
-● Unit:  mg/dL+
  
 +**11.Sample Preparation:**
 +  * Required SampleVolume: 150 µl of the sample 
 +  * 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
  
- Normal and critical ranges:+**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
  
-Parameter +**Normal and critical ranges:**
-Desirable / Optimal +
-Near/ Above optimal +
-Borderline +
-High +
-Very High+
  
 +**Triglyceride**
 +^Desirable^ Borderline High^High^
 +|<200|200-399|400 and above|
  
-Triglyceride 
-< 150 mg/dL 
  
 +**13.Laboratory Clinical interpretation:**
 +The common causes of hyperlipidemia is nephrosis,DM,Endocrine disturbances.
  
-150-199 mg/dL +**14.Interference and cross reaction:**
-200-499 mg/dL +
->/= 400 mg/dL+
  
 +InterferingSubstance
 +  * No interference from Bilirubin up to7.5 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
  
- Laboratory Clinical interpretation: +**16.Recording of observation:** 
-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.+  * Software backup  
 +  * Machine raw data 
 +**17.torage & Disposal of waste:** Follow storage & discard procedure
  
- Interference and cross reaction:+**18.Environmental & Safety control:**
  
-No interference from Bilirubin up to7.5 mg/dL  +**Reagent R1** contains sodium azide.contact with acids liberates very toxic gas
-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+
  
-Potential source of variation:  +**19.Precautions:** 
-Turn around time (TAT): +  - Wear protective gloves / protective clothing / eye protection 
-Routine: 6.0 hours +  - Do not breathe mist / vapors / spray 
-Urgent: 2.0 hours +  - Wash hands thoroughly after handling 
- +  - Keep only in original container
- 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 +
- +
- +
- +
- +
- +
  
  
 +|**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^