Lab 1

 Graduated cylinder: measure the volume of liquids.

 Beaker: For Holding liquids.
 Erlenmeyer flask: hold and measure chemical liquid samples, The shape of it allows us to freely
shake the flask without spelling the liquid.
 Burette:
 liver specific amount of liquid
 a burette clamp: hold and secure the burette
 stopcock: (when its horizontal it is closed, when vertical its fully opened)
 The volume marks start with zero at the top to reach the highest at the bottom (as you go
down the numbers increase).
 To determine the volume of liquid that has been transferred from the burette, we take the
difference between final and initial volume
 Funnel: deliver liquids to containers that have small opening (to burette)
 Materials:
 Strong base. [NaOH] = 0.1 M
 Amino acid. [alanine] = 0.1 M
 Distilled water.
 General notes on procedure:
 Fill the burette with NaOH, do not forget to rinse the burette
with NaOH before use to prevent contamination
 10 ml of alanine to the cylinder then to the flask
 Add few drops of concentrated HCl to the alanine in flask in
order to have low initial pH reading.
 measure the pH in the flask by immersion the probe in the
solution, wash the probe using distilled water and dry it using
a clean tissue
 Using the burette, add 1ml of NaOH to the alanine solution.
Shake it well and measure the pH (record the new pH)
 Continue….then table then graph as shown
 You can estimate the buffer regions and the PKa of the
ionizable groups (middle of plateaus)
 PI= (pKa1 + pKa2) /2
Lab 2
 3 glucose samples are generated through serial dilution
 First tube is from stock solution with concentration of 300 mg/dL.
 Second tube we add 1 ml of stock solution to 1 ml of distilled water. So the new
concentration
- So the new concentration is: C1V1=C2V2 -> 300x1=C2x2 -> C2=300/2 -> C2= 150 mg/dL
- DF is 2/1
 Third tube: we add 1 ml of liquid in second tube “1 ml of concentration 150mg/dL” to 1 ml
of distilled water -> C1V1=C2V2 -> 150x1=C2x2 -> C2=150/2 -> C2= 75 mg/dL. So now the
final dilution factor is 4 times.
 The fourth test tube will have the unknown sample X
 Measure their absorbance using spectrophotometer and then draw the standard curve
(linear curve)
 Note: we can use the equation 𝐂𝟐 = ( 𝐀𝟐/ 𝐀𝟏) 𝐱𝐂1
 Equipments:
1. Glass pipette 2. Fixed volume micropipettes with 2 with different sizes.
3. Tips for the pipettes. 4. Test tubes.
 Procedure:
 Serial dilution: perform serial dilution to get multiple glucose samples with known
concentrations (300, 150, 75 mg/dL)
 We use the Glass pipette to transfer solutions during dilution
 Now bring 6 empty tubes. We will label them as:
 1- Blank. 2- 300mg/dl 3- 150mg/dl 4- 75mg/dl 5- Sample A 6- Sample B
 We use the Fixed Volume Micropipette to add 1 ml of glucose reagent on each tube
Note: changing the tip with each use to prevent contamination.
 We will use another Fixed Volume Micropipette with different size to add 10 microliters
(0.01 ml) of each concentration to the 6 tubes.
 wait for 15 minutes at room temperature for the reaction to happen. At higher
temperatures (>35 C), the reaction needs less time (10 minutes).
 waited for: Only 8 minutes: the glucose level will be underestimated “less that the real
level”.
 Measure the absorbance:
- Adjust the wavelength to the desired wavelength (by pressing GOTOλ and setting λmax
at 500nm, visible light)
- Zero Adjustment: put the Blank solution “the reagent only” in a plastic cuvette and put it
in the sample compartment in the spectrophotometer then, press Zero button.
- After that you can measure the absorbance of all the samples.
- Construct the concentration-absorbance graph: find the concentration that corresponds
with the unknown sample absorbance
- the color intensity is stable for 30 minutes
- the normal glucose level in serum is 75-115 mg/dL
Lab 3
 We need:
1. Biuret reagent
2. Protein standard with known conc.
3. Samples with unknown concentrations [A and B]
4. Fixed volume micropipette and Glass pipette
5. 4 test tubes labeled as : [Blank, protein standard, A, B
 PROCEDURE:
 Using the glass pipette add 5ml of the reagent to each test tube.
 Use the fixed volume micropipette to take 100μl of protein standard.
 Add it to the protein standard tube and shake it well [ note the change in the color the
violet color starts to appear]
 Use the micropipette to take 100μl of sample A and add it to test tube A then shake it well
 Repeat the same step for sample B
 Wait for 10 minutes to get complete reaction

Note: This is non-enzymatic colorimetric reaction

 Measure the absorbance for standard and samples using spectrophotometer

 Adjust the wavelength to the desired one [ for this experiment the wavelength required is
545nm] by pressing the button “go to ƛ “
 Transfer the blank solution which contains only the reagent to the plastic cuvette Then get
the cuvette into the sample compartment in the spectrophotometer
 Press ”zero” so [zero adjustment with the blank reagent ]
 To measure the absorbance of the protein standard solution transfer the solution from the
test tube to the cuvette then get it into the sample compartment to measure the
absorbance ]
 Cx = Ax.Cs/As