Pir Mehr Ali Shah

Arid Agriculture University, Rawalpindi

Office of the controller of Examinations
Final Exam (Theory)/ Spring 2020 (Paper Duration 48 hours)
To be filled by Teacher

Course No.: ……SS-402………Course Title:……Instrumentation and Lab Techniques………………………

Total Marks:………24.……………………………Date of Exam:……12-08-2020………………………...
Degree: BSc.(Hons.) Agriculture….Semester:………4 th ……… Section:……………………………………
Marks
Q. No. 1 2 3 4 5 6 7 8 9 10 Obtained/
TotalMarks
Marks
Obtained

Total Marks in Words: Twenty Four

Name of the teacher: Dr. Rabia Khalid
Who taught the course: Signature of teacher / Examiner:

To be filled by Student

Registration No.: …18-ARID-2093……… Name:…MUHAMMAD ASIM……………………..

Note: You can extend space as per question requirement.

Q.No.1. Enlist the principle and explain working of spectrophotometer. (06)

Answer:.
Spectrophotometer:- spectrophotometer is an instrument that is used to measure the amount of
light absorbed by the sample.
 Principle of spectrophotometer:-.
Spectrophotometer technique is to measure light intensity as a function of wavelength. It does this
by deflecting the light been into a spectrum of wavelength detecting the intensity with the charge
coupled device and display the result as a graph on detector and then on the display device.
1. In the spectrophotometer a prism grating is used to split the incident beam into
different wavelength.
2. By suitable mechanism wave of specific wavelength can be manipulated to fall on the
test solution.the range of wavelength of the incident light can be as low as 1 to 2nm.
3. The spectrophotometer is used for measuring the absorption spectrum of the
compound that is the absorption of light by solution at each wavelength.
 Working of spectrophotometer
The major sequence of event in spectrophotometry is as follow:-
 The light source shine through a monochromator.
 The output wavelength is selected and aimed at the sample.
  A of the monochromatic light is transmitted through the sample and to the photo detector.
Spectrophotometer in general consists of two devices a spectrometer and a photometer.A
spectrometer is a device that produce typical disperse and my head light.a photo metre indicate the
photoelectric detector that measure the intensity of light.
 Spectrometer. It produce a desired range of wavelength of light. First a collimator (lens)
transmit a straight beam of light(photons) it that pass through monochromator (prism) to
split it into several component wavelength (spectrum). Then a wavelength selector transmit
only a desired wavelength.
 Photometre:- after the desired range of wavelength of light pass through the solution of
sample in cuvettr, the photo metre detect the amount of photon that is absorbed and then
send a signal to galvanometer or digital display.
Beers law : A=£bc
This equation is the heart of spectrometry is called the beer Lambert law. Where A=observance,
£=permittivity constant, b=b is width of cuvet , c is concentration.

Q.No.2. Explain storage of lab chemicals. (06)

Answer:.
Storage of lab chemicals includes:-
1. Storage of Flammable chemicals:-.
 Etremely or highly flammable are stored in a closed steel or thick Bollywood box at
ground level.
 Do not store flammable and oxidizing chemical together.
2. Storage of toxic harmful and irritating chemicals:-
 Potassium cyanide must be kept in a a locked cupboard.
  Stock solution for solid of harmful and irriating chemical should be stored safely in
cupboard not on open shelves.
3. Storage of oxidizing chemicals:-
 Oxidizing chemicals must be stored well away from flammable chemicals and other
chemicals with which they can react dangerously.
4. Storage of corrosive chemicals:-.
 corrosive chemicals should be stored at low level.
 Do not store potassium hydroxide or sodium hydroxide in a bottle having ground glass
stopper because these chemical absorb carbon dioxide from the air and form carbon
cement which can simulate the stopper in the bottle.
5. Storage of explosive chemicals:-.
 Sodium azide can form explosive product one in contact with metal such as copper and
lead.
 perchloric acid when it allowed to dry on woodwork this chemical with will exploit and
cause a fire on impact.
 Diethyl ether and other ethers exposed to air and sunlight ethic and form shocks and
explosive peroxides.
6. Storage of environmentally dangerous chemicals:-.
 Colour code red:-wet infection material like blood body tissue blood bags are dumped in
red colour bin.

Q.No.3. How we can ensure biosafety levels 1 and 2 in a lab? (06)

Answer:.
What is Biosafety levels:- Biosafety level is the level of biocontaminants precaution required to
isolate dangerous biological agent in an enclosed facility.
The level of contaminants range from lowest biosafety level 1 the highest biosafety level 4.
 Why we need biosafety?
 Lab has hazard of processing infection agent.
 Accidental thread to worker and environment.
 To have adhere with safety regulation while dealing with highly infections agent.
  How we can ensure biosafety level 1 and 2 in a lab?
 Biosafety level 1
Microbes not known consistently to cause disease in healthy adults and present minimal potential
hazard to lab and environment. Biosafety level one is suitable for work in wall well characterize
agent not known to consistently cause disease in healthy adult human and minimal potential hazard
to laboratory personal and environment.
 Biosafety level 1 practices and procedure:-
1. Only standard practices required at this level that include Good laboratory practices.
2. Frequently hand washing after removing gloves and before leaving lab.
3. Daw that can be kept close when working. Limit on access to lab space when working.
4. No smoking eating and drinking and also storage of food in the laboratory. Open benchtop
work allowed.
5. Biosafety cabinets not required.
6. Care to to minimise flashes and action that may create aerosol. Daily decontamination.
7. Red bag waste.
8. Decontamination of laboratory waste.
9. Use mechanical pippeting only.sharp precautions include special containers of disposal of
needles and other sharp object.
10. Use of personal protective equipments used during procedure.
11. Use surgical gloves.
12. Open bench top sink for hand washing.
13. Red bag waste is exactly what it sound like-red biohazard autoclave bag with biohazard
waste inside.the biohazard waste could be anything from pipit used in microbiology research
experiment to instrument exposed to human waste from hospitals. For example items goes
into red bag include plastic tubing, pippets and petri dish used in research. Bandages , gauze
nd needle exposed to infection material.
 Biosafety level 2
Biosafety level 2 is similar to biosafety level 1 and is suitable for work involve agent of moderate
potential hazards to personal and environment.SS Dil battery is resist restricted when work is being
conducted.all procedures in which infection aerosol or splash may be created are conducted in
biological safety cabinets or other physical contaminant equipments.
 In biosafety level 2 agent the primary hazard to personal are injection of infectious material.
 Direct contact or exposure.
 Accidental needle stick for per cutaneous exposure by scratch for puncture.potential
infection through exposure to mouth, nose, open cut etc
 Biosafety level 2 practices and procedure:-
1. Limited access to lab when work in progress.
 2. Daily decontamination.
3. Mechanical peptting.
4. Lab coat safety glasses and gloves required.
5. Red bags and sharps container required.
6. Biohazard warning sign posted at entrance to lab with contact information mandatory.
7. Label are equipments including incubator and freezer.
8. Documeted training.
9. Special entry procedures baseline serology for free vaccination may be required.
.

Q.No.4. Explain the calibration of pH meter. What is the importance of calibrating pH meter? (06)
Answer:-.
 Calibration of pH metr
1. Turn on your pH metre:- Before you start to calibrate and use your pH metre you will first
need to turn it on and allow adequate time for the metre to warm up. they should generally
take around 30 minute but check your pH metre operating manual for exact time.
2. Clean the electrode:- Take the electrode out of it storage solution and rinse it with digital
water under an empty waste beaker.once rinsed blot dry with tissue be sure to rinse your
electrode in way speaker that is different from beaker you will be calibrate in. avoid rubbing
the electrode as it has a sensitive membrane around it. if you find the electrode to be
particularly dirty consult your operating manual for recommended cleaning solution.
3. Prepare your buffer:-. you will generally need more than 1 buffer for calibrating a pH metre.
The first it will be natural buffer with PH of 7 and the second should be near the expected
sample pH either at pH 4 or 9.1 buffer with high PH 9.21 are best buffer measuring base
whereas buffer with the low ph 4 are best for measuring acidic sample. once you have
 chosen your buffer allowed them to reach the same temperature as the pH metre same
temperature because pH reading a temperature dependent.. buffer should be kept in a
beaker for no longer than two hours. Discard the buffer when you are finished do not return
it to its original container.
4. Place your electrode in the buffer with a PH value of 7 and begin reading. Press dham
maihar calibrate button to begin reading the pH once your electrode is placed in the
buffer.Allowed the pH reading to stabilize before letting it sit for approximately 1 to 2 minut.
5. Set the pH:- once you stable the readings set the pH metre to the value of buffer pH by
pressing the maihar button a second time.setting the pH metre of once the reading has
stabilized with will allow for more accurate and turned reading.
6. Rinse your electrode with distilled water:- Rinse and dry with lint free tissue in between
buffer.
7. Place electrode in the appropriate buffer for your sample pH 4 and begin readings:-
pratham mayur button to begin reading the pH once your electrode is placed in buffer
8. Set your pH second time;- what's the reading has stabilized press the menu button this is pH
level of your sample.
9. Rinse your electrode:- you can use distilled water to rinse induce lint free tissue to dry
electrode.
10. place your electrode in your sample pH 9.1 and begin the reading.;- once your electrode is
placed in your sample press the measure button and leave for 1 to 2 minut for reading.
11. Set your pH level:- once the reading has tab lies press the mayor button. this is the pH of
your sample.
12. Clean your electrode after use:- rinse your electrode with distilled water and plot dry with
lint free tissue.
------------------------Good Luck------------------------