BIOLINE DIAGNOSTICS LLP

H, 1478 DSIDC Industrial area, Narela, Delhi, 110040

STANDARD OPERATING PROCEDURE
Doc. No. QC/STP/005/009 1 /3
Doc. Name STP for CHOLESTEROL (IP) Revision No.

Effective Date Next Review

1.0 Objective:
To the describe procedure for In-process testing of Cholesterol.

2.0 SCOPE
This STP is applicable for the In process testing of Cholesterol.

3.0[2.0] RESPONSIBILITY
Officer/ Executive - Quality Control

4.0[3.0] ACCOUNTABILITY
Manager - Quality Assurance

5.0[4.0] REFRENCE
QA/SOP/001: Writing of Document/Procedure
QC/SOP/022: SOP for STP
6.0[5.0] PROCEDURE
6.1 Visual Check:
6.1.1 Reagent color: Take 1 ml of Cholesterol reagent R1, standard 200 micro liter in a clear
glass test tube and examine under bright light for any turbidity or suspended particles.
R1: Should be clear transparent colorless solution generates foam on shaking.
Standard: Should be clear colorless solution generates foam on shaking.
6.2 Testing procedure: Before performing the test, bring the reagents R1 and Standard at
room temperature. Testing is to be done with biochemistry analyzer.
6.3.1 Set Parameter: Set the below parameter in biochemistry analyzer before carrying out the
test:
Name of Test Cholesterol
Mode- End point

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 BIOLINE DIAGNOSTICS LLP
H, 1478 DSIDC Industrial area, Narela, Delhi, 110040
STANDARD OPERATING PROCEDURE
Doc. No. QC/STP/005/009 2 /3
Doc. Name STP for CHOLESTEROL (IP) Revision No.

Effective Date Next Review

Filter 505 nm
Reaction direction Increasing
Temperature 37⁰C
Incubation time 5 min
Ref value 200-239 mg/dl
High Linearity 1000 mg/dl
Standard conc. 200 mg/dl
6.2.2 Assay Procedure:
6.2.2.1 Standard assay: Carry out the standard testing as per below table.
6.2.2.2 Table
Assay Blank Approved Standard/ Reference Under Test
Calibrator (4 test tubes) Standard (4 test tube)
Reagent R1 1ml 1ml 1ml
Standard NA 10µl 10µl
6.2.2.3 Mix all the tubes and incubate for 5 min at 37⁰C. After completion of incubation time
read the absorbance of Approved Standard/Reference Calibrator and Under Test
Standard against the Reagent blank. Reagent blank should not be more than 0. 050 OD.
Calculate the Average absorbance of Approved Std/Ref Calibrator tubes and Average
absorbance of Under test tubes. Concentration of Under test Standard is calculated using
the below formula

Conc of U/T STD = Abs of U/T STD /Ref Cal- Abs of Rgt blank X Concn of approved Std/Ref Cal
Abs of App STD /Ref Cal- Abs of Rgt blank

The concentration of Under test standard should fall within the acceptable limit of the
standard concentration (199-201).

6.2.2.2 Validation of standard using Quality control/Sample and Linearity samples

Once the Under test standard is Approved against previously QC passed standard/Reference
calibrator we need to validate it against third part control or reference lab samples as below
table.

6.2.2.3 Assay table

Prepared by: Reviewed by: Approved by:
 BIOLINE DIAGNOSTICS LLP
H, 1478 DSIDC Industrial area, Narela, Delhi, 110040
STANDARD OPERATING PROCEDURE
Doc. No. QC/STP/005/009 3 /3
Doc. Name STP for CHOLESTEROL (IP) Revision No.

Effective Date Next Review

Reagent Blank Standard Control/Sample High Linearity
Reagent R1 1ml 1ml 1ml 1ml
Standard NA 10µl NA NA
Control/ Sample NA NA 10 µl NA
High Linearity N/A NA NA As per need

6.2.3 Read the absorbance of Standard, Control/Sample, and High linearity tubes against the
Reagent blank tube after 5 min of incubation at 37⁰C, the reagent should not be more than
0. 050 OD. There should not be any significant increase in the absorbance of the reagent.
6.2.5 The result of Control, High linearity obtained after the completion of the test should fall
within the acceptable limit of the Control values printed in the Value sheet of control
being used e.g. (Randox 2, Randox 3, Biorad 1, Biorad 2 etc….) and high linearity
should also be within acceptable range.
6.3 Prepare Analysis report
6.3.1 After completion of the test, In-process testing analysis report should be prepared by the
responsible person and released IP reagent as per In process SOP (QC/SOP/005).

7.0 RECORDS/ FORMAT

IP TEST REPPORT FORMATS-QC/IHS/005//009

8.0 ABBREVIATIONS

SOP - Standard Operating Procedure

STP-Standard Testing Procedure
IP - In process

9.0 CHANGE HISTORY DETAILS

Change control no. Change details Superseded Document No. and revision status

New initiation N/A N/A

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