1.0 OBJECTIVE:
To
lay down the operating procedure of High Performance Liquid Chromatography (HPLC).
2.0 SCOPE:
This
shall be applicable to the High
Performance Liquid Chromatography (HPLC) of Quality Control Instrument Laboratory.
3.0 RESPONSIBILITY:
3.1
Technical
Assistant/ Helper.
3.2
Q.C Analyst.
4.0 ACCOUNTABILITY:
4.1
Manager-Quality
Control.
5.0 MATERIAL AND
EQUIPMENT:
5.1
High Performance Liquid Chromatography (HPLC).
5.2
Sample.
6.0 PROCEDURE:
6.1
Operation:
6.1.1
Power
ON the system:
6.1.1.1
Power on the pump, detector, and any other component like computer.
6.1.2
Purging
the pump:
6.1.2.1
Ensure that the solvent inlet filter, inside the solvent bottle, is in
vertical position. Pump must be purged with every new solvent.
6.1.3
Set
the analytical parameters:
6.1.3.1
After power ON the status menu will appear to change the parameters.
6.1.3.2
Change flow rate and wave length according to requirement.
6.1.3.3
Run purified water approximately 20 minutes.
6.1.3.4
After that run mobile phase for approximately 30 minutes to saturate
column with mobile phase.
6.1.3.5
Inject standard or sample only when reading on detector is zero.
6.1.3.6
Press auto zero before each injection.
6.1.4
Shut
down:
6.1.4.1
After completion
of work following steps must be taken to save the life of system.
6.1.4.2
Wash with 80ml of
water (HPLC Grade) as under.
6.1.4.3
Dip inlet tube in
the water container and press [RUN]. The water will go through the pump,
Injector, column, and detector. In this way all salts and inorganic materials
will be washed. After 70 ml of water passing press [STOP] key.
6.1.4.4
Finally pass 50 ml
(approximately) methanol through the column to save the column. After 50 ml of
methanol passing press [STOP] key.
6.1.4.5
Power OFF the
pump, Detector.
6.2
Precautions:
6.2.1
Check all tubing, wiring and column that should not be blocked broken
or pricked.
6.2.2
Column must be compatible to the analytical method as well as mobile
phase. Make corrections if necessary.
6.2.3
The availability of reference samples, solvents and chemicals must be
confirmed.
6.2.4
Table having instrument should be free of vibration.
7.0 REVISION LOG:
|
Revision No.
|
Effective Date
|
Reason
|
|
00
|
|
New
SOP
|
8.0 REFERENCES:
8.1
Not Applicable.
9.0 ANNEXURES:
9.1
Not Applicable.
10.0
ABBREVIATIONS:
|
Abbreviation
|
Expanded Form
|
|
SOP
|
Standard operating
procedure
|
|
&
|
And
|
|
No.
|
Number
|
|
Ltd.
|
Limited
|
|
Q.C
|
Quality control
|
|
QCEO
|
Quality control
equipment operation
|
|
F
|
Format
|
|
ml
|
Milliliter
|
|
HPLC
|
High Performance Liquid Chromatography
|
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