Unisule Pvt. Ltd.

M-15, Industrial Area, Sonepat-131001

CALIBRATION RECORD OF
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
NAME OF INSTRUMENT:
INSTRUMENT CODE
:
UV DETECTOR
:
PUMP
:
Date of Calibration
:
Next Due Date
:
S.NO. WAVE LENGTH PEAK NAME
1
266 nm
Caffeine
2
267 nm
Caffeine
3
268 nm
Caffeine
4
269 nm
Caffeine
5
270 nm
Caffeine
6
271 nm
Caffeine
7
272 nm
Caffeine
8
273 nm
Caffeine
9
274 nm
Caffeine
10
275 nm
Caffeine
11
276 nm
Caffeine

HPLC (SHIMADZU)
US-QC-001
MODEL SPD-10A
MODEL LC-10AT
01-10-2008
01-04-2009
AREA
REMARKS
546.3218
Minimum
558.6945
Area Increasing
586.6356
Area Increasing
597.8326
Area Increasing
612.4568
Area Increasing
649.2658
Area Increasing
659.2357
Maximum
629.3645
Area Decreasing
589.5648
Area Decreasing
574.3956
Area Decreasing
560.3265
Area Decreasing

CONCLUSION:- The UV-DETECTOR of HPLC calibrated over column C-18

using Caffeine as standard in the range of wavelength lying from 266 nm to 276
nm and found that the Detector showed maxima at 272 nm which fall in the
stipulated range of 271 to 274 nm.

ANALYST

Q.C. INCHARGE

CALIBRATION CHART OF HPLC-PUMP

Date of Calibration:- 01-10-2008
Reference
Liquid used

Flow rate
set

Estimated
volume

Weight of calibrated
measuring cylinder

Water

1 ml/min

5 ml

20.4268 gm

Weight of
water +
cylinder
25.4088 gm

Net volume
delivered

Tolerance
+ 5%

4.992 ml

4.75ml to
5.25 ml
9.50ml to
10.5 ml
14.25ml to
15.75 ml

Water

1 ml/min

10 ml

20.4268 gm

30.3859 gm

9.98 ml

Water

1 ml/min

15 ml

35.6132 gm

50.5689 gm

14.98 ml

Reference
Liquid used

Flow rate
set

Estimated
volume

Weight of calibrated
measuring cylinder

Water

2 ml/min

10 ml

20.4268 gm

Weight of
water +
cylinder
30.4016 gm

Water

2 ml/min

20 ml

50.5646 gm

70.4767 gm

19.95 ml

Water

2 ml/min

30 ml

75.4235 gm

105.3349 gm

29.97 ml

Net volume
delivered

Tolerance
+ 5%

9.995 ml

9.50ml to
10.50 ml
19.00ml to
21.00ml
28.50 ml to
31.50 ml

Assuming the weight of water 0.99802 gm/ ml at 20 o C.

CONCLUSION:- The above observation of flow rate is within the limits + limits of inhouse
specification, hence the Calibration of pump is OK in best of our knowledge.

ANALYST

Q.C. INCHARGE