BIO-CHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and

Purpose
maintenance of the UV – Visible Spectrophotometer

This instrument is used to measure how much light of a

Scope
given wavelength is absorbed by a liquid sample.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR

UV – VISIBLE SPECTROPHOTOMETER
 Open panel door and make sure cuvette holders are empty, and then close the
panel door.
 Flip open the display unit and Turn spectrophotometer “ON” by flipping the
yellow switch on the side of the machine. The machine will automatically
initialize..
 Hold the cuvette from the top to prevent tampering with the measurements, and
wipe the sides with a lab tissue.
 Open panel door and place the cuvettes with blank solution in the cuvette holders.
When the Auto Zero is complete, open the panel door and remove the front
cuvette.
  Do not replace cuvette in rear holder.
 Using the same cuvette style, fill an empty cuvette with about 2-ml of the sample.
 Clean the cuvette with a lab tissue.
 Place in front cuvette holder, using the appropriate orientation and close the panel
door.
 Press START to take a reading.
 Select spectrum from the drop down menu
 Press save as and give name and save.
 Open panel door and remove test sample from front cuvette holder.
 To test additional samples: Place cuvettes in front holder and press start for a
reading.
 Press RETURN to bring you back to step Note: This will erase your old data.
 Remove cuvettes remaining in holders.
 Then click on the Disconnect button near the bottom of the screen.
 Exit from the UV-Probe software.
 Push the mode button on the UV-VIS instrument keypad.
 Flip power switch located on the side, to turn off the machine.
PRECAUTIONS TO BE FOLLOWED
N/A
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and

Purpose
maintenance of the Soxhlet apparatus

This Standard Operating Procedure (SOP) applies to the

staff and students using the Soxhlet apparattus in the Bio
Scope
chemistry and Bioinstrumentatiom laboratory, Vinayaka
missions’ Research foundation

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR SOXHLET APPARATTUS

 Assemble the apparatus.

 Fill the round bottom flask with solvent (n-Hexane).
 Put the thimble containing sample into extraction tube.
 Attach the extraction tube with flask containing solvent.
 Attach a condenser unit with the extraction tube and run the water.
 Fix the soxhlet apparatus on hot plate and heat the flask containing solvent.
 The solvent starts to evaporate and falls in the extraction tube after condensing.
 Continue this process till all the fat is extracted.
 Discontinue the process and take out the thimble.
 Again attach the extraction tube with flask containing solvent along with fat and
condenser unit.
 Again heat the flask to recover the solvent.
 Discontinue the process and clean the extraction tube and thimble.
PRECAUTIONS TO BE FOLLOWED
 Always perform Soxhlet extractions in a fume hood (with sash down) to avoid
inhalation of hazardous solvents and to contain potential fire
 Personal Protective Equipment – Eye protection: required – Hand protection:
required
 Nitrile gloves are recommended
 Insulated gloves used for contact with hot glassware – Protective Clothing: lab
coats, closed toe shoes and long pants should be worn
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and

Purpose
maintenance of the Conductivity Meter

This standard operating procedure provides the laboratory

Scope person with guidance on the procedure for determining
conductivity in Liquid and water samples.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR CONDUCTIVITY METER

 Turn on the meter.

 Rinse the probe with DI water.
 Dip probe into sample making sure there are no air bubbles trapped on the slot of
the probe.
 Stir the probe gently in the sample to create a homogenous sample, allowing a few
seconds for the temperature reading to approach the solution temperature
 Take reading (when the reading is stable ‘READY’ will display on the left-middle
of the LCD.
PRECAUTIONS TO BE FOLLOWED
Do not use or install the this equipment in any manner other than which is
 specified
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and

Purpose
maintenance of the Centrifuge

This Standard Operating Procedure (SOP) applies to the

staff and students using the Centrifuge in the
Scope
Biochemistry and Bioinstrumentation Lab, Vinayaka
missions’ Research foundation

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR CENTRIFUGE

 Press the start/stop button and slowly increase the rpm to the desired speed using
the dial
 Once a run is complete, make sure the rotor has COMPLETELY STOPPED
before opening the centrifuge lid by depressing the red stop/start button.
 Remove sample vials.
 Remember to return the rpm dial back to zero after finishing.
PRECAUTIONS TO BE FOLLOWED
 Proper handling of the instrument
 Ensure level and stability
 Balance centrifuge tubes equally
 Ensure use of rubber cushion for glass tubes
 Bring speed Knob to off and increase the speed gradually.
 Do not open the lid in between the centrifugation cycle
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and

Purpose
maintenance of the Magnetic Stirrer

This Standard Operating Procedure (SOP) applies to the

staff and students using the Magnetic Stirrer in the
Scope
Biochemistry and Bioinstrumentation laboratory,
Vinayaka missions’ Research foundation

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR MAGNETIC STIRRER

 Place the magnetic stirrer on a stable well-levelled surface.

 Place the stir bar at the bottom of a glass container.
 Fill the glass container with the liquid to be stirred.
 Plug the mains cable into a suitably earthed socket.
 Check that the speed control knob is completely turned anticlockwise.
 Place the glass container on the centre of the magnetic stirrer.
 Press the On/Off switch to turn the magnetic stirrer On. The switch will light
green.
 Adjust the speed control knob to a low stirring rate.
 Continue to adjust the speed control knob until the desired stirring speed is
 achieved.
 Wait until the liquid is properly mixed.
 Completely turn the speed control knob anticlockwise.
 Press the On/Off switch to turn the magnetic stirrer Off
 Manipulate another stir bar from the outside of the glass container to remove the
immersed stir bar
PRECAUTIONS TO BE FOLLOWED
 Thoroughly wash the stir bar with distilled water after each application.
 Store stir bars in pairs to maintain their magnetic strength and increase their life
span.
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81-BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and

Purpose
maintenance of the pH meter
This SOP describes how to use and maintain a pH meter.
The pH meter is intended for accurate measurement of the
pH of buffer solution used in the Biochemistry and
Scope
Bioinstrumentation laboratory.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR pH METER

 Before use, rinse the electrode with deionized water and blot dry with a soft, clean
paper towel.
 Transfer the electrode to the test solution.
 Compensate for the temperature if necessary.
 Record the pH when the reading is stable (5–20 seconds after insertion of the
electrode into the solution)
 Rinse the electrode with deionized water and store according to the manufacturer’s
instructions.
 Cleaning and maintenance
 Clean the pH meter with a soft, clean, damp paper towel after use. No solvents
should be used.
 Replace the electrode filling solution on a regular basis, according to the
manufacturer’s instructions. Record in the logbook (Adjust the speed control knob
to a low stirring rate.
PRECAUTIONS TO BE FOLLOWED
 Thoroughly wash the stir bar with distilled water after each application.
 Store stir bars in pairs to maintain their magnetic strength and increase their life
span.
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and

Purpose
maintenance of the Colorimeter
This SOP describes how to use and maintain a
Colorimeter. The ColoriSmeter is intended for accurate
measurement of the concentration of the given sample
Scope used in the Biochemistry and Bioinstrumentation
laboratory.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR COLORIMETER

 Switch on the instrument at least 5 minutes before use to allow it to stabilize.

 Select the most appropriate filter for the analysis and insert it in the light path or
dial it in with the selector .
 Place the reagent blank solution (or water) in the cuvette and zero the instrument .
Make sure the clear faces of the cuvette are in the light path
 Place the sample in the colorimeter and read the absorbance of the solution. If the
absorbance is "over range" (usually > 2.0) then the sample must be diluted to yield
a value within the limits of the instrument.
 At intervals, recheck the reagent blank to ensure that there is no drift in the zero
 value.

PRECAUTIONS TO BE FOLLOWED

 The machine must be re-zeroed , if a new filter is chosen.

 The sample cuvettes must be at least two-thirds full.

 The outside of the cuvette must be dry and clean.

RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and
Purpose maintenance of the Hematology Auto Analyzer

This SOP describes how to use and maintain a

Hematology Auto Analyzer. This performs automated
blood counts and requires no manual operations for
aspirating blood, dilutions, measuring, calculations, print-
Scope
outs and computer transfer used in the Biochemistry
laboratory.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR HEMATOLOGY AUTO

ANALYZER
Specimen requirements:

 About 2-3 ml of venous blood collected into EDTA tubes. Specimens should be
transported at room temperature 18 - 26ºC and can be store in the refrigerator of 2
- 8ºC up to 6 hours. If stored in a refrigerator, samples should be returned to room
temperature, for approximately 30 minutes, before analysis.
PROCEDURE:

1.Check operation of the machine, ensuring it is clean and that all required supplies are
present in sufficient quantities.
2. Switch the instrument on by pressing the ON/OFF switch, located on the back of the
instrument.
3. The instrument performs an initialization phase for the internal electronics. Please
wait.
4. Once the initialization phase is complete, the analyzer will automatically run a startup
cycle.
5. If it does not automatically run a startup cycle after the initialization phase is
completed, press "Startup" button in the "Status" area to initiate a startup cycle.
6. Then, the instrument will perform a blank cycle for a reference blank count (an
analysis cycle based on reagents without any blood sample).
7. Check and verify that the reference blank counts do not exceed the following
parameter limits: WBC < 0.3, RBC < 0.02, HGB < 0.3, PLT < 10 then: Press "OK"
button to validate blank results. 8. Perform quality control analysis on 3 levels of control
blood material (low, normal and high) to verify that the instrument is performing within
the specified ranges of the quality control material.
9. Entering patient ID, sample ID, Patient name, etc
10. Follow the indications displayed in the "Sample analysis" dialog box to run the
analysis. a. Mix the sample gently and thoroughly. b. Remove the cap from the sample
tube. c. Place the sample beneath the sampling needle. d. Raise up the tube so that the
sampling needle lowers into the blood and press the manual sample bar. e. The analysis
cycle will begin.
11. When the analysis is completed, the "Sample analysis" dialog box is closed and
results are displayed in the "Result display" menu for print out.
PRECAUTIONS TO BE FOLLOWED
Criteria for rejection haematology specimens 1. When the identification is missing
/inadequate. 2. Insufficient quantity. 3. Inappropriate container 4.Inappropriate
transport/storage. 5. Unknown duration of delay. 6. Clotted sample.
Equipment
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and
Purpose maintenance of the Biochemistry Analyzer

This SOP describes how to use and maintain a

Biochemistry Analyzer. This performs automated blood
counts and requires no manual operations for aspirating
Scope blood, dilutions, measuring, calculations, print-outs and
computer transfer used in the Biochemistry laboratory.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR BIOCHEMISTRY ANALYZER

PROCEDURE:

1. Check the Analyzer is attached with Electric supply.

2.Switch on the Analyzer
3. Press Aspiration key to take water, wash it thrice.
4.Keep the blank and press the aspiration button
5. Keep the standard solution and press the aspiration bottom.
6. Keep the test sample and press the aspiration button and record the reading
7.wash using water
8.Switch the Analyzer
PRECAUTIONS TO BE FOLLOWED
 Keep away from direct Sunlight
 Keep in dry and cool space
RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and
Purpose maintenance of the Turbidity Meter.
Clarity of water is extremely important in products
destined for human consumption. Suspension free water is also
considered necessary for industries manufacturing beverage,
food products etc. Similarly, minimum limits of turbidity are
required to be observed in drinking water & sewage disposals.

Digital Turbidity Meter and Nephelometer are ideal

Scope instruments for measurement of suspensions. They consist of a
light source focused on a test tube containing the sample
solution under test. The light, reflected at right angle to the
focused light by the suspension in the solution, is detected by the
photo electric detector, amplified and displayed on a 3½ digit
LED display.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR TURBIDITY METER

PROCEDURE:

1. Fill a clean container with at least 50 ml. of sample water and cover. Set sample aside
to allow sample to equilibrate to air temperature and let gas escape. Avoid contaminants
and analyze as soon as possible.

2) Rinse 2 empty turbidity tubes and caps with a portion of sample. Shake out excess
water.

3) Fill both turbidity tubes to the neck by carefully pouring the sample down the side of
the tube. This will prevent air bubbles from forming. (Similar to pouring soda or beer
into a glass while trying to avoid foam on the top).

4) Cap the tubes and wipe dry with a clean lint free tissue.

5) Carefully invert the turbidity tube twice (do not create air bubbles) just before
inserting the tube into the meter,

6) Open the meter lid. Align the indexing arrow on the tube with the indexing arrow on
the meter. Insert the turbidity tube into the chamber.

7) Close the lid. Push the READ button.The turbidimeter in NTU units will be displayed
within 5 seconds.

8) Record the results from each tube. Take a third reading from a new sample if the
first two readings are significantly different.

9) The meter will turn off automatically one minute after the last button push. To turn
the meter OFF manually, hold the READ button down for at least2 seconds. Release the
button when OFF is displayed.

PRECAUTIONS TO BE FOLLOWED
 ER1 (ERROR 1) will be displayed if the battery is low.
 ER2 will be displayed when the measured turbidity is very low.
 ER3 will be displayed when the light source (bulb) has burned out or if the
turbidity tube is misaligned.
  BAT will be displayed when the battery is getting low. The readings are
reliable but the battery needs to be replaced as soon as possible. Carry extra
batteries with the field kit.

RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
equipment
 Maintenance Record

HoD
 17BTCC81 -BIOCHEMISTRY LABORATORY

STANDARD OPERATING PROCEDURE

Name of the Lab./facility BIOCHEMISTRY AND BIO INSTRUMENTATION LAB

To describe the procedure for the operation and
Purpose maintenance of the Turbidity Meter.

Entry-level centrifuges that are powerful and easy-to-use.

Scope They are small enough so that each workstation can be
equipped with a personal« centrifuge.

It is the responsibility of Lab – In-charge of the lab to

train Lab Assistant and students on this procedure and to
ensure adherence to this procedure.
Responsibility It is the responsibility of the students/technicians to
follow the SOP as described and to inform the Lab In-
Charge about any deviations or problems that may occur
while performing the procedure.

STANDARD OPERATING PROCEDURE FOR MINI SPIN

PROCEDURE:
PRECAUTIONS TO BE FOLLOWED

RECORD TO BE MAINTAINED
 Laboratory Manual containing the experiments that can be performed with the
 equipment
 Maintenance Record

HoD