all sops combined pdf.pdf

PRACTICE SCHOOL GROUP -V Page 1
YB Chavan college of pharmacy
Aurangabad
TITLE:-OPERATION AND OF U.V. VISIBLE
SPECTROPHOTOMETER
Objective To study procedure for operating and calibrating u.v. Visible
spectrophotometer.
Scope – The procedure is applicable to operate and calibrate u.v. Visible
spectrophotometer.
Instrument:-
Page no. 1 of 1
SOP no. 1
Revision
no.
0
Created by Shaikh Asif Jamil
Created date 22/10/2021
Location Machine room
Review Date 22-4-22
Revision No.
Revisor
Responsibility Barrawaz Ateka Yahiya
Accountability
HOD OF Q.A.Dept
Name U.V visible spectrophotometer
Maker SHIMDZU
Model U.V.-1780

Requirement – 2cuvetts, tissue paper, DOUBLE BEAM
SPECTROPHOTOMETER
Operation procedure-
1)Ensure the power plug of instrument and computer is not on.
2) Switch on the power supply pressing main switch of switchboard.
3) Remove silica bag from panel door.
4) On the system and instruments power buttons beep sound shows indication of
on.Give some time to initialize the machines.
5) Make sure cuvettes are crackless and clean on both sides.
(Lambda max of cuvettes should be zero).
6) Fill both the cuvettes with same blank sample and place in the cuvettes holder
facing the blurred side facing yourself (clean if needed)
7) Open the UV probe version 2.70 software in the adjacent computer system.
8) Click “connect” button on bottom left side of screen the initializing will start in
series showing green dots serially as LSI initialization , WI lamp energy check,
wavelength origin 1 check, wavelength origin 2 check, standby
9) Now, put dilution sample in both cuvetts and close the instrument panel door.
10) Click on connect button on system and click on auto zero to tare the dilution
sample to 0 absorbance.
11) Now change the closely placed cuvett with different samples to analyse keeping
the upper side cuvett filled with dilution solvent sample .
12) The spectrophotometer is ready for further use.
Caution-
1) Always remove silica bag before use of spectrophotometer and put bafore leaving
machine room.
2) Always fill cuvettes at least 90%.
3) Close panel lead before running sample.
4) Before putting switch buttons off make sure the system and instrument is off already.
5) Wait after startimg the system to warm the sources of light.
Page no. 2of 2
SOP no. 1
Revision
no.
0
CREATED BY :- APPROVED
BY:-
REVIEWED
BY:-
AUTHORISED
BY :-
Shaikh Asif jamil Ms. Barrawaz
Aateka Y
HOD Dr. JP
Sanghshetti
Dr. Abubakar
Salam Bawazir
DATE &
SIGNATURE
28-10-2021
BAY JNS ASB

Aim: To study the effect of concentration on absorbance.
Reference:
 Chatwal GR, Anand S (2002) Instrumental Methods of Chemical Analysis. (5th
ed),
Himalaya Publishing House, New Delhi.
 Davidson AG (2002) Ultraviolet-visible absorption spectrophotometer. In Beckett
AH, Stenlake JB, (4thedn), Practical Pharmaceutical chemistry. CBS Publishers and
distributors, New Delhi, 275-278.
 ICH (1996) Q2B Validation of Analytical Procedures-Methodology. Consensus
Guidelines, ICH Harmonized Tripartite Guidelines
Requirements:
Glassware: Beakers, volumetric flasks, pipette, cuvettes
Chemicals: Paracetamol, Methanol, Distilled water
Instrument: SHIMADZU 1780
Software: UVProbe 2.70
Theory:
UV spectroscopy is an absorption spectroscopy in which the molecules absorb
wavelengths in UV region (400nm- 200nm) resulting in excitation of electrons. The
instrument used is UV spectrophotometer which consists of components like radiation
source, monochromator, detector, a recording system and sample cell. The instrument is
connected to a software. The software measures the wavelength of samples of known
concentration at which maximum absorbance is observed. At this maximum wavelength
the absorbance of sample measured.
Beer’s law: It states that the intensity of a beam of parallel monochromatic radiation
decreases exponentially with the number of absorbing molecules. In other words,
absorbencies proportional to the concentration.
Lambert’s law: It states that the intensity of a beam of parallel monochromatic
radiatiodecreases exponentially as it passes through a medium of homogeneous thickness.
A combination of these two laws yields the Beer-Lambert law.
Beer-Lambert law: When beam of light is passed through a transparent cell containing a
solution of an absorbing substance, reduction of the intensity of light may occur.
Mathematically, Beer- Lambert law is expressed as
A = a. b. c
Where, A = Absorbance or optical density
a = Absorptivity or extinction coefficient
b = Path length of radiation through sample
c = Concentration of solute in solution.
Both b and a are constant so a is directly proportional to the concentration
Procedure:
Procedure I: Preparation of Solutions
Preparation of Stock solution (100μg/ml):
Weigh accurately about 10 mg paracetamol and dissolve it in 15 ml methanol in 100 ml
volumetric flask, make the volume upto 100 ml with distilled water.

PREPARATION OF BLANK: Prepare 15: 85 %v/v solution of methanol and distilled
water, as a blank and as a dilution solvent for standard dilution series.
PREPARATION OF SAMPLE SOLUTIONS OF DIFFERENT CONCENTRATIONS:
Pipette out 1, 2 to 5 ml of stock solution in 10 ml of volumetric flasks & make the volume
upto the mark with dilution solution to get 10, 20, to 50 µg/ml respectively.
Procedure II:
Turn ON the UV-Vis Spectrophotometer and connected system.
Run the UV Probe 2.70 software.
Wait till the instrument to be initialised properly.
Open Spectrum module in software.
Wash the cuvettes with distilled water and rinse with the blank solution.
Fill the cuvettes ≈ 80% with blank solution & clean it with lint free tissue paper.
Place cuvettes in the sample compartment, and click on the Auto Zero button.
Select the wavelength (nm) range to run the spectra.
Wash and rinse the sample with the sample solution, scan the solution to get absorbance
maxima (λmax).
Calculate the absorbance maxima (λmax) for each dilution, andinterpreter the results for
effect of concentration.
Observation: Table 2.1 Absorbance of paracetamol in methanol
Concentration (µg/ml) λmax (nm) Absorbance
10 244.40 0.271
20 244.40 1.862
30 244.40 2.677
40 244.40 3.454
50 244.40 3.844
Fig. 2.1: Spectrum on 50 µg/ml Fig. 2.2: Spectrum on 40 µg/ml
nm.
200.00 250.00 300.00 350.00 400.00
Abs.
4.383
4.000
3.000
2.000
1.000
0.000
-0.254
nm.
200.00 250.00 300.00 350.00 400.00
Abs.
4.387
4.000
3.000
2.000
1.000
0.000
-0.253

Fig. 2.3: Spectrum on 30 µg/ml Fig. 2.4: Spectrum on 20 µg/ml
.Fig. 2.5: Spectrum on 15 µg/ml Fig. 2.6: Overlay spectra
All the recorded spectra reveals that as we decrease the concentration of absorbing
substance in the solution the relative absorbance also decreases.
Result:
Overlay spectra assures the Beer’s Law which states, the concentration is proportional to
absorbance of the solution.
Conclusion:
The prepared dilution series obeys the Beer’s Law.
nm.
200.00 250.00 300.00 350.00 400.00
Abs.
4.387
4.000
3.000
2.000
1.000
0.000
-0.253
nm.
200.00 250.00 300.00 350.00 400.00
Abs.
2.569
2.000
1.000
-0.092
nm.
200.00 250.00 300.00 350.00 400.00
Abs.
3.387
3.000
2.000
1.000
0.000
-0.167
nm.
200.00 250.00 300.00 350.00 400.00
Abs.
4.387
4.000
3.000
2.000
1.000
0.000
-0.254

Aim: To determine concentration of unknown solution using standard solution.
Reference:
Chatwal GR, Anand S (2002) Instrumental Methods of Chemical Analysis. (5thedn),
Davidson AG (2002) Ultraviolet-visible absorption spectrophotometry. In Beckett AH,
Stenlake JB, (4thedn), Practical Pharmaceutical chemistry CBS Publishers and
distributors, New Delhi, 275-278.
Requirements:
GLASSWARE: Beakers, volumetric flasks, pipette, cuvettes
CHEMICALS: Acetyl salicylic acid (Aspirin), Ethanol, Distilled water
INSTRUMENT: SHIMADZU 1780
SOFTWARE: UVProbe 2.70
Theory: Refer to experiment no.1 page no.1-3
Procedure:
Procedure I: Preparation of Solutions
PREPARATION OF STOCK SOLUTION (100ΜG/ML):
Weigh accurately about 10 mg aspirin and dissolve it in 10 ml ethanol in 100 ml
volumetric flask, make the volume upto 100 ml with distilled water.This gives a solution
of concentration 1mg/ml. Dilute 2ml from this solutionin20ml ethanol.Thisgives
asolutionofconcentration 100µg/ml.
Procedure II:
Turn ON the UV-Vis Spectrophotometer and connected system.
Run the UV Probe 2.70 software.
Wait till the instrument to be initialised properly.
Open Spectrum module in software.
Wash the cuvettes with distilled water and rinse with the blank solution.
Fill the cuvettes ≈ 80% with blank solution & clean it with lint free tissue paper.
Place cuvettes in the sample compartment, and click on the Auto Zero button.
Select the wavelength (nm) range to run the spectra.
Wash and rinse the sample with the sample solution, scan the solution to get absorbance
maxima (λmax).
Open Photometric module to determine absorbance of the solutions at absorbance
maxima (λmax) for each dilution.
Calculate the concentration of unknown sample for each dilution

Observation: Table: 3.1Absorbance of Aspirin in ethanol and water
Concentration (µg/ml) Absorbance
20 0.142
30 0.248
40 0.434
50 0.537
Unknown Sample 0.068
Calculations:
Accordingto Beer-Lambertlaw; A = ɛ c t
Where, ɛ and t are constant
Let,
A1 =the absorbance of standar dsolution of known concentration.
A2 = the absorbance of solution of unknown concentration.
C1= the concentration of standard solution
C2=th econcentration of sample
By comparing the above equation for known and unknown samples, the
equation can be written as follows,
=
i.e. C2 = × C1
Consider, C1 =20µg/mlandA1=0.142
C2 = × 20 = 9.577µg/ml
Consider, C1 = 30µg/mlandA1=0.248
C2 = × 30 = 8.225µg/ml
Consider, C1= 40µg/mlandA1=0.434
C2 = × 40 = 6.267µg/ml
Consider, C1= 50µg/mlandA1=0.537
C2 = × 50 = 6.331µg/ml
Mean concentration of unknown sample = = 7.60µg/ml
Result:

The concentration of unknown sample was found to be 7.60 µg/ml.
Conclusion:
Determination of concentration of unknown solution using standard solution by UV-Vis
Spectrophotometer was done.

Aim: Determination of concentration of unknown sample by using calibration curve
method.
Reference:
Gurdeep R. Chatwal, Sham K. Anand Instrumental Methods of Chemical Analysis,
Himalaya Publication Fifth Edition (Reprints 2015), page no.2.1082& 2109
H. Willard - Hobert, L Merritt Jr Lynne John , A, Sttle Jr. Frank, “ Instrumentation
Method Of Analysis” CBS Publishers and Distributors ,New Delhi.
Requirements:
SAMPLE: Paracetamol
DILUTION SYSTEM: Methanol and Water (15:85 %v/v) used as diluents.
INSTRUMENTATION: SCHIMADZU UV-1780 UV-Visible double beam
spectrophotometer with matched quartz cells.
Theory: Refer to experiment no. 01 page 1-3
Procedure: I
PREPARATION OF STOCK SOLUTION (100ΜG/ML):
Weigh accurately about 10 mg aspirin and dissolve it in 10 ml ethanol in 100 ml
volumetric flask, make the volume up to 100 ml with distilled water. This gives a solution
of concentration 1mg/ml. Dilute 2ml from this solutionin20ml ethanol. This gives a
solution of concentration 100µg/ml.
Procedure: II Refer to experiment 02.
Observation:
Absorbance maxima of Paracetamol in methanol- water :
Figure 4.1 : λmax 2.44.40nm
nm.
200.00 250.00 300.00 350.00 400.00
Abs.
3.387
3.000
2.000
1.000
0.000
-0.167

Observation Table:4.1 ABSORBANCE OF PARACETAMOL IN METHANOL
AND WATER
Sr. no. Concentration ( µg/ml ) Absorbance
1 2 0.070
2 4 0.141
3 6 0.161
4 8 0.187
5 10 0.271
6 12 1.158
7 15 1.404
8 20 1.862
9 Unknown 1.695
Calculation:-fig 4.2Determination of unknown concentration by calibration curve
method
Y = 0.0883 x - 0.1932
0.088 x = 1.695 + 0.193
0.088 x = 1.888
x = 1.888/0.088
x = 21.45 µg/ml
Result: Concentration of Unknown sample was found to be 21.45 µg/ml and its
absorbance was 1.695.
y = 0.111x - 0.4112
R² = 0.8749
-0.5
0
0.5
1
1.5
2
0 10 20 30
absorbance
absorbance
Linear
(absorbance)

Aim: To determine of Absorption maxima of the given sample
Reference:
Chatwal GR, Anand S (2002) Instrumental Methods of Chemical Analysis. (5th
edn),
Davidson AG (2002) Ultraviolet-visible absorption spectrophotometry. In Beckett AH,
Stenlake JB, (4thedn), Practical Pharmaceutical chemistry. CBS Publishers and
distributors, New Delhi,275-278.
Requirements:
GLASSWARE: Beakers, volumetric flasks, pipette, cuvettes
CHEMICALS: Metformin, Methanol, Distilled water
INSTRUMENT: SHIMADZU1780
SOFTWARE: UV Probe2.70
Theory:Absorption maxima (λmax): The extent to which a sample absorbs light
depends on the wavelength of light. The wavelength at which a substance shows
maximum absorbance is called Absorption maxima or λmax
Procedure:
Fill both the cuvettes with the blank (solvent).
Clean both the cuvettes with tissue paper and place them in the sample cell such that the
blur side faces the user.
In the SPECTRUM mode click on AUTO ZERO. This records the absorbance of
solvent as 0.
Replace the front cuvette with the sample under analysis.
Select the wavelength range for UV that is 400nm-200nm and click on START
In MAIN MENU select PICK PEAK to select the desired peak wavelength
Spectra: Fig. 5.1 Absorption Maxima of Metformin
Conclusion: Absorption Maxima (λmax) of Metforminwas found to be 232.6 nm with

absorbance 1.924
OBJECTIVE: - To describe the procedure for handling of UV- Visible
Spectrophotometer
SCOPE:- The procedure is applicable for single beam spectrophotometer.
INSTRUMENT:-
Instrument Single beam spectrophotometer
Maker Chemito
MODEL UV2100
Dr. RafiqZakaria
Y.B. Chavan College of Pharmacy, Aurangabad
TITLE:- STANDARD OPERATING PROCEDURE FOR HANDLING OF
SINGLE BEAM UV- VISIBLE SPECTROPHOTOMETER.
Page no. 1 of 3
SOP no. 2
Revision
no.
0
DEPARTMENT:- Quality Assurance&
Quality
control
LOCATION:- Chemistry lab
DATE:- 25-11-2021
CREATED BY:-roll 99 REVISION BY :-
REVIEW DATE:-25-02-2022
LOCATION:- chemistry lab AUTHARISED BY:-

PROCEDURE:-
 Connect the instrument to power supply .
 Switch on the main power supply
 The yellowish light flickers will glow one by one .
Check the initialization as-
o ROM test
o RAM test
o Monochromatar
o Filter check
o Turret check
o Optimising W. lamp
o Optimising D2 lamp
o Lamp Stability
 after complete initialization is done
 open the main menu there will be three options,
 Spectrum mode
 Photometric mode
 Select Mode
 Now open the sample holder lid. There will be 5 cuvette holders as shown below.
 To the blank using distilled water.
 Then fill analyte in the cuvette.
 Select the spectrum mode or photometric mode.
 To have to find absorbance then select the photometric mode.
 Then press ESCAPE button.
 Select the cuvette no. In which the sample was placed.
Page no. 2 of 3
SOP no. 2
Revision
no.
0

Causion:-
1)always remove silica bag before use of spectrophotometer and put before leaving
machine room
2) always fill cuvetts at least 90%.
3) close panel lead before running sample.
4) before putting switch buttons off make sure the system and instrument is off already.
5) wait after startimg the system to warm the source bulbs.
Page no. 3 of 3
SOP no. 2
Revision
no.
0
BY:-
REVIEWED
BY:-
AUTHORISED
BY :-
Aateka Y
HOD Dr. JP
Sanghshetti
Dr. Abubakar
Salam Bawazir
DATE &
SIGNATURE
25-11-2021
BAY JNS ASB

Aim :- Preparation of calibration curve of Metformin.
Reference:
1. Gurdeep R. Chatwal, Sham K. Anand Instrumental Methods of Chemical Analysis,
Himalaya Publication Fifth Edition(Reprints 2015), page no.2.108 & 2.109
2. United State pharmacopoeia, National formulary-35,2017, 40,4415,4418
http://www.drugs.com/pro/metformin-tablet.html
Requirements:-
Sample: Metformin
Dilution system : distilled water used as diluent
Instrumentation: “ CHEMITO ” UV2100 UV - Visible Spectrometer ( single beam)
Theory:- Refer previous experiment pege no,13-14
Procedure:- Sampling and Dilution procedure is Same as previous experiments.
Operation Procedure:-
Turn on the UV- Visible Spectrophotometer.
The yellowish light flickers and the instrument starts INISITIALIZATION as under;
o ROM test
o RAM test
o Monochromator
o Filter check
o Turret check
o Optimising w.lamp
o Optimising D2 lamp
o Lamp stability
Open the sample holder lid.
Perform the blank spectra using distilled water in photometric mode.
Now place the metformin sample in sample holder place the transparent side of
cuvette to screw side to pass the light source.
Select the photometric mode then select the required wavelength i.e. 276nm for
metformin
Press escape button.
Press the cuvette no in which our sample was placed.
Note down the absorbance reading.
Observation table:- Table 7.1 Absorbance of metformin in water
Sr.
No
Concentration µg/ml Absorbance
2. 24 2.046
3. 20 1.886
4. 16 1.678
5. 12 1.215
6. 8 0.907
7. 4 0.590

Wavelength = 276 nm
Figure :7.1 Calibration curve of metformin
Result:- Calibration Curve of Metformin was prepared.
0
0.5
1
1.5
2
2.5
0 10 20 30
Absorbance
concentration
Absorbance
Linear (Absorbance)

Aim:-Comparison of calibration curve prepared using single & beam UV- Visible
Spectrophotometer.
Reference:
1. Gurdeep R.Chatwal ,Sham K. Anand Instrumental Methods of Chemical Analysis,
Himalaya Publication Fifth Edition(Reprints 2015),page no.2.108&2.109
2. United State pharmacopopiea, National formulary-
35,2017,40,4415,4418http://www.drugs.com/pro/metfrmin-tablet.html.
Requirements:
Sample: Metformin
Dilution system: distilled water used as diluent
Instrumentation:
1) SCHIMADZU UV-1780 UV-Visible double beam spectrophotometer
2) CHEMITO UV2100 UV - Visible Spectrometer (single beam)
Theory:Refer experiment no:3 page 10-12
Procedure:Refer experiment no:3 page 10-12
Observation: ⅄ max276nm
Table 8.1: Comparison of absorption by Chemito and Shimadzu Spectrophotometer
conc. Abs by CHEMITO Abs by SHIMADZU
4 0.59 0.425
8 0.907 0.741
12 1.215 1.152
16 1.678 1.566
20 1.886 1.924
Figure8.1: Comparison of calibration curve of Metformin by Chemito and
Shimadzu Spectrophotometer
Result: Comparison of calibration curve prepared using single (CHEMITO)&double
(SHIMADZU 1780) beam UV- Visible Spectrophotometer was done.
y = 0.0841x + 0.2463
R² = 0.9903
y = 0.0956x + 0.0147
R² = 0.9981
0
0.5
1
1.5
2
2.5
0 5 10 15 20 25
Abs by Chemito Abs by shimadzu
Linear (Abs by Chemito) Linear (Abs by shimadzu)

Dr. RafiqZakaria
Y.B. Chavan College of Pharmacy, Aurangabad
TITLE: - STANDARD OPERATING PROCEDURE FOR HANDLING
OF FTIR
OBJECTIVE:- To describe the procedure for handling of FTIR.
SCOPE:- To design the SOP for operation of FTIR spectrophotometer.
INSTRUMENT:-
Page no. 1 of 3
SOP no. 3
Revision
no.
0
Review Date 22-4-22
Accountability HOD OF Q.A.Dept
Page no. 2 of 3
SOP no. 3

PROCEDURE:
 Clean the instrument
 Ensure that the power to the instrument is switched OFF before cleaning Clean
the instrument with a clean dry cloth daily.
 Remove silica bag from it.
 Ensure that the instrument is properly connected to the power supply.
 Switch ON the main switch and power switch of the instrument.
 Switch the computer ON by the switches situated on the right of the monitor and
PC respectively.
 Switch on the instrument, keep it for 5 to 10 mins to get warm.
 Click on software named ‘spectra manager’ & select collect mode
 In collect mode select background & put file name or as per your requirement (in
case of DRA assembly put KBr powder).
 Now click on ‘ok collect’ so that it will scan background spectra.
 After completion of background spectra, put sample pellet (in case of DRA put the
mixture of KBr & sample).
 Again click on collect & select normal, transmittance mode, select scanning range
(4000-5000 cm-1 or as per your requirement). Select number of scan.
 After completion of scanning you will get a spectrum of the actual sample.
 For peak marking click on view & select ‘peaksfind’
Revision
no.
0
Instrument FTIR
Maker JASCO
MODEL 4100

 For comparison with your previous standard just right click on main
screen & select overlaid or stacked mode as per your requirement
 Now open your previous standard spectrum by click on file – open trace.
 You can also add the data on the screen by right click on the main screen & select
add text.
 Save the data.
 For Solids (hip method ):-
 Take pre heated KBR as solvent with sample
 Take minute amount of sample with spatula mix with KBR in small mortar pestle
.
 Mix sample + KBR.
 Put it in the cuvette with the pestle.
 Put the cuvette in the cuvette holder.
 See the red lazer dot on the sample for reference to the source light.
o Save the peak data in word format
 Caution-
 1)always remove silica bag before use of spectrophotometer and put before
leaving machine room
 2) Always fill cuvettes at least 90%.
 3) close panel lead before running sample.
 4)before putting switch buttons off make sure the system and instrument is off
already.
 5) wait after starting the system to warm the source bulbs.
Page no. 3 of 3
SOP no. 3
Revision
no.
0
BY:-
REVIEWED
BY:-
AUTHORISED
BY :-
Aateka Y
HOD Dr. JP
Sanghshetti
Dr. Abubakar
Salam Bawazir
DATE &
SIGNATURE
22-10-2021
BAY JNS ASB

Y.B. Chavan college of pharmacy,
Aurangabad
TITLE:- calibration of FTIR spectrophotometer
OBJECTIVE:- To describe procedure for calibration of FTIR
spectrophotometer.
SCOPE:- Applicable for calibration of FTIR spectrophotometer.
INSTRUMENT:-
Instrument FTIR
Maker Jasco
MODEL 4100
PROCEDURE:-
1) WITH POLYSTYRENE SHEET
 Open the FTIR panel cover unscrew the cuvette holder.
 Replace it with polystyrene holding panel with a screw.
 Put the polystyrene sheet in the holder
 Click on ‘peak find’, it will give downwards graph peaks of wavenumber v/s %T.
 Campare the peaks with the standard IP polystyrene peaks
 If the noise levels are very high minimize it and again click on
apply to again measure graph for compare with standard peaks
as shown below.
Page no. 1 of 3
SOP no. 4
Revision
no.
0
Review Date 22-4-22
Revision No.
Revisor
Accountability
HOD OF Q.A.Dept
Page no. 2 of 3
SOP no. 4
Revision
no.
0

Fig.10.1 Transmittance wavelength curve for polystyrene
 if the peak matches it passes the calibration.
 Wave number and %T values considered are as
Table 10.1 Wave length and acceptable tolerance of IR
Wave number (cm-
1
)
Acceptable tolerance Percent
transmittance value
(cm-1
)
Remarks
3060.0 ± 1.0 3059.51 Satisfactory
2849.5 ± 1.0 2849.31 Satisfactory
1601.2 ± 1.0 1600.63 Satisfactory
1583.0 ± 1.0 1583.24 Satisfactory
1154.5 ± 1.0 1154.19 Satisfactory
1028.3 ± 1.0 1027.87 Satisfactory
Match wavenumber with %transmittance and calibrate with comparing.
PRECAUSIONS:-
1. Use only spectroscopic grade KBR ie dried thoroughly
2. Clean and dry mortar and pestle immediately after use .
3. Polystyrene film should be kept in protecting cover.
4. always remove silica bag before use of spectrophotometer and put before leaving
machine room
5. close panel lead before running sample.
6. before putting switch buttons off make sure the system and instrument is off
already.
7. wait after starting the system to warm the source bulbs.

Page no. 3 of 3
SOP no. 4
Revision
no.
0
BY:-
REVIEWED
BY:-
AUTHORISED
BY :-
Aateka Y
HOD Dr. JP
Sanghshetti
Dr. Abubakar
Salam Bawazir
DATE &
SIGNATURE
28-10-2021
BAY JNS ASB

Dr. Rafiq Zakaria Campus
YB CHAVAN COLLEGE OF PHARMACY AURANGABAD
CALIBRATION OFCHEMITO 2100 AND SHIMADZU 1780
UV VISIBLE SPECTROPHOTOMETER
DEPARTMENT: Qualityassurance& Analysis AREA: Instrument room
SOP: PS-04 SUPERCEDES: --
Eff. Date:04/12/2021 Review date:04/06/2022
01. Objective:To lay down a procedure for uv spectrophotometer
02. Responsibility:All personnel of Quality Assurance and Analysis Department
03. Accountability:HOD, Quality Assurance and Analysis Department
04. Main features:
4.1 Name of the instrument: UV VISIBLE SPECTROPHOTOMETER
4.2 Instrument serial no. MAET/02/03
4.3 Make and Model: CHEMITO 2100 and SHIMADZU 1780
4.4 Manufacturer: CHEMITO and SHIMADZU
05. Calibration
Calibration of UV- VIS spectrophotometer involves:
5.1 Control of Absorbance
5.2 Limit of Stray Light
5.3 Resolution Power
5.4 Cells
5.5 Solvents
5.1 Control of absorbance:
5.2.1 Preparation of Solution A and Solution B: For solution A accurately weigh 57-63 mg
of potassium dichromate and dissolve it in 100 ml of 0.005 m Sulphuric acid. From this
solution take 1ml and dilute it in 10 ml of 0.005 M Sulphuric acid for solution B.
5.2.2 Fill one cuvette with 0.005 M sulfuric acid
5.2.3 In MAIN MENU click on Key 2 (PHOTOMETRIC) to record the absorbance of
blank.
5.2.4Place the solution in sample cell with potassium dichromate solution.
5.2.5 Go to PHOTOMETRIC MODE and determine the absorbance at the first four
wavelengths mentioned in Table 1. For solution A
5.2.6 Now determine the absorbance for solution B at 430 nm
5.2.7 Note the absorption maxima of potassium dichromate solution at different wavelengths
and calculate the absorbance.
5.2.8 The permitted tolerance is given in the table below.
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SOP no. 5
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Table 11.1 Standard Absorptivity and maximum tolerance
Sr. no Wavelength Maximum
Tolerance
1 235 nm 122.9 to 126.2
2 257 nm 142.8 to 146.2
3 313 nm 47.0 to 50.3
4 350 nm 105.6 to 109.0
5 430 nm 15.7 to 16.1
OBSERVATION: table 11.2 Absorbance Value by CHEMITO and SHIMADZU
Sr.
no Wavelength
Observed
valueAbsorbance
Absorptivity Result
By
Chemito
By
Shimadzu
By
Chemito
By
Shimadzu
By
Chemito
By
Shimadzu
1 235 nm 0.706 0.7410 117.6 123.5 PASS Pass
2 257 nm 0.646 0.8620 107.6 143.5 FAIL Pass
3 313 nm 0.233 0.2890 38.83 48.6 FAIL Pass
4 350 nm 0.391 0.6340 65.16 105.3 FAIL Pass
5 430 nm 0.107 0.9531 17.8 15.8 FAIL Pass
5.2 Limit of Stray Light
5.3.1 Prepare a 1.2 % w/v potassium chloride solution.
5.3.2 Fill water in one cuvette and place it in reference cell and in MAIN MENU click on
Key 2 (PHOTOMETRIC MODE) to record the absorbance of blank
5.3.3 Place water in the reference cell and 1.2 % w/v potassium chloride solution in sample
cell 2.
5.3.4 Go to PHOTOMETRIC MODE and determine the absorbance of the solution at 200
nm.
5.3.5 The Absorbance at 200 nm must be greater than 2.
OBSERVATION:
Absorbance at 200 nm: 0.426
Result:Satisfactory
5.3 Resolution power
5.4.1 Prepare a 0.02% v/v solution of toluene in hexane.
5.4.2 Place hexane in the reference cell and in MAIN MENU click on Key 2
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SOP no. 5
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(PHOTOMETRIC) to record the absorbance of
blank5.4.3Place hexane in the reference cell and toluene in
hexane in sample cell 2
5.4.4 Go to SPECTRUM MODE and record the absorbance at
269 nm and 266 nm.
5.4.3 The ratio of absorbance at 269 nm to 266 nm is not less than 1.5
OBSERVATION: table 11.3 Absorbance of n hexane in toulene
By Chemito By Shimadzu 1780
n-hexane
in toluene
269 nm 1.367 1.660
266 nm 1.602 2.296
Ratio 0.85 1.3
Result NotSatisfactory NotSatisfactory
5.4 Cells
5.4.1Determine the absorbance of air in PHOTOMETRIC MODE
by removing the cuvette from the reference cell
5.4.2 Now place an empty cuvette and record its absorbance in PHOTOMETRIC MODE
5.4.3 Determine the ratio of Absorbance of air to Absorbance of cuvette
5.4.4 The tolerance on the path length of the cells used is ± 0.005 cm.
OBSERVATION:
a) Absorbance of Air
b) Absorbance of Cuvette: 0.082
Result: Satisfactory
5.5 Solvents
5.5.1 Take an empty cuvette and place it in the reference cell
5.5.2 In MAIN MENU click on Key 2 (PHOTOMETRIC) and record the absorbance of air
at 254 nm
5.5.3 Fill the other cuvette with ethanol and place it in the sample cell 2 and in MAIN
MENU click on Key 2 (PHOTOMETRIC)to
record the absorbance of ethanol at 254 nm
5.5.4 The absorbance of the reference cell and its contents should not exceed 0.4 and should
preferably be less than 0.2 when
measured with reference to air at the same wavelength
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5.5.5Precaution:The solvent in the reference cell should be of
the same lot as that used to prepare the solution and must be free
from fluorescence at the wavelength of measurement.
OBSERVATION:
table 11.4 Absorbance of ethanol and air
By CHEMITO 2100 By SHIMADZU 1780
Absorbance of air 0.273 0.416
Absorbance of ethanol 3.000 3.73
Ratio 0.079 0.11
Result Satisfactory Satisfactory
Page no. 1 of 4
SOP no. 5
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When the instrument does not comply with the requirement/tolerance specified above, the
instrument must be labelled ‘OUT OF CALIBRATION’ and must be repaired /serviced.
BY:-
REVIEWED
BY:-
AUTHORISED
BY :-
Aateka Y
HOD Dr. JP
Sanghshetti
Dr. Abubakar
Salam Bawazir
DATE &
SIGNATURE
26-11-2021
BAY JNS ASB

DR. RAFIQ ZAKARIA CAMPUS
YB CHAVAN COLLEGE OF PHARMACYAURANGABAD
STANDARD OPERATING PROCEDURE (SOP) AND
CALIBRATION OF ELECTRONIC BALANCE
DEPARTMENT: Qualityassurance& Analysis AREA: Instrument room
SOP: PS-03
Eff. Date:26/11/2021 Review date: 26/05/2022
Rev. No. : - Page no: 1 of 4
01. Objective:To lay down a procedure for Operation ofELECTRONIC BALANCE
02. Responsibility:All personnel of Quality Assurance and Analysis Department
03. Accountability:HOD, Quality Assurance and Analysis Department
04. Main features
4.1 Name of the instrument: ELECTRONIC BALANCE
4.2 Instrument serial no.
4.3 Make and Model:
4.4 Manufacturer: WENSAR
Page no. 1 of 3
SOP no. 6
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Page no. 2 of 3
SOP no. 6
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05. Precautions:
5.1 Ensure that the weighing platform is clean
5.2 Do not disturb the balance while placing or removing the substance to be weighed
5.3
06. Operation:
6.2 Turn on the balance and let it warm up until it displays 0.000
6.3 If the value is found to fluctuate press 0/T to bring the reading to 0.000
6.4 Place a clean butter paper on the weighing platform and place the compound to be
weighed onto the butter paper
6.5 The units in which the weight has to be determined is selected by pressing the
MODE button
6.6 The weight of the compound placed is displayed.
07. Calibration:
7.2 Turn on the balance and let it warm up until it displays 0.000
7.3 If the value is found to fluctuate press 0/T to bring the reading to 0.000
7.4 Load a standard weight on to the weighing balance
7.5 Determine its weight on the weighing balance
7.6 If the value displayed fluctuates from that of the known value, the issue must be
reported to the service engineer and labelled as ‘OUT OF CALIBRATION’

Page no. 3 of 3
SOP no. 6
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0
BY:-
REVIEWED
BY:-
AUTHORISED
BY :-
Aateka Y
HOD Dr. JP
Sanghshetti
Dr. Abubakar
Salam Bawazir
DATE &
SIGNATURE
26-11-2021
BAY JNS ASB

Dr. Rafiq Zakaria Campus
YB CHAVAN COLLEGE OF PHARMACY AURANGABAD
TITLE: STANDARD OPERATING PROCEDURE (SOP) AND CALIBRATION
COSLAB DIGITAL MELTING POINT APPARATUS
Objective: To lay down a procedure for Operation of DIGITAL MELTING POINT
apparatus.
Responsibility: All personnel of Quality Assurance and Analysis Department
Accountability: HOD, Quality Assurance and Analysis
Instrument:
1. Name of the instrument: DIGITAL MELTING POINT APPARATUS
2. Instrument serial no.
3. Make and Model:
4. Manufacturer: COSLAB
Page no. 1 of 3
SOP no. 7
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DEPARTMENT: Quality assurance &
Analysis
AREA: chemistry lab
Subject: SOP for handling and calibration of
digital mrlting point
Eff. Date: 26/11/2021 Review date: 26/05/2022

Precaution:
Clean the capillary and check if sealed completely.
Operation:
1. Switch on the electric supply and the instrument
2. Switch on the start button
3. Switch on the STIRRER and adjust the speed
4. Seal a capillary in flame and fill the powder whose melting point is to be
determined
5. Insert the capillary in the sample holder.
6. Adjust the intensity of the background light such that the inserted capillary as well
the powder in it are clearly visible
7. Adjust the temperature using the HEATER control knob Increase the heat initially
at higher rate; as the melting point approaches, lower the heating rate to observe
the clear melting of solid near its melting temperature range
Calibration:
7.1 For boiling point:
1. Fill the sample holder with a liquid of known boiling point for example water
5. Adjust the intensity of the background light such that the sample tube as well the
liquid in it are clearly visible
6. Adjust the temperature using the HEATER control knob
7. Increase the heat initially at higher rate; as the boiling point approaches, lower the
heating rate to observe the clear vaporization of liquid near its boiling temperature
range
8. If the boiling point is same as that of the known boiling point of standard liquid;
the instrument is calibrated
7.2 For melting point
1. Seal a capillary in flame and fill the powder with known melting point
2. Insert the capillary in the sample holder.
6. Adjust the intensity of the background light such that the inserted capillary as well
the powder in it are clearly visible
7. Adjust the temperature using the HEATER control knob
Page no. 2 of 3
SOP no. 7
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8. Increase the heat initially at higher rate; as the melting point approaches, lower
the heating rate to observe the clear melting of solid near its melting temperature
range
9. If the melting point is same as that of the known melting point of
standard liquid; the instrument is calibrated.
Page no. 3 of 3
SOP no. 7
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BY:-
REVIEWED
BY:-
AUTHORISED
BY :-
Aateka Y
HOD Dr. JP
Sanghshetti
Dr. Abubakar
Salam Bawazir
DATE &
SIGNATURE
26-11-2021
BAY JNS ASB

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