Osmotic Regulation 1

Osmotic Regulation: Effect of Fasting and Ingestion of Water, Coke, or Gatorade on

Urine Flow Rate and Specific Gravity

Melanie Grajek

Biology 213-07

San Francisco State University

 Osmotic Regulation 2

Introduction:

The function of the kidneys is to maintain water balance, plasma volume, plasma

osmolarity, and acid base balance in the body through the excretion of substances from

filtrate as they turn into urine. The nephron is the functional unit of the kidneys. It is

made up of the afferent arteriole, efferent arteriole, the glomerulus, the Bowman’s

capsule, the proximal convoluted tubule, the Loop of Henle, the distal convoluted tubule,

and the collecting duct. Peritubular capillaries run along the nephron intertwining with

the different sections of the nephron. This is where glomerular filtration, tubular

reabsorption, and tubular secretion occur.

Glomerular filtration is the passive movement of plasma from the walls of the

glomerulus into the Bowman’s capsule, which is first site of urine production.

Glomerular filtration is driven by hydrostatic pressure pushing the filtrate from the

Bowman’s capsule and into the proximal convoluted tubule. Tubular reabsorption occurs

in the proximal convoluted tubule where 99ml/min of water and solutes are reabsorbed

from the tubules back into the peritubular capillaries, while 100% of glucose is

reabsorbed at the proximal convoluted tubule via the sodium-glucose secondary active

transport. There is no hormone regulation at the proximal convoluted tubule, but rather in

the distal convoluted tubule. Aldosterone is a steroid hormone that acts primarily in the

renal collecting ducts to stimulate sodium reabsorption when baroreceptors detect low

blood pressure (Muto, 1995). Aldosterone is produced by the renin-angiotensin-

aldosterone-system (RAAS). In the kidney, juxtaglomerular cells, located at the afferent

arteriole, detect low blood pressure and low blood volume. These cells produce and

secrete the enzyme renin when the body’s blood pressure drops. The liver secretes
 Osmotic Regulation 3

angiotensinogen which is catalyzed by renin when making angiotensin I. The lungs

secrete angiotensin converting enzyme (ACE) which converts angiotensin I to

angiotensin II. This stimulates the release of ADH and triggers the adrenal cortex to

release aldosterone. Aldosterone is secreted from the adrenal cortex in response to low

blood pressure, low blood volume, and low plasma osmolarity. Once secreted, it raises

blood pressure, raises blood volume and raises plasma osmolarity. This will result in a

decrease urine flow rate because the body is absorbing more water instead of releasing,

which in turn also raises urine corrected specific gravity because the body is holding onto

water and thus making it more concentrated.

Anti-diuretic hormone (ADH) stimulates water reabsorption by stimulating

insertion of aquaporin channels into the membranes of the kidney tubules. These

channels transport the water back to the blood, leading to a decrease in plasma

osmolarity. Osmolarity is regulated by osmolreceptors, which stimulate the neurons that

produce ADH when the body’s plasma osmolarity is too high (Bowen). ADH is

synthesized in the supra-optic nuclei and in the para-ventricular nuclei in the

hypothalamus. The release of ADH results in a raise in blood pressure, a raise in blood

volume and a decrease in plasma osmolarity. The results trigger a decrease in urine flow

rate and an increase in urine corrected specific gravity.

Atrial natriuretic peptide (ANP) is a hormone secreted and produced by cardiac

atrial cells in response to an increase in blood volume. Its purpose is to inhibit sodium

and water reabsorption and gets rid of sodium in the body by increasing water excretion

(Zeidel, 1990). ANP’s target sides are the distal convoluted tubule, the collecting duct,

and all the places where ADH and aldosterone are secreted and produced. The results of
 Osmotic Regulation 4

the secretion of ANP are a decrease in blood volume, and increase in glomerular filtration

rate and an increase in urine production. An increase in glomerular filtration occurs

because ANP causes vasodilation of the afferent arteriole. This allows more blood to

enter the glomerulus and allows plasma to filter at a faster rate. In turn, the corrected

specific gravity is decrease because of the urine is diluted and an increase in urine flow

rate.

Tubular section is the secretion of selective substances from the peritubular

capillaries into the proximal and/or distal convoluted tubules to be excreted into the urine

(Sherwood, 2015). If the blood is experiencing blood acidosis, the kidneys will increase

the secretion of hydrogen protons from the peritubular capillaries to get the blood pH

back to normal range. Blood acidosis can be caused by a high increase of hydrogen ions

produced by vigorous exercise, which increases the amount of lactic acid,

hypoventilation, severe diarrhea, and a high fat or protein diet.

The purpose of this experiment was to study the osmotic regulation of the kidneys

in maintaining fluid balance in the body and analyze and compare the changes in urine

flow rate and specific gravity during fasting and following an ingestion of water, Coke or

Gatorade. The purpose was also to test a urine sample for blood, ketones, glucose,

protein, and pH levels using the Labstix strip (Manuguid, 2018). Water, Coke, and

Gatorade were tested because they all have various tonicities compared to plasma. Water

is hypotonic to plasma, Coke is hypertonic to plasma, and Gatorade is isotonic to plasma.

Having three different drinks, each one having a different tonicity, would make it easier

to compare their effect on the plasma because they are so distinctive from each other. My

hypothesis for average urine flow rate was that water would have the highest urine flow
 Osmotic Regulation 5

rate, second Coke, then Gatorade and lastly non-drinking group would produce the lowest

urine flow rate. For average corrected urine specific gravity, I hypothesized non-drinking

would produce the highest urine specific gravity, second water, next Coke, and lastly

Gatorade would produce the lowest urine specific gravity. For the Labstix strip test, I

hypothesized that there would be no trace of blood, ketones, protein, or glucose in the

urine sample and a pH within the normal range of 4.5-8.

Methods:

On the night before the experiment, heavy exercise should be limited as well as

salt, sugar, and caffeine and alcohol consumption. These can all alter the data and have an

effect on the overall class data. Five hours before the lab began, there should have been

no intake of food or drink of any kind and a refrain from any type of vigorous heavy

exercise. If there was any discomfort, there could have been an intake of up to 8oz of

water during the first two hours of fasting. One hour before the lab, the bladder must have

been emptied with the time recorded. Once in lab and the drinking/non-drinking group is

assigned, the amount of fluid for the drinking groups was be calculated. The amount of

fluid intake depended on body weight, so it was specific for each person.

Volume of fluid intake (mL) = [Body weight (lbs) x 7 mL/lbs] x 0.08

At the time of the experiment, T=0, the bladder was emptied into the cups provided and

amount of urine voided and the time was recorded. As soon as possible, the urine

temperature was taken so it was most accurate by not giving the urine time to cool down,

along with duration of time since last void, measured specific gravity and the calculated

corrected specific gravity. Corrected specific gravity is calculated by taking the

 Osmotic Regulation 6

temperature of the urine sample and adding 0.001 for every 3 degrees above 15 degrees

of the urine temperature taken from the urinometer. Urine flow rate was also calculated

after each void.

Flow rate (ml/min) = Volume voided (mL)/Duration of time since last void (min)

These steps were repeated four more times. However, during T=0, a Labstix strip was

used to test if there was any presence of blood, ketones, glucose, proteins in the urine and

the pH of the urine (Manuguid, 2018).

Results:

Average Urine Flow Rate Under Different

Conditions
7
Urine Flow Rate (mL/min)

6
5
4
3
2
1
0
T=0 T=30 T=60 T=90 T=120
Time (minutes)

Non-Drinker Water Coke Gatorate

Fig. 1.1 At T=0 all of the groups had about the same urine flow rate. At T=30, drinking
groups, water, Coke, and Gatorade, all had a significant increase in urine low rate until
T=90. At T=90 minutes, all drinking groups, reached their peak urine flow rate and then
experienced a decrease at T=120. The non-drinking group had a continuous slight decline
at each time interval.
 Osmotic Regulation 7

Average Corrected Urine Specific Gravity

Under Different Conditions
Corrected Urine Specific Gravity

1.035
1.030
1.025
1.020
1.015
1.010
1.005
1.000
T=0 T=30 T=60 T=90 T=120
Time (minutes)

Non-drinker Water Coke Gatorade

Fig. 1.2 Drinking groups, water, Coke and Gatorade, and the non-drinking group
experienced about the same average corrected urine specific gravity until T=30. After
T=30, the drinking groups experienced a drop in corrected urine specific gravity. The
non-drinking group, Coke, and Gatorade experienced a very slight increase between
T=90 and T=120 while the water group experienced a fairly big increase during the same
time.

Urine Labstick Analysis

Presence of Presence of Presence of Presence of Urine pH

Blood Ketones Glucose Protein

Negative Negative Negative Negative 6.0

Fig. 1.3 Presence of blood, ketones, glucose, and protein were all negative. Urine pH was
at 6.0.

Discussion:

Non-drinker:

All students came into class dehydrated; therefore the plasma osmolarity would

be high from high solute concentration. At T=0, the average urine flow rate was at

0.94mL/min. ADH was secreted to reabsorb water, which lowered plasma osmolarity.
 Osmotic Regulation 8

ADH only has a half-life of 16-18 minutes. Since it only lives for a short period of time,

by the time 30 minutes passes, there is a little bit of urine produced instead of it all being

continuously reabsorbed. At T=30 average UFR was 0.85mL/min and at T=60, average

UFR was 0.67mL/min. The lowest average UFR was at T=120 with 0.44mL/min. Low

amounts of hormone ANP was being released from T=30 through T=120 to raise blood

pressure and blood. The average CSG stayed about the same through out the entire

experiment. At T=0 CSG was at 1.026 and raised to 1.029 from T=30 to T=90. At

T=120, CSG increased to 1.030.

Water:

The water group had low plasma osmolarity, high blood pressure and high blood

volume. At T=0 the average UFR was 0.84mL/min so low amounts of ADH were

secreted at T=30 to increase plasma osmolarity and lower blood. This increased average

UFR from 0.84mL/min to 1.45mL/min. The low blood volume and high plasma

osmolarity resulted in a decrease in blood pressure and an increase of UFR. The average

UFR at T=60 was 4.16mL/min. At T=90, high amounts of ADH were secreted to lower

plasma osmolarity resulting in a 5.69 mL/min average UFR which lowered at T=120 to

3.84mL/min. The CSG decreased for the most part through out the experiment. At T=0

average CSG was at 1.030 and decreased to 1.024 at T=30, and then decreased to 1.015 at

T=60. At T=90, CSG continued to decrease to 1.014 and finally raised to 1.020 at T=120.

Coke:

At T=0, the Coke group had very high plasma osmolarity, high blood volume and

high blood pressure. The average UFR was 0.86 mL/min. At T=30, very high amounts of

ADH were secreted to lower plasma osmolarity and UFR resulting in a 0.86 mL/min. A
 Osmotic Regulation 9

high amount of insulin was secreted to tell muscle cells to uptake glucose. This kept the

blood volume and blood pressure high. ANP was secreted to decrease the high blood

pressure and low amounts of ADH were secreted to raise plasma osmolarity and lower

blood volume. At T=60, the average UFR was increased to 3.4 mL/min and at T=90,

UFR raised along to 5 mL/min with blood pressure because the caffeine in coke finally

started to take effect. At T=120, the UFR lowered backed down to 3.88 mL/min. The

average CSG for Coke continuously decreased from T=0 to T=90 from 1.028 to 1.027

and then 1.015 to 1.014. At T=120, CSG slightly increased to 1.016.

Gatorade:

The Gatorade group also had high plasma osmolarity, high blood volume and

high blood pressure at T=0 with a UFR of 0.76 mL/min. ADH is secreted to lower

plasma osmolarity while sodium from the Gatorade continued to increase blood pressure

and blood volume. At T=30, the UFR was raised to 1.04 mL/min. Low amounts of ADH

were being secreted so plasma osmolarity increased, while blood pressure and blood

volume decreased which dramatically increased URF to 4.46 mL/min at T=60. ANP

secretion increased glomerular filtration rate and UFR as a result of high water and

sodium levels. The UFR continued to increase until T=90 where its peak was reached at

5.98 mL/min and then back down to 3.88 mL/min at T=120. Average CSG was at 1.028

at T=0 and at T=30 was at 1.029. It then decreased at T=60 to 1.017 and then again at

T=90 to 1.013. At T=120, CSG slightly increased to 1.014.

My hypothesis’s was rejected. I hypothesized that the non-drinking group would

have the lowest urine flow rate, which was correct. However, I hypothesized that water

would have the highest urine flow rate, but Gatorade had the highest UFR, then water,
 Osmotic Regulation 10

and coke as the second lowest. For corrected specific gravity my hypothesis was also

rejected. Non-drinkers had the highest CSG, but following the non-drinking group, the

second highest was Coke, not water, then Gatorade, not Coke, and water as the lowest.

The purpose of avoiding food and drink before the lab began was to have all of

the subjects at about the same level of hydration. Since the entire class, in theory, was

dehydrated, there would be a more dramatic result when big amount of liquids were

consumed. Each hormone reacts to different stimulus in the body so we would be able to

see how each hormone reacts under each condition. For example, aldosterone and anti

diuretic hormone secretion occurs when blood volume is low while atrial natriuretic

peptide hormone secretion occurs when blood volume is high. The hormones work to

continuously help the body reach homeostasis.

My hypothesis for the urine Labstix analysis was supported: in that there would

be no presence of blood, ketones, glucose or protein in my urine. All of the results came

back negative and I had a pH of 6.0, which is in the normal range. There was no presence

of blood, protein, ketones and glucose because it was all filtered out or reabsorbed during

glomerular filtration. A physiological factor that would reject my hypothesis about

proteins in my blood would be if I had proteinuria, a renal dysfunction, or kidney disease.

For glucose, trace amounts could be found in the urine if I was pregnant, was under great

emotional stress or have glycosuria as a result from hyperglycemia associated with

diabetes mellitus. A trace of ketones would show that the body is using fat as the main

source of energy instead of carbohydrates. It could also mean that the person has diabetes

mellitus, is starving, vomiting, fasting, or eating an all protein diet. Blood in the urine

would indicate that there is a leak in the glomerular membrane, not allowing blood to be
 Osmotic Regulation 11

filtered out, an allergic reaction or they are experiencing hematuria, a symptom of renal

diseases and disorders of genitourinary system. Traces of blood in the urine could also be

from menstruation.

According to the study done by Grandjean (2000), those who drank water

compared to those who drank a carbonated beverage and water, the results were about the

same as the ones done in class. The result from the study was done pre and post-

treatment morning voids, and the results had no significant changes. For water, the urine

specific gravity read 1.019 ± 0.006 about the same results the class received, and for

carbonated beverage it was also 1.019 ± 0.005. There was more of a difference in the

class data but that is probably because the Coke group did not also ingest an equal

amount of water.

Conclusion:

This lab showed how the kidneys maintain fluid balance in the body by

comparing the changes in urine flow rate and specific gravity during fasting and

following an ingestion of water, Coke or Gatorade. The Gatorade group produced the

highest urine flow rate and the non-drinker group produced the highest corrected specific

gravity. In the future, it would be interesting to have a control group who did not fast or

change anything about this diet to see under normal conditions how the Gatorade, water,

and Coke, effects their UFR and CSG.

 Osmotic Regulation 12

References

Bowen, R. Physiologic Effects of Antidiuretic Hormone. Retrieved April 18, 2018.

http://www.vivo.colostate.edu/hbooks/pathphys/endocrine/hypopit/adh.html

Grandjean, Ann. (2000). The Effect of Caffeinated, Non-Caffeinated, Caloric and Non-

Caloric Beverages on Hydration. Vol. 19, NO. 5.

Manuguid, Christine (2018). Experiments in Principles of Human Physiology.

Muto, S. (1995). Action of Aldosterone on Renal Collecting Tubule Cells. Retrieved

April 13, 2018. https://www.ncbi.nlm.nih.gov/pubmed/7743155

Sherwood, L (2015). Human Physiology: From Cells to System (9th edition). Cengage

Learning.

Zeidel, M (1990). Renal Actions of Atrial Natriuretic Peptide: Regulation of Collecting

Duct Sodium and Water Transport. Vol. 52.

https://www.annualreviews.org/doi/abs/10.1146/annurev.ph.52.030190.003531