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+|[[clinical_biochemistry_section|Home]]|[[clinical_biochemistry|]]|[[examination_procedures|]]|
+                      HDL Cholesterol Examination Procedure
+**1.Purpose of examination:**
+HDL Cholesterol estimation from serum or plasma by Accelerator Selective Detergent method.
-Purpose of examination:
+**2.Responsibility and Authority:**
-●Triglyceride estimation from serum or plasma by Accelerator Selective Detergent method.
+  * Calibration: Technician
+  * Quality Control: Technician
+  * Routine operation: Technician
+  * Overall Monitoring: Quality Manager
-Responsibility and Authority:
+**3.Sample Details:**
-● Calibration: Technician
+  * Type of Sample: Serum,  Plasma
-● Quality Control: Technician
+  * Type of container and additives: Plain without any additives
-● Routine operation: Technician
+  * Patient Preparation: As per Primary Sample Collection Manual [[sample_collection_manual|]]
-● Overall Monitoring: Quality Manager
+  * 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
- Sample Details:
+**4.Required Equipment:**
-● Type of Sample: Serum,  Plasma
+Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer
-● 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 Equipment:
+**5.Required reagents:**
-●Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer
-Required reagents:
 R1:  Cholesterol oxidase (E. coli) (< 1000 U/L),
-        Peroxidase (Horseradish) (< 1300 ppg U/L),
+Peroxidase (Horseradish) (< 1300 ppg U/L),
-        N, N-bis (4-sulfobutyl)-m-toluidine-disodium (DSBmT) (< 1.0 mmol/L),
+N, N-bis (4-sulfobutyl)-m-toluidine-disodium (DSBmT) (< 1.0 mmol/L),
-        Accelerator (< 1.0 mmol/L),
+Accelerator (< 1.0 mmol/L),
-        Ascorbic oxidase (Cucurbita sp.) (< 3000 U/L).
+Ascorbic oxidase (Cucurbita sp.) (< 3000 U/L).
- 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
-Reagent Storage and stability
-Unopened reagent stable at 2-8°C until expiration date.
-On board System temperature reagent is  stable for 28 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:
+**6.Reagent Handling**
-● Consolidated Chemistry Calibrator
+  * 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
+  * Unopened reagent stable at 2-8°C until expiration date.
+  * On board System temperature reagent is  stable for 28 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.
-● Frequency:
+**7.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:
+**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.
+**8.Quality control Procedure:**
+  * Name: Bio Rad Level 1 & 2
+  * Frequency: As per Quality Control Procedure
+**Procedure for Reconstitution of IQC**
+  * 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.
-●Procedure to run IQC
+**Procedure to run IQC**
-Press Control order
+  * Press Control order
-Select Assay /Panel, to be run.
+  * Select Assay /Panel, to be run.
-Select the control/s and its level/s
+  * Select the control/s and its level/s
-Give Carrier Number and Position number
+  * Give Carrier Number and Position number
-Press F3 / Add order
+  * Press F3 / Add order
-Put respected carrier in RSH rack
+  * Put respected carrier in RSH rack
-Check IQC results, in case outliers call residents.
+  * Check IQC results, in case outliers call residents.
-Principle of the procedure used for examinations:
+**9.Principle of the procedure used for examinations:**
-This method is based on accelerating the reaction of cholesterol oxidase (CO) with non-HDL
+  * This method is based on accelerating the reaction of cholesterol oxidase (CO) with non-HDL
 unesterified cholesterol and dissolving HDL cholesterol selectively using a specific detergent. In the first reagent, non-HDL unesterified cholesterol is subject to an enzyme reaction and the peroxide generated is consumed by a peroxidase reaction with DSBmT yielding a colorless product. The second reagent consists of a
 detergent (capable of solubilizing HDL cholesterol), cholesterol esterase (CE), and chromagenic coupler to develop color for the quantitative determination of HDL cholesterol.
- Sample Preparation:
+**10.Sample Preparation:**
- ● Required SampleVolume: 150 µl of the sample
+  * Required SampleVolume: 150 µl of the sample
- ● Temperature: 37°
+  * 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:
+**Procedure to run Patient sample**
-● Linearity: up to 180 mg/dL
+  * Press Patient order
-● The limit of detection (LOD): 5.0 mg/dL
+  * Select Assay /Panel, to be run.
-● The limit of quantification(LOQ): 2.5 mg/dL
+  * Give Carrier Number and Position number
-● Unit:  mg/dL
+  * Press F3 / Add order
+  * Put respected carrier in RSH rack
+**11.Performance Characteristics:**
+  * Linearity: up to 180 mg/dL
+  * The limit of detection (LOD): 5.0 mg/dL
+  * The limit of quantification(LOQ): 2.5 mg/dL
+  * Unit:  mg/dL
+**Normal and critical ranges:**
- Normal and critical ranges:
+^Parameter^Desirable / Optimal^Near/ Above optimal^Borderline^High^Very High^
+|HDL Cholesterol|>60 mg/dL|-|< 40 mg/dL|-|
-Parameter
+**12.Laboratory Clinical interpretation:**
-Desirable / Optimal
+The principle role of HDL cholesterol in lipid metabolism is the uptake and transport of cholesterol from peripheral tissues to the liver through a process known as reverse cholesterol transport proposed cardioprotective mechanism).Low HDL cholesterol levels are strongly associated with an increased risk of coronary heart disease.
-Near/ Above optimal
-Borderline
-High
-Very High
+**13.Interference and cross reaction:**
+  * No interference from Bilirubin up to32.6mg/dL
+  * No interference from Hemoglobin up to1000mg/dL
+  * No interference from Intralipid up to1000 mg/dL
+  * No interference from Ascorbic acid up to 2.9mg/dl
+**14.Potential source of variation:**
+  - Turn around time (TAT):
+  - Routine: 6.0 hours
+  - Urgent: 2.0 hours
-HDL Cholesterol
+**15.Recording of observation**:
->60 mg/dL
+Software backup
+Machine raw data
+**16.Storage & Disposal of waste:** Follow storage & discard procedure
--
+**17.Environmental & Safety control:**
-< 40 mg/dL
+Reagent R1 contains sodium azide.contact with acids liberates very toxic gas
--
+**18.Precautions:**
+  * Wear protective gloves / protective clothing / eye protection
+  * Do not breathe mist / vapors / spray
+  * Wash hands thoroughly after handling
+  * Keep only in original container
- Laboratory Clinical interpretation:
+**19.References:**
-The principle role of HDL cholesterol in lipid metabolism is the uptake and transport of cholesterol from peripheral tissues to the liver through a process known as reverse cholesterol transport proposed cardioprotective mechanism).3 Low HDL cholesterol levels
-are strongly associated with an increased risk of coronary heart disease.
- Interference and cross reaction:
-No interference from Bilirubin up to32.6mg/dL
-No interference from Hemoglobin up to1000mg/dL
-No interference from Intralipid up to1000 mg/dL
-No interference from Ascorbic acid up to 2.9mg/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:
 US department of health and human services. Biosafety in MIcrobiological and biomedical laboratories.
 World health organization. Biosafety manual,3rd edition.
 Gotto AM. Lipoprotein metabolism and etiology of hyperlipidemia.Hosp pract 1988;23
+|**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**: HDL Cholesterol 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^