Anika Wijesuriya (42213017) 27/02/2024 Lab ID: 32322

Abstract
The acid/base buffer capacities of a phosphate system were tested with Hydrochloric Acid
(HCl) and Sodium Hydroxide (NaOH) to imitate the buffering capacities of human blood. A
drop dispenser was used to titrate both the acid and base. Another titration was used to
determine the amount of “Restoration Solution” needed to stabilise the pH of a blood
sample. It was concluded that the acid buffer capacity of blood is greater than its base
buffer capacity.

Introduction
The blood system maintains an acid-base balance, where the pH ranges from 7.35 to 7.45,
and is essential for the functionality of the body, one example being the oxygenation of
blood1. If the blood system is too acidic, when the alkalinity of blood overly reduces, then
acidosis occurs2, conversely if it is too basic, alkalosis occurs. Disorders involving acidosis
and alkalosis include metabolic acidosis, metabolic alkalosis, respiratory acidosis, and
respiratory alkalosis.1 The human body has several ways of preserving this balance, which
makes the blood system a “buffer mixture”2, where blood can absorb large amounts of acid
or base without a large change in pH.

In this experiment, the blood buffer system was studied using a phosphate buffer system to
determine how acids and bases reacted in a buffered environment. The hypothesis was that
the acid buffer capacity would be much larger than its base capacity. A drop dispenser was
used to titrate HCl and NaOH respectively. The volumes for both HCl and NaOH were then
used to calculate the buffer capacity. With this knowledge, a sample of patient blood, which
was slightly acidic, was treated with restoration solution such that the pH stabilised.

Experimental
The “Microlab” program was set up to record the pH of the solution as each drop of
acid/base was added. The pH itself was measured by a pH probe, which was placed in the
solution. The drop dispenser was calibrated by letting around 100 drops of deionized water
fall into a beaker which was then weighed. The volume of each drop was then found by
dividing the volume by the number of drops, which was typed into the Microlab program.

The buffer was made by adding 1.0 M NaOH to 35.8 mL of 0.5 M H2PO4 until a pH of 7.45 was
reached. The solution was then transferred to a 100 mL volumetric flask and diluted to
volume with pre-boiled deionized water. 25 ml of the buffer was pipetted into a 250 mL
beaker, while 0.1 M HCl was placed into the drop dispenser and allowed to flow until the
pH reached 6.45. The Microlab program then printed out a graph. This was repeated with
0.1 M NaOH, and with the “restoration solution” (0.2 M of Na2HPO4). The data collected
from the restoration solution titration was then used to recover the pH of a patient blood
sample back to 7.45.
Results
Table 1: Acid and Base Buffer Capacities

Acid Buffer Capacity (M) 0.08780 M

Base Buffer Capacity (M) 0.03328 M

Discussion

The results of this experiment confirm the hypothesis that the acid buffer capacity (0.08780
M) was larger than the base buffer capacity (0.03328 M). The phosphate buffer system
(H2PO4-/HPO42-) functions like the carbonic buffer system (H2CO3/HCO3-) found in humans.
The phosphoric acid replaces carbonic acid in this model and reacts with the OH - ions of the
NaOH, creating its conjugate base (HPO42-), which would react with the H3O+ ions found in
HCl and re-create H2PO4-. This is how the buffer system maintains a steady pH level.
The addition of HCl models the production of acid due to metabolic activities. One example
is the build up of lactic acid when exercising. After the lactic acid is formed, it is cleared
into the bloodstream to be metabolised, where the buffer system will stabilise the pH.
Metabolic processes produce more acid than base which explains why the human
bloodstream has a higher acid buffer than base buffer capacity.

Acidosis was the main issue with the patient sample blood. To rectify this, a titration using
the Restoration Solution on the trial blood was required and produced a graph. The initial
pH of the sample was 7.05. Based on the graph, 25 mL of restoration solution was required
to save the “patient”. The calculated amount was correct, and the trial was successful.

The pH probe reading different pH levels was an issue. It originally read 7.45 but when
pipetted to another beaker it was 7.55, which is why the graphs for HCl and NaOH both start
at 7.55. A possible explanation for this could be that the volumetric flask was not rinsed
well and held traces of some kind of basic solution that caused the pH to increase. In future,
maybe adding a small amount of HCl and NaOH to neutralise the remnants of the flask and
better rinsing would improve the accuracy of the experiment.

Conclusion
The results of the experiment confirm the hypothesis that the acid buffer capacity of the
blood buffer system is larger than its base capacity. The objectives of this study have been
met, and further data has proved that 25 mL of restoration solution was required to stabilise
the given sample of patient blood.

Bibliography
1
Hopkins, E.; Sharma, S.; Sanvictores, T. Physiology, Acid Base Balance. National Library of
Medicine https://www.ncbi.nlm.nih.gov/books/NBK507807/.
2
LEVY RL, ROWNTREE LG. A STUDY OF THE BUFFER VALUE OF THE BLOOD. Arch Intern
Med (Chic). 1916;XVII(4):525–539. doi:10.1001/archinte.1916.00080100069007

2
Appendix
D1) SEE EDF

D2) *Volume was found through the graphs*

( )( )
𝐴𝑐𝑖𝑑 𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 =
( . )( . )
𝐴. 𝐵. 𝐶 =
𝐴. 𝐵. 𝐶 = 0.08780 𝑀

D3) *Volume was found through the graphs*

( )( )
𝐵𝑎𝑠𝑒 𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 =
( . )( . )
𝐵. 𝐵. 𝐶 =
𝐵. 𝐵. 𝐶 = 0.03328 𝑀

D4) Yes, the acid buffer capacity is greater than the base buffer capacity.

D5) From the graph, 25 mL of restoration solution was needed to restore the pH to 7.45
from the original pH of the solution (7.05).

D6) Since the sample was already acidic, the restoration solution probably had the
phosphate ion (HPO42-) in order to neutralise the H3O+ ions.

D7) Volume of restoration solution needed to restore the pH to 7.45 is 25 mL for 25 mL of

patient trial blood. The question asks for the volume needed for 7L.
1000 𝑚𝐿
7𝐿 × = 7000 𝑚𝐿
1𝐿
The ratio of trial blood to patient blood is 25 ml:7000 ml.
7000 𝑚𝐿
25 𝑚𝐿 𝑟𝑒𝑠𝑡𝑜𝑟𝑎𝑡𝑖𝑜𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 × = 7000 𝑚𝐿 𝑟𝑒𝑠𝑡𝑜𝑟𝑎𝑡𝑖𝑜𝑛 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
25 𝑚𝐿
Therefore 7000 mL of restoration solution is needed.

D8) Looking at the graph it took 4.4 mL of restoration solution to restore the pH from 7.40
to 7.45. It took 12.2 mL from 7.30, 18.2 mL from 7.20, 23 mL from 7.10, and 26.8 mL from
7.00. Each volume is then multiplied by 4 to get approximately 100 mL of buffer. This was
then multiplied by the kg value. ex) (23 * 4) = 92 mL, (92 * 50 kg) = 4600 mL

pH Volume of Restoration Solution (mL)

7.40 880 968 1056 1144 1232 1320 1408

7.30 2440 2684 2928 3172 3416 3660 3904

7.20 3640 4004 4368 4732 5096 5460 5824

7.10 4600 5060 5520 5980 6440 6900 7360

5360 5896 6432 6932 7504 8040 8576

7.00

50 55 60 65 70 75 80 kg