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Document Title : HPLC CALIBRATION
Reference SOP : PROCEDURE FOR OPERATION&CALIBRATION OF
HPLC
Reference SOP No. : PB/SOP/QCD/001/01 Format No. :
F/PB/QCD/001/01
Annexure No. : III Page No. : 1 of 1
HPLC ID No: ------------------------------- Column ID No :
-------------------------
Make : ------------------------------ Model :
--------------------------
Date of calibration: ------------------------ Calibration due
Date: ---------------------------
Frequency : Half Yearly
1. FLOW RATE:
Weight per ml of water
Wt of Empty Pycnometer g
Wt / ml
=g/ml
Wt of Filled Pycnometer g
Wt of Water g 25.0
Set flow
Wt. of
empty
Cylender
(W1)
Wt. of
Cylender +
Water after
10 minutes
(W2)
Flow rate
(ml/min)=
[(W2-W1)/10 xwt/ml]
Average
flow Rate
(ml/min)
Limit + 2.0%
Minimum
(ml/min)
Maximu
m
(ml/min)
1.0 ml/min
g g
0.98 1.02g g
g g
2.0 ml/min g g 1.96 2.04
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Document Title : HPLC CALIBRATION
Reference SOP : PROCEDURE FOR OPERATION&CALIBRATION OF
HPLC
Reference SOP No. : PB/SOP/QCD/001/01 Format No. :
F/PB/QCD/001/01
Annexure No. : III Page No. : 2 of 1
2. Linearity (Detector) :
Reagent Name : --------------------------------------- Reagent
B.No: -----------------------------
Mobile phase :70:30Methanol&Water Balance
ID.No:------------------------------
Column : ----------------------------------- Column ID
.No:-----------------------------
Wave length : ---------------------------------- Flow Rate
(1ml/Min) :---------------------
Sample Solution A: Weigh accurately about ____(100 mg) of
Caffeine Anhydrous Pure in 100 ml of
Volumetric Flask, dissolve and make up to mark with Mobile
Phase.
Sample Solution B: dilute 1 ml of this solution A to 100 ml with
Mobile Phase (10 ppm).
Sample Solution C:dilute 2 ml of this solution B to 100 ml with
Mobile Phase (20 ppm).
Sample Solution D: dilute 3 ml of this solution C to 100 ml with
Mobile Phase (30 ppm).
Sample Solution E: D dilute 4 ml of this solution D to 100 ml
with Mobile Phase (40 ppm).
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Document Title : HPLC CALIBRATION
Reference SOP : PROCEDURE FOR OPERATION&CALIBRATION OF
HPLC
Reference SOP No. : PB/SOP/QCD/001/01 Format No. :
F/PB/QCD/001/01
Annexure No. : III Page No. : 3 of 1
S.NoConcentrati
onArea Mean Area
Correlation
Coefficient (r)
Acceptance
Criteria
01 10 ppm
0.98 to
1.000
02 20 ppm
03 30ppm
04 40ppm
05 50ppm
Calibrated by: ------------------- Checked by:
--------------------
Date : ----------------- Date : -------------------
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8/11/2019 HPLC Calibration Format
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Document Title : HPLC CALIBRATION
Reference SOP : PROCEDURE FOR OPERATION&CALIBRATION OF
HPLC
Reference SOP No. : PB/SOP/QCD/001/01 Format No. :
F/PB/QCD/001/01
Annexure No. : III Page No. : 5 of 1
4. Wavelenght Accuracy:
Sample Solution A : Weigh accurately about ____(100 mg) of
Caffeine Anhydrous Pure in 100 ml of
Volumetric Flask, dissolve and make up to mark with Mobile
Phase.
Sample Solution B: Further dilute 2 ml of this solution to 100
ml with Mobile Phase (20 g/ml).
S.No Wavelength (nm) Area Maxima at Acceptance Criteria
1 270
273nm
2 271
3 272
4 273
5 274
6 275
7 276
Instrument Calibration meets the requirement /doesn`t meets the
requirements
Calibrated by: ------------------- Checked by:
--------------------
Date : ----------------- Date : -------------------
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8/11/2019 HPLC Calibration Format
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Document Title : HPLC CALIBRATION
Reference SOP : PROCEDURE FOR OPERATION&CALIBRATION OF
HPLC
Reference SOP No. : PB/SOP/QCD/001/01 Format No. :
F/PB/QCD/001/01
Annexure No. : III Page No. : 6 of 1
