The pH in Blood: Experimental Model of a Blood Buffer and its Buffer Capacities

Bashar Adamat, CHEM 123, March.6th, 2024 Lab ID: 51207

Abstract

This experiment determines if the acid buffer capacity of a phosphate buffer will exceed the
base buffer capacity and if the phosphate buffer effectively models the human blood buffer. Titrations
with NaOH and HCl were performed, and the patient trial blood was titrated against the restoration
solution was done to find the volume of the restoration solution needed to save the patient from
acidosis. 1.0 M NaOH and 0.5 M H2PO4 - was used to create a phosphate buffer. The acid buffer
capacity was higher than the base buffer capacity and the phosphate buffer effectively modelled the
human blood buffer.

Introduction

For humans to maintain normal cell function, a blood pH of 7.35 to 7.45 is needed (Hughes et
al., 2021). Otherwise, blood pH that becomes excessively acidic (acidosis) or excessively alkaline
(alkalosis) will result in an immediate coma or death (Rosival, 2011). To prevent these conditions, a
buffer system, a solution that resists large changes in pH, is needed for a stable blood pH. Researchers
have investigated the impact of increasing the acidity or alkalinity of buffers in human systems such as
the hydrogen carbonic acid-bicarbonate buffer in human blood (Thomas, 2022). Since metabolic
processes lower pH (Thomas, 2022), an observation found through previous research, it is hypothesized
that the acid buffer capacity will be higher than the base buffer capacity for a phosphate buffer. My
hypothesis is that 25 mL of restoration liquid will be needed to bring the pH of the blood to 7.45. This
experiment analyzes how the addition of a strong acid and strong base can be used to determine
whether the acid buffer capacity will exceed the base buffer capacity of a model human blood buffer.
This experiment was conducted using a phosphate buffer and titrations with HCl and NaOH to find the
acid and base buffer capacities. The phosphate buffer was used as a model human blood buffer since
the solution can withstand decomposition as opposed to the carbonate buffer (Thomas, 2022). A patient
trial procedure was also conducted to find the amount of restoration solution needed to rescue patients
from acidosis.

Experimental Section

To prepare the phosphate buffer, 36.0mL of 0.5 M H2PO4 - and 12 mL of 1.0M of NaOH were
mixed. The pH probe was first calibrated by placing the probe in a buffer of pH 7 and then used to
monitor the pH of the buffer. 1.0M NaOH was added dropwise to 0.5 M H2PO4 - until the pH reached
7.45. Afterwards, preboiled deionized water was added to the buffer solution until the volume was 100
mL. The Microlab drop counter and the dispenser were calibrated for the titrations. Afterwards, the acid
buffer capacity was found by dispensing the 0.1 M HCl into 25.00 mL of the buffer solution until the pH
dropped by 1 unit. To find the base buffer capacity, 0.1 M NaOH was dispensed into 25.00 mL of the
buffer solution until the pH increased by 1 unit. The recorded volumes of the NaOH and HCl dispensed
were used for buffer capacity calculations. Restoration solution was added to 25.00 mL of the patient’s
blood dropwise until a pH of 7.45 was restored to find the amount of restoration solution needed to
recover a patient from acidosis.

Results

Table 1: Acid and base buffer capacities of the phosphate buffer

Acid buffer capacity (mol of HCL/L of buffer) 9.28 ∗ 10^-2 𝑚𝑜𝑙/𝐿 H3O+
Base buffer capacity (mol of NaOH/L of buffer) 2.98 ∗ 10^-2 mol/L OH-

Discussion

The results indicate that the acid buffer capacity (9.28 ∗ 10^-2 𝑚𝑜𝑙/𝐿) was greater than the base
buffer capacity (2.98 ∗ 10^-2 𝑚𝑜𝑙/𝐿), an observation supportive of the hypothesis. A phosphate buffer
was prepared in this experiment to model the response of a human blood buffer when acids and bases
are added to the bloodstream. The phosphate buffer was like the carbonate buffer (human blood
buffer) in that its acidic component (H2PO4 -) resembled similar behavior to the H2CO3 component of
the carbonate-bicarbonate buffer and its alkaline component (H2PO4 -) behaved similarly to the HCO3 -
component of the carbonate-bicarbonate buffer. Adding HCl to the buffer represents acid production
from metabolic processes. Metabolic processes usually produce acidic products because CO2 acidifies
the blood, meaning that there is more resistance to acid than base, another observation supportive of
the results and hypothesis. To establish an emergency room procedure to save the patient from
acidosis, restoration solution was titrated dropwise against the patient blood, as shown in the graph of
restoration liquid blood pH vs volume. The volume of restoration solution needed to save the patient
from acidosis in a simulated emergency room procedure was predicted using the graph of restoration
liquid blood pH vs volume. Since the sample had an initial pH of 7.00, 21 mL of restoration solution was
added to the patient blood sample until the pH was restored to 7.45. That volume restored the pH to
7.45, thereby saving the patient from acidosis. In contrast, there are a few experimental errors that
could have happened. For instance, the speed of the magnetic stirrer was not constant for each titration
trial, leading to less accurate pH readings. It is recommended to have a consistent speed for the
magnetic stirrer for more Furthermore, the drop counter might have not been fully rinsed after each
titration trial, leading to contamination of the titrant used. In the future, the drop counter should be
completely rinsed with deionized water to ensure no traces of previously used substances. Another
experimental error could be the rapid drop counting when adding strong acid or base during the
titration trials, leading to excessive addition of the titrant and less accurate pH readings and buffer
capacity calculations. Conclusion The phosphate buffer had a higher acid buffer capacity (9.28 ∗ 10^-2
𝑚𝑜𝑙/𝐿) than the base buffer capacity (2.98 ∗ 10^-2 𝑚𝑜𝑙/𝐿) and resembled similar behavior to human
blood buffers in response to the addition of a strong acid or base, meaning that the phosphate buffer
can effectively model the human blood buffer, as hypothesized.

Bibliography

Hughes, J. M.; Vilchiz, V. H.; Lee, C. An Easy Approach to Understanding Acid-Base Balance in a Blood
Buffer System.
The American Biology Teacher 2021, 83 (8), 526–531. Rosival , V. ( 2011 ).

Dangers of very low blood pH . Indian Journal of Critical Care Medicine , 15 , 194

Thomas, A. Chem 123 Experiment 12 Overview; University of British Columbia: Vancouver, BC, 2022; p
1- 9.

Appendix

Graph 1 NaOH experiment pH vs Volume: The pH changes of solution when 36.0 mL of 0.5 M H2PO4 - is
titrated against 0.1 M NaOH. The pH increases as the volume of NaOH increases.

Graph 2 HCL experiment pH vs volume: A HCl titration curve where 36.0 mL 0.5 M H2PO4 - is titrated
against 0.1M HCl. The pH decreases as the volume of HCl increases.

Graph 3 restoration solution blood pH vs volume: A graph of the patient trial blood experimental
titration. The pH increases as the volume of restoration solution is added.

Reminder for TA: all the graphs are with my lab partner Lucca

D8: Volume(mL) of Restoration Solution needed to save patients. The volumes of restoration solution
vary by blood pH and body mass(kg).

pH

7.40 404 444 485 525 566 606 646

7.30 1284 1412 1541 1669 1798 1926 2054
7.20 1944 2138 2333 2527 2722 2916 3110
7.10 2464 2710 2957 3203 3450 3696 3942
7.05 3044 3348 3653 3957 4262 4566 4870
Weight 50 55 60 65 70 75 80
D1(Appears in calculations section of the EDF): The volume of the weak acid needed is 35.9 mL.

D2:

𝐴𝑐𝑖𝑑 𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = (𝑀𝑜𝑙𝑎𝑟𝑖𝑡𝑦 𝑜𝑓 𝑎𝑐𝑖𝑑)(𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑎𝑐𝑖𝑑 𝑢𝑠𝑒𝑑(𝐿))/𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑏𝑢𝑓𝑓𝑒𝑟(𝐿) =

(0.100𝑀)(0.0232 𝐿)/0.02500 𝐿 = 9.28 *10^-2 𝑚𝑜𝑙/L

𝐴𝑐𝑖𝑑 𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = 9.28 *10^-2 𝑚𝑜𝑙/L

Base 𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = (𝑀𝑜𝑙𝑎𝑟𝑖𝑡𝑦 𝑜𝑓 Base)(𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 Base 𝑢𝑠𝑒𝑑(𝐿))/𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑏𝑢𝑓𝑓𝑒𝑟(𝐿) =

(0.100𝑀)(0.00745 𝐿)/0.02500 𝐿 = 2.98 *10^-2 𝑚𝑜𝑙/L

Base 𝐵𝑢𝑓𝑓𝑒𝑟 𝐶𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = 2.98 *10^-2 𝑚𝑜𝑙/L

D4: The acid buffer capacity is greater than the base buffer capacity.

D5: The volume of restoration solution needed to reach a final pH of 7.45 was 15.22 mL.
D6: The restoration solution is comprised of the hydrogen phosphate (HPO4 2-) component of the
phosphate buffer since the addition of the restoration solution increased the pH of the buffer solution.

D7: Volume Restoration solution(mL) needed to restore pH in 7.0 L of blood = (volume needed for 25.00
mL) (volume of patient blood/volume of sample patient blood) = (21 mL) (7.0 L/0.0250 L) = 5.88 L

D8: 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑐𝑒𝑟𝑡𝑎𝑖𝑛 𝑏𝑜𝑑𝑦 𝑚𝑎𝑠𝑠 = (𝑣𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 𝑥 𝑝𝐻 𝑎𝑛𝑑 7.45 𝑝𝐻) (𝑚𝑎𝑠𝑠 𝑜𝑓 𝑝𝑎𝑡𝑖𝑒𝑛𝑡) (4)

For pH = 7.40

𝑉𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 7.40 𝑎𝑛𝑑 7.45 𝑝𝐻 = 2.02 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 50 𝑘𝑔 = (2.02 𝑚𝐿)(50 𝑘𝑔)(4) = 404 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 55 𝑘𝑔 = (2.02𝑚𝐿)(55𝑘𝑔)(4) = 444 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 60 𝑘𝑔 = (2.02 𝑚𝐿)(60 𝑘𝑔)(4) = 485 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 65 𝑘𝑔 = (2.02 𝑚𝐿)(65 𝑘𝑔)(4) = 525 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 70 𝑘𝑔 = (2.02 𝑚𝐿)(70 𝑘𝑔)(4) = 566 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 75 𝑘𝑔 = (2.02 𝑚𝐿)(75 𝑘𝑔)(4) = 606 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 80 𝑘𝑔 = (2.02 𝑚𝐿)(80𝑘𝑔)(4) = 646 𝑚𝐿

For pH = 7.30

𝑉𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 7.30 𝑎𝑛𝑑 7.45 𝑝𝐻 = 6.42 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 50 𝑘𝑔 = (6.42 𝑚𝐿)(50𝑘𝑔)(4) = 1284 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 55 𝑘𝑔 = (6.42 𝑚𝐿)(55 𝑘𝑔)(4) = 1412 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 60 𝑘𝑔 = (6.42 𝑚𝐿)(60 𝑘𝑔)(4) = 1541 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 65 𝑘𝑔 = (6.42 𝑚𝐿)(65 𝑘𝑔)(4) = 1669 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 70 𝑘𝑔 = (6.42 𝑚𝐿)(70 𝑘𝑔)(4) = 1798 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 75 𝑘𝑔 = (6.42 𝑚𝐿)(75 𝑘𝑔)(4) = 1926 𝑚L

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 80 𝑘𝑔 = (6.42 𝑚𝐿)(80 𝑘𝑔)(4) = 2054 𝑚L

For pH = 7.20

𝑉𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 7.20 𝑎𝑛𝑑 7.45 𝑝𝐻 = 9.72 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 50 𝑘𝑔 = (9.72𝑚𝐿)(50𝑘𝑔)(4) = 1944 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 55 𝑘𝑔 = (9.72 𝑚𝐿)(55 𝑘𝑔)(4) = 2138 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 60 𝑘𝑔 = (9.72 𝑚𝐿)(60 𝑘𝑔)(4) = 2333 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 65 𝑘𝑔 = (9.72 𝑚𝐿)(65 𝑘𝑔)(4) = 2527 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 70 𝑘𝑔 = (9.72 𝑚𝐿)(70 𝑘𝑔)(4) = 2722 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 75 𝑘𝑔 = (9.72 𝑚𝐿)(75 𝑘𝑔)(4) = 2916 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 80 𝑘𝑔 = (9.72 𝑚𝐿)(80 𝑘𝑔)(4) = 3110 𝑚𝐿

For pH = 7.10

𝑉𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 7.10 𝑎𝑛𝑑 7.45 𝑝𝐻 = 12.32 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 50 𝑘𝑔 = (12.32 𝑚𝐿)(50𝑘𝑔)(4) = 2464 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 55 𝑘𝑔 = (12.32 𝑚𝐿)(55 𝑘𝑔)(4) = 2710 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 60 𝑘𝑔 = (12.32 𝑘𝑔)(60 𝑘𝑔)(4) = 2957 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 65 𝑘𝑔 = (12.32 𝑚𝐿)(65 𝑘𝑔)(4) = 3203 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 70 𝑘𝑔 = (12.32 𝑚𝐿)(70 𝑘𝑔)(4) = 3450 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 75 𝑘𝑔 = (12.32 𝑚𝐿)(75 𝑘𝑔)(4) = 3696 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 80 𝑘𝑔 = (12.32𝑚𝐿)(80 𝑘𝑔)(4) = 3942 𝑚𝐿

For pH = 7.05

𝑉𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 7.05 𝑎𝑛𝑑 7.45 𝑝𝐻 = 15.22 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 50 𝑘𝑔 = (15.22𝑚𝐿)(50𝑘𝑔)(4) = 3044 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 55 𝑘𝑔 = (15.22 𝑚𝐿)(55 𝑘𝑔)(4) = 3348 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 60 𝑘𝑔 = (15.22 𝑚𝐿)(60 𝑘𝑔)(4) = 3653 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 65 𝑘𝑔 = (15.22 𝑚𝐿)(65 𝑘𝑔)(4) = 3957 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 70 𝑘𝑔 = (15.22 𝑚𝐿)(70 𝑘𝑔)(4) = 4262 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 75 𝑘𝑔 = (15.22 𝑚𝐿)(75 𝑘𝑔)(4) = 4566 𝑚𝐿

𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑡 80 𝑘𝑔 = (15.22 𝑚𝐿)(80 𝑘𝑔)(4) = 4870 𝑚𝐿