Category: Calibration

Calibration of Auto Titrator

 

1. CALIBRATION OF pH ELECTRODE

Sr. No.	Standard pH buffers used  at 25oC	Batch No.	Validity
1	Buffer pH 4.01
2	Buffer pH 7.00
3	Buffer pH 9.18

 

Acceptance Criteria:

1. The observed pH of buffer solutions should not be more than ± 0.05.
2. Slope obtained should be between 80% to 120%.

Observations:

 

Buffers used for calibration	Observed pH	Slope obtained
pH 4.01
pH 7.00
pH 9.18

 

1. CALIBRATION OF BURETTE

Sr.No	Volume dispensed by Burette in ml	Specification    ± 1 %	Observation
			Wt. of empty Beaker (W1g)	Wt. of empty Beaker + sample (W2g)	Wt. of sample W3=(W2-W1)	Sample volume = [W3 / 0.99707] ml
1	0.5 ml	0.49 ml- 0.51ml
2	5.0 ml	4.95 ml-5.05ml
3	10.0 ml	9.9 ml- 10.1ml
4	25.0 ml	24.75ml-25.25ml

 D-Density of water is 0.99707 g/ml at 25°C

 

• REPEATABILITY- By Standardization of 0.1 M Sodium Hydroxide Solution

 

1. Preparation- 1:-

Weighed  accurately ___________ g  of  Potassium hydrogen phthalate in  reaction vessels added  _____ ml  of  purified water,  sonicated  for ________ minutes to  dissolve and   titrated with 0.1 M Sodium hydroxide Solution.

2. Preparation- 2:-

Weighed  accurately ___________ g  of  Potassium hydrogen phthalate in  reaction vessels added  _____ ml  of  purified water,  sonicated  for ________ minutes to  dissolve and   titrated with 0.1 M Sodium hydroxide Solution.

3. Preparation- 3:-

Weighed  accurately ___________ g  of  Potassium hydrogen phthalate in  reaction vessels added  _____ ml  of  purified water,  sonicated  for ________ minutes to  dissolve and   titrated with 0.1 M Sodium hydroxide Solution.

Sr. No.	Weight of KHP (gm)	Burette Reading (ml)	Observed Molarity (M)	Mean Molarity (M)	Observed           % RSD
1.
2.
3.

 

KHP= Potassium Hydrogen Phthalate

Acceptance Criteria:

1. The observed Molarity should not be more than ± 0.01M
2. The observed % RSD should not be more than 0.2%

 

Standards/Chemicals Details:

Sr. No.	Standards used	Batch No.	Validity
1	Potassium Hydrogen Phthalate
2	Sodium hydroxide

 

Remark: The result Complies/Does not complies as per acceptance criteria.

 

Performed By: __________                   Checked By: _____

 Date: ____________                                   Date: ______________

 

“End of Document”

CALIBRATION OF VOLUMETRIC GLASSWARE

 

3.2     PROCEDURE FOR CALIBRATION OF VOLUMETRIC FLASKS:

 

3.2.1  Label the volumetric flasks with serial numbers or note down the number, which is present on flask.

3.2.2  Weigh the volumetric to be calibrated on a calibrated analytical balance and note down its weight (w1).

3.2.3  Fill the volumetric flask upto the mark with Distilled water (Which is adjusted to 25 degrees) and close the flask with the stopper immediately.

3.2.3  Weight the volumetric flask again (w2).Calculate the weight of water by deducting (w2- w1).

3.2.4  Measured weight of water is converted into volume by using the following factor: 1ml of water at 25 degrees is equivalent to 0.99602 grams i.e. 0.99602 grams of water is equivalent to 1 ml of water. Factor = 1/0.99602 = 1.004. Record the results in the calibration format and compare with the limits given in the acceptance criteria.

 

3.3     PROCEDURE FOR CALIBRATION OF PIPETTES:

 

3.3.1  Note down the serial numbers of the pipettes to be calibrated.

3.3.2  Weigh an empty clean dried stopper flask or weighing bottle and note down its weight (w1).

3.3.3  Fill the pipette with water up to the mark and adjust to the lower meniscus of the water.

3.3.4  The water drop adhering to the jet is removed by bringing the surface of water contained in a beaker to the jet of the pipette and remove without jerking.

3.3.5  Release the water into weighed beaker and note down its weight w2.

3.3.6  Held the pipette so that the jet of the pipette is in contact with the side of the wall, if necessary incline the pipette slightly.

3.3.7  Stopper the flask immediately and note down its weight (w2).

3.3.8  The weight of water is converted into measured volume by multiplying with the factor (1.004).

3.3.9  Record the results in the calibration format and compare the results with the limits given in the acceptance criteria.

 

3.4     PROCEDURE FOR CALIBRATION OF BURETTE:

 

          3.4.1  Note down the serial number of the Burettes to be calibrated.

          3.4.2  Weigh an empty, dried Beaker and note down its weight (w1).

3.4.3  Fill the Burette with the Purified water upto the mark, taking precautions that no air bubbles should present in the Burette.

3.4.4  Transfer the volume to be calibrated to weighed beaker and note down its weight (w2).

3.4.5  Calculate the measured water weight by deducting w1 from w2 (w2-w1).

3.4.6  The weight of water shall be converted into volume by multiplying with 1.004.

3.4.7  The accuracy of following volumes has to be checked for the burette 10ml, 25ml and 50ml.

3.4.8  Record the results in the calibration format and compare with the limits given in the acceptance criteria.

3.4.9  Before calibrating the Burette, it is essential to establish that the burette is satisfactory with regard to a) leakage b) delivery time.

 

 

 

“End of Document”

 

Calibration of HPLC

  

1. Calibration of Pumps:-

1.1 Flow Accuracy and Flow precision

 Observation table for Accuracy of Flow rate

Sr. No.	Set flow rate in mL/ min	Time require to collect 10mL volume				Observed flow rate in mL/ min	Acceptance criteria
		1	2	3	Average
1	0.5						0.49-0.51
2	1.0						0.98-1.02
3	1.5						1.47-1.53
4	2.0						1.96-2.04
5	5.0						4.90-5.10

 

Observation Table for Precision of flow rate

Sr. No.	Set flow rate in mL/ min	Observed flow rate in mL/min				%RSD	Acceptance criteria
		1	2	3	Average
1	0.5						≤ 2.0%
2	1.0						≤ 2.0%
3	1.5						≤ 2.0%
4	2.0						≤ 2.0%
5	5.0						≤ 2.0%

 

 

• 1.2 Gradient Accuracy:-

  Observation table for Channel A and B

Eluent Composition	Hx	Calculated    Bx (%)	Acceptance Criteria
20%			19.4-20.6
40%			38.8-41.2
60%			58.2-61.8
80%			77.6-82.4
100%			97.0-103.0

 

Height at 100% composition of B (H₁₀₀) = __________

Observation table for Channel C and D

Eluent Composition	Hx	Calculated    Dx (%)	Acceptance Criteria
20%			19.4-20.6
40%			38.8-41.2
60%			58.2-61.8
80%			77.6-82.4
100%			97.0-103.0

 

Height at 100% composition of D (H₁₀₀) = __________

 

Calculation:

The actually obtained mobile phase composition can be calculated from the Hx values by the following formula.

 

B_X or D_X (%) = H_X (mm)/H₁₀₀ (mm)————— x (100)

= _____________________

Where:

X         =          for the programmed step height (20, 40, 60 and 80).

H_X          =          Height of the respective plateaus in mm of B or D

D_X       =          the actual obtained mobile phase composition.

H₁₀₀       =          Height at 100% composition of B or D.

 

2. Calibration of Auto Injector:-

 

• Vial Test

 

Sr No	Vial Position/ Location	Observation
1		Septum pierced/Not pierced
2		Septum pierced/Not pierced
3		Septum pierced/Not pierced
4		Septum pierced/Not pierced
5		Septum pierced/Not pierced

  

• Volume accuracy of injection:-

 

Instrument/Equipments/Standards used:

Balance No
Balance calibration due date
Injection Volume	20µl
Number of Injections	10
Density of Water at 200C (d)

 

  

Observation Table:

Initial weight of Vial + Water(W1) in gram
Weight of Vial + Water after 10 Injection(W2) in gram
W = (W1-W2) (in gram)
mL of water injected = W/d  in mL
mL of water injected in 10 injection = W/d *1000 µl=A
µl per injection=A/10

  

Acceptance Criteria:

The amount of water drawn by the auto injector per injection should be in between 19.0 µl and 21.0µl (i.e., ± 1.0 µl of the set value)

 

• Injector Linearity:-

Instrument/Equipments/Standards used:

Balance No
Balance calibration due date
Standard used
Standard weight
Standard Lot number
HPLC Column  details

 

 Preparation of Standard Solution (20 ppm Caffeine):

1. Weighed ______ mg of Caffeine in 100 mL volumetric flask, dissolved and diluted up to the mark with Mobile Phase.
2. Further dilute 10mL of this solution to 100mL with Mobile Phase.

                                                        

Observation Table

 

Sr. No	Injection Volume	Area
1	5 µl
2	10 µl
3	20 µl
4	50 µl
5	100 µl
Correlation coefficient(r2) (Acceptance criteria  r2 ≥ 0.999)

  

• Injector Precision-

Instrument/Equipments/Standards used:

Balance No
Balance calibration due date
Standard used
Standard weight
Standard Lot number
HPLC Column  details

  

Preparation of Standard Solution (20 ppm Caffeine):

1. Weighed ______ mg of Caffeine in 100 mL volumetric flask, dissolved and diluted up to the mark with Mobile Phase.
2. Further dilute 10mL of this solution to 100mL with Mobile Phase.

                       

                                    Observation Table

 

Sr. No.	Area
1
2
3
4
5
6
Average Area
RSD (%)

            Acceptance Criteria: RSD NMT 2.0 %             

 

 

• Carryover Test :-

Instrument/Equipments/Standards used:

Balance No
Balance calibration due date
Standard used
Standard weight
Standard Lot number
HPLC Column  details

 

Preparation of Standard Solution (500 ppm Caffeine):

1. Weighed ______ mg of Caffeine in 10 mL volumetric flask, dissolved and diluted up to the mark with Mobile Phase.

                       

                             Observation Table

 

Name	Area	% Carryover in Blank
Blank-1		NA
Sample		NA
Blank-2
Sample		NA
Blank-3
Sample		NA
Blank

                         Acceptance Criteria: NMT 0.02 %  

 

(Area response of blank-2) – (Area response of blank-1)

Carryover % =   ——————————————————— X 100

Average peak area of caffeine obtained from                                                    standard solution

Carryover % =   ———————————————————

 

3. Calibration of Detector:-

 

• Wave Length Accuracy:-

Instrument/Equipments/Standards used:

 

Preparation of Standard Solution (20 ppm Caffeine):

1. Weighed ______ mg of Caffeine in 100 mL volumetric flask, dissolved and diluted up to the mark with Mobile Phase.
2. Further diluted 10mL of this solution to 100mL with Mobile Phase.

 

Observation Table for wavelength 205 nm

Wavelength nm	Response		Wavelength Maxima/ Maximum response at nm	Acceptance Criteria
	Area	Height
201				205nm ± 2 nm
202
203
204
205
206
207
208
209

 

Observation Table for wavelength 245 nm

Wavelength nm	Response		Wavelength minima/ Minimum response at nm	Acceptance Criteria
	Area	Height
241				245nm ± 2 nm
242
243
244
245
246
247
248
249

                                                  

                                 Observation Table for wavelength 273 nm

Wavelength nm	Response		Wavelength Maxima/ Maximum response at nm	Acceptance Criteria
	Area	Height
269				273nm ± 2 nm
270
271
272
273
274
275
276
277

 

 

• Detector Linearity:-

Instrument/Equipments/Standards used:

 

Preparation of Standard Solution (0.5mg/mL Caffeine):

Weighed ______ mg of Caffeine in 50 mL volumetric flask, dissolved and diluted up to the mark with Mobile Phase.

• 0.25mg/mL standard Solution:

Diluted 1mL of stock solution to 20mL with mobile phase

• 0.5mg/mL standard Solution:

Diluted 2mL of stock solution to 20mL with mobile phase

• 100mg/mL standard Solution:

Diluted 4mL of stock solution to 20mL with mobile phase

• 125mg/mL standard Solution:

Diluted 5mL of stock solution to 20mL with mobile phase

• 250mg/mL standard Solution:

Diluted 10mL of stock solution to 20mL with mobile phase

• 500mg/mL standard Solution:

Use stock solution

 Observation Table:

Theoretical Conc. (mg/ ml)	Actual Conc. (mg/ ml)	Area of injection 1	Area of injection 2	Mean Area	Correlation Coefficient
0.025
0.050
0.100
0.125
0.250
0.500

 

     Acceptance Criteria:  Correlation coefficient ≥ 0.999

 

• Lamp Energy:-

 

Acceptance Criteria and results for DAD:

 

Description	Acceptance criteria	Results	Status Pass/Fail
Lowest intensity in range 190 nm to 220 nm	Not less than 2000 cts.
Lowest intensity in range 221 nm to 350 nm	Not less than 5000 cts
Lowest intensity in range 351 nm to 500 nm	Not less than 2000 cts
Lowest intensity in range 501 nm to 950 nm	Not less than 4000 cts
Highest intensity in range 190 nm to 350 nm	Not more than 450000 cts
Highest intensity in range 700 nm to 950 nm	Not more than 300000 cts
Highest intensity for D2 alpha line	Not more than 1200000 cts

 

 Acceptance Criteria and results for VWD:

 

Description	Acceptance criteria	Results	Status Pass/Fail
Highest intensity	More than 10000 cts
Lowest  intensity	More than 200 cts
Average  intensity	More than 5000 cts

 

4. Calibration of column Compartment:-

 

Observation Table for Right side Compartment

Set Temperature (in °C)	Displayed Temperature on instrument ( in° C)	Observed Temperature on thermometer ( in° C)	Difference between set temp and observed temp on thermometer ( in° C)
25.0
40.0
60.0
80.0

 

Observation Table for Left side Compartment

 

Set Temperature (in °C)	Displayed Temperature on instrument ( in° C)	Observed Temperature on thermometer ( in° C)	Difference between set temp and observed temp on thermometer ( in° C)
25.0
40.0
60.0
80.0

 

Acceptance Criteria:

All the observed thermometer temperature values are within ± 2°C of set value.

 

5. Calibration of Autosampler Temperature:-

 

Observation Table

 

Set Temperature (in °C)	Displayed Temperature on instrument ( in° C)	Observed Temperature on thermometer ( in° C)	Difference between set temp and observed temp on thermometer ( in° C)
25.0
20.0
10.0
4.0

 

Acceptance Criteria:

All the observed thermometer temperature values are within ± 2°C of set value.

 

Remark: System complies/does not comply with respect to above test.

 

 

Performed By:                                                         Checked By:

 

                Date:                                                                     Date:

 

 

 

 

 

Calibration of Analytical Balance-Monthly

 

 Test for Uncertainty:

Standard Weight as per current certified weight (0.5g): ________g         Limit = NMT 0.001

No. of Observations	Weight in gm observed	( xi–  x )2
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.

Σ (x_i – x  ) ² =________

SD    =              √ Σ  ⁿ ( x _i – x)²       =__________

                              ⁱ⁼¹           n -1

SD =   3 X SD of 10 Replicate weighing = ________________

Standard weight= ______________________

SD= Standard Deviation

2. Checks For Off Centre and Shift Error:

    Standard Weight as per current certified weight (0.05g): _________g

    Limit = Error NMT 0.5 mg      

Position of Weight

        ♦1                                      ♦   2

                            ♦ 5

        ♦ 3                                        ♦  4

  

Sr. No.	Weight in gm observed	Error  (NMT 0.5 mg)
1
2
3
4
5

 

3. Sensitivity Check: Acceptance Criteria: + 0.1%

 

Sr. No.	Nominal Weights in  gm	Standard Weights in gm (Certified)	Acceptance Criteria w.r.t std. weights in gm	Observation in gm
1.	100.0000 g
2.	50.0000 g
3.	20.0000 g
4.	10.0000 g
5.	5.0000 g
6.	2.0000 g
7.	1.0000 g
8.	0.5000 g
9.	0.2000 g
10.	0.1000 g
11.	0.0500 g
12.	0.0200 g

 

The results Complies / Does not complies as per acceptance criteria.

 

Performed By: ________________         Checked By:__________

Date: ________________                                      Date: _________

 

 

“End of Document”

Calibration of Analytical Balance-Daily

  

Standard Weights	Certified value	Sensitivity Range
0.0200 g
0.1000 g
1.0000 g
50.0000g
100.0000g

 Performance Check:-                                                                             

Acceptance Criteria: +0.1%

Standard weight box ID no.  _____________

 

Date	Zero Error	Standard Weights					Remark	Done by	Checked by
		0.0200 g	0.1000 g	1.0000 g	50.0000g	100.0000g

End of Document