Internal Calibration Check Issue No.

: 01
AVITECH
Nutrition (P) Ltd. HPLC SYSTEM Issue Date: 07-09-2018
Revision No.: 01
(Analytical Division) (Agilent Prime LC 1260 Infinity II)

Revision Date: 08-03-2019

Format No. ANT/FMT/ICC /13 Page No: 1/5

Instrument ID. No.: ANT/T-EQP/62 Date of calibration:

Frequency of calibration: Quarterly Next due date for calibration:

(l) FLOW RATE ACCURACY TEST:

Mobile Phase: Water (HPLC Grade,100%) Stop Watch ID. No: ………….
Stop Watch Calibration Due on:

Sr. No. Set flow rate Theoretical Actual Actual flow Limit
Seconds seconds(Y) rate
1 1.0 ml/min 600 second 0.990 to1.010
2 1.5 ml/min 400 second 1.485 to 1.515
3 2.0 ml/min 300 second 1.980 to 2.020

Equation: 10 X 60
Y
(2) RETENTION TIME REPRODUCIBILITY: For DAD
Take …………….mg of Caffeine and dilute to …………. ml with mobile phase. Further dilute 1 ml to
100 ml with mobile phase (final concentration 10ppm).

Injection No. Retention time Limit

1
2
3 % RSD not more than 1.0
4
5
Mean

Calibrated By Checked By
Analyst/Sr. Analyst Sr. Manager (QC)
SD
% RSD

Internal Calibration Check Issue No.: 01

AVITECH
Nutrition (P) Ltd. HPLC SYSTEM Issue Date: 07-09-2018
Revision No.: 01
(Analytical Division) (Agilent Prime LC 1260 Infinity II)

Revision Date 08-03-2019

Format No. ANT/FMT/ICC /13 Page No: 2/ 5
(3) INJECTION PRECISION: For DAD

Injection No. Area Limit

1
2
3 % RSD not more than 2.0
4
5
Mean
SD
% RSD

(4) INJECTOR LINEARITY: For DAD

Injection No. Injection Volume Area Limit

1
2
3
4
5
Correlation coefficient NLT 0.990

(5) DETECTOR LINEARITY: For DAD

Injection No. Injection Volume Area Limit

Calibrated By Checked By
Analyst/Sr. Analyst Sr. Manager (QC)
1
2
3
4
5
Correlation coefficient NLT 0.990

Internal Calibration Check Issue No.: 01

AVITECH
Nutrition (P) Ltd. HPLC SYSTEM Issue Date: 07-09-2018
Revision No.: 01
(Analytical Division) (Agilent Prime LC 1260 Infinity II)

Revision Date: 08-03-2019

Format No. ANT/FMT/ICC /13 Page No: 3/5
(6) CARRY OVER TEST: For DAD

Sr. No. Mean area of solution Area of caffeine in % Carry Over Limit
blank injection NMT 0.20%

(7) WAVE LENGTH ACCURACY:

Wave 265 266 267 268 269 270 271 272 273
length nm.
Absorbance
Limit LIMIT 272 ± 2 nm
Wave length 274 275
nm
Absorbance
Limit

(8) GRADIENT COMPOSITION:

Composition Using A & B Composition Using C & D Limit

Composition

Calibrated By Checked By
Analyst/Sr. Analyst Sr. Manager (QC)
 m.AU % of m.AU % of
composition composition
C100

79.0 to 81 %
C80
59 to 61 %
C60
39.0to 41.0 %
C40
19.0to 21.0 %
C20

Internal Calibration Check Issue No.: 01

AVITECH
Nutrition (P) Ltd. HPLC SYSTEM Issue Date: 07-09-2018
Revision No.: 01
(Analytical Division) (Agilent Prime LC 1260 Infinity II)

Revision Date: 08-03-2019

Format No. ANT/FMT/ICC /13 Page No: 4/5
Calculation: for channel A & B

Actual concentration of 20 % (B) = ---------------- x100

Actual concentration of 40 % (B) = ---------------- x100

Actual concentration of 60 % (B) = ---------------- x100

Actual concentration of 80 % (B) = ---------------- x100

Calculation: for channel C& D

Actual concentration of 20 % (D) = ---------------- x100

Actual concentration of 40 % (D) = ---------------- x100

Actual concentration of 60 % (D) = ---------------- x100

Actual concentration of 80 % (D) = ---------------- x100

Calibrated By Checked By
Analyst/Sr. Analyst Sr. Manager (QC)
(9) RETENTION TIME REPRODUCIBILITY: For FLD
Take ……………..mg of Anthracene and dilute to …………….ml with 100% Acetonitrile. Further
dilute 1 ml to 100 ml with mobile phase.

Injection No. Retention time Limit

1
2
3 % RSD not more than 1.0
4
5
Mean
SD
% RSD

Internal Calibration Check Issue No.: 01

AVITECH
Nutrition (P) Ltd. HPLC SYSTEM Issue Date: 07-09-2018
Revision No.: 01
(Analytical Division) (Agilent Prime LC 1260 Infinity II)

Revision Date: 08-03-2019

Format No. ANT/FMT/ICC /13 Page No: 5/5
(10) INJECTION PRECISION: For FLD

Injection No. Area Limit

1
2
3 % RSD not more than 2.0
4
5
Mean
SD

Calibrated By Checked By
Analyst/Sr. Analyst Sr. Manager (QC)
% RSD

Checking of Column Compartment:

(11) COLUMN OVEN TEMPERATURE: Thermometer ID No.

Sr. No Set Temperature Displayed Temp. Observed temp of Limits:

thermometer
1 25° C 24 to 26 ° C
2 40 ° C 39 to 41 ° C

3 60 ° C 59 to 61 ° C

(12) SAMPLER COOLER TEMPERATURE: Thermometer ID No.

Sr. No Set Temperature Observed temp of Limits:

thermometer
1 10° C +/ - 1.0° C
of set temperature
2 40 ° C

Remark: Instrument meets/does not meet with acceptance criteria. The calibration is OK / Not OK.

Calibrated By Checked By
Analyst/Sr. Analyst Sr. Manager (QC)