Comparison of the effect on blood glucose and

serum antioxidants between pomegranate juice and

beet juice on young adults enrolled in NUTR 302L

Running Head: Pomegranate vs. beet juice on blood glucose and serum

antioxidants

Author: Karina Jaime Carbajal, School of Exercise and Nutritional Sciences, San Diego State University,

San Diego, CA 92182

Corresponding author: Dina Metti, MS; Department of Exercise and Nutritional Sciences; San Diego

State University; 5500 Campanile Drive; San Diego, CA 92182; dmetti@sdsu.edu

Abstract:

NUTR 302 students were participants in a research study conducted within the PSFA laboratory

in San Diego State University. This study was unique in that it was conducted on two separate

days with a week washout period, it only used pomegranate and beet juices, and it tested for

blood glucose and serum antioxidants simultaneously. It aimed to specifically compare the

differences that beet and pomegranate juices had on the blood glucose and serum antioxidant

levels throughout the two hours after consumption in the participants. The 21 participants (N=21)

were required to fast for a minimum of five hours prior to the study, drink the provided juice,

check their blood glucose at baseline, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90

minutes, and 120 minutes, draw blood for later antioxidant measurements at baseline, 60

minutes, 90 minutes, and 120 minutes, have a week washout period, and repeat with the second

juice. The 6 oz. of water with the tablespoon of beet root powder significantly provided more

antioxidants than the 5.7 oz. of pomegranate juice (p <0.05). Both juices affected the blood

glucose levels in similar ways providing no significance (p<0.05). In conclusion, the beet root

powder juice provided more antioxidants than the pomegranate juice while acting similarly on

the blood glucose of the participants.

Key words: beet, pomegranate, juice, glucose, antioxidants

Introduction:

In 2015, three of the ten leading causes of death worldwide had to do with cardiovascular

disease (CVD) and diabetes1. CVD is an umbrella term for the ischemic heart disease (IHD),

stroke, high blood pressure, cardiac arrest, and more. Stroke and IHD were the leading two

causes, and diabetes was in the sixth ranked cause1. For diabetes, cardiovascular complications

increase the diabetes-related morbidity2. All of these diseases that contributed to so many deaths

worldwide are connected and affect a lot of the same people, meaning that some of the causes

and roots to these diseases are connected as well. Specifically, they all can be stemmed from the

issues of uncontrolled blood glucose and oxidative stress.

What happens in IHD and stroke patients is that low density lipoproteins (LDL) get

deposited into arteries, the LDL gets oxidized by free radicals, the smooth muscle surrounding

arteries grows, artery walls thicken with this plaque, and blood flow gets restricted. With a

restricted blood flow, the heart struggles to pump blood throughout the body, which can cause

the heart to not receive enough of the blood it needs to pump again. Or, the plaque builds up and

then either some of the plaque detaches and travels through the arteries until reaching the smaller

ones which it then clots, or by restricting blood flow at the site of the LDL oxidation. The issue

here overall is the oxidation of LDL which antioxidants can help prevent.

Antioxidants work by having electrons they can freely donate to the free radicals. All the

free radicals want is the electrons; if antioxidants are present to donate the electrons, everything

stays neutral and healthy. However, if antioxidants are not present, then the free radicals take the

electrons they want from LDL which makes it oxidized and increase the risk for the

beforementioned IHD and stroke. This was shown to be true in a study that measured not only
the serum antioxidants in their participants, but also measured the oxidized LDL levels3. They

found that there was indeed the correlation of more antioxidants leading to less oxidized LDL3.

People with type 2 diabetes need to worry about their blood glucose levels. For them, it’s

important to be aware of how juices affect blood glucose levels so that they could pick the one

that would make the lesser spike. This is due to the fact that too much glucose in their arteries,

that isn’t absorbed because of their insulin problems, can cause significant damage to their

arteries. This uncontrolled blood glucose level also increases the risk for CVD related mortality

significantly2. Fiber, however, helps lower this issue.

Fiber works by forming a gel in the stomach and making the gastric emptying rate slow

down. This decrease in speed results in the blood glucose being released into the blood slower

and lessening the severity of high blood glucose levels mentioned above. This relationship was

looked into in a study that used rice bran to try to help stabilize blood glucose4. They found that

participants that were given the fiber-rich rice bran had better blood glucose levels4.

Pomegranates and beetroots are both foods known for having antioxidants and fiber.

Pomegranates have polyphenols that are known to suppress inflammation and help with cancer5.

While this does not show their specific help with LDL, it shows that they are full of antioxidants

with benefits in reducing general oxidative stress. Beet root is also known to have both

polyphenols and flavonoids which also help combat oxidative stress, and it is highly bioavailable

according to a study done by testing a variety of different beetroot juices6. Both fruits are also

high in fiber, yet juicing could make this different; that’s how this study can help add on to the

already existing research.

 Our study is unique and different from these two because we tested for blood glucose and

serum antioxidants simultaneously in attempt to find which juice (between the pomegranate and

beetroot juices) would both not cause a spike in blood glucose while still providing antioxidants.

The serving sizes for the juices were: 5.7oz of pomegranate juice and 6oz of water mixed with

one tablespoon of beetroot powder. Since both the pomegranate and beet juice had high

concentrations of polyphenols, but the beetroot juice was the only one that still contained fiber

and had less sugar, it was hypothesized that both the beet and pomegranate juices would have

affected serum antioxidants in the same way, but that beet juice would have caused a slower and

lesser increase of blood glucose.

Methods:

Study Design

The study design for this research study was a quasi-experimental crossover with a

seven-day washout period. There was no random assignment of juices to participants; they all

drank the same juice on the same day and participated again a week later with the same second

juice.

Participants

The participants were all volunteers that were enrolled in San Diego State University’s

Nutrition 302L. There was a total of 21 participants (N=21) with 5 males and 16 females.

Participants could not participate if they had not met the fasting requirements, had any allergies

to pomegranates of beet root juices, could not handle the blood draw requirements, or were

pregnant at the time of the study.

Procedure

Before beginning the experiment, the participants were told exactly what would happen

on each day of the studies. They were told to fast for the five hours prior to their class period

where the experiment would begin. Before the class started, students not participating in the

study were asked to help prepare the juice that would be tested on that day. When the

participants came in, they first checked their baseline blood glucose levels using a Medtronic

glucometer and, separately, collected approximately 400uL of their blood in small heparinized

vials. They then drank the juice that would be tested for that day. After 15, 30, and 45 minutes,

they checked their blood glucose levels with the glucometer. At the 60, 90, and 120 minute

markers, they checked their blood glucose again and collected an additional 400uL of their blood
each of those times. Every blood glucose draw and blood collection was done with a finger prick

flowed by pumping the finger to get enough blood for either or each test. The heparinized vials

with the 400uL of blood were each centrifuged at 4,000 RPM for 10 minutes by the students that

did not participate in the study. Then, the plasma was collected and stored in Eppendorf tubes at

-70 °C for no more than 2 weeks. This process was then repeated a week later with the second

juice.

At the end of the study, the antioxidants in the serum were tested for. Again, this was for

the four blood collections done each day of the study (at baseline, 60, 90, and 120 minutes). The

Trolox Equivalent Antioxidant Capacity (Cayman Chemical, Ann Arbor, Michigan) assay kit

was used for this. Metmyoglobin was added to the 2,2’-azino-bis (3-ethylbenzthiazoline-6-

sulfonic acid) (ABTS). Since the metmyoglobin is an oxidation chemical, it would change the

ABTS into ABTS+. The antioxidants from the juices, however, would prevent the ABTS

oxidation at different levels depending on its concentration and ability. This is why the serum

and metmyoglobin mixture was read at the 405nm absorbance. The lighter the serum was, the

better the antioxidants worked at stopping the ABTS to ABTS+ reaction.

Statistical Analysis

The data were analyzed using repeated measures in analysis of variance (ANOVA). Since

it was analyzed after all the data were collected, it was considered to be analyzed in the

Bonferroni post-hoc manner. Both the glucose and antioxidant levels were intervened in during

the study via the juices and analyzed for. The alpha value was set at p<0.05 so that any data had

to have a p-value of (p<0.05) to be considered significant. After finding significance (p<0.05) for

the antioxidants, a pairwise comparison test was also done to find where exactly the significance

was.
Results:

The participants used in this study were an average of 25 years old give or take 7.42

years. The average weight was 142.2±27.2 pounds. The average height was 66±22.04 inches.

Finally, the average body mass index (BMI) was 22.9±3.32 kg/m2.

In regard to the blood glucose, the beet and pomegranate juices at each of the seven time

markers (baseline, 15, 30, 45, 60, 90, 120 minutes) showed no significant (p<0.05) difference.

The averages for the baseline, 15, 30, 45, 60, 90, and 120-minute blood glucoses after the

pomegranate juice consumption were 100.0952±14.876mg/dL, 122.524±mg/dL, 139.429±20.388

mg/dL, 121.429±22.218 mg/dL, 106.857±24.502 mg/dL, 94.762± mg/dL, and 95.619±10.703

mg/dL respectively (Figure1). The averages for the baseline, 15, 30, 45, 60, 90, and 120-minute

blood glucoses after the beetroot juice consumption were 102.053±13.705 mg/dL,

128.684±22.480 mg/dL, 138.579±27.187 mg/dL, 128.263±17.502 mg/dL, 112.737±16.041

mg/dL, 109.000±21.520 mg/dL, and 110.842±14.901mg/dL respectively (Figure 1). A bar graph

showing the data for both the juices’ blood glucose effect can be seen in Figure 2 of the

appendices.

In regard to the antioxidant level, the beet and pomegranate juices did show significance

(p<0.05) difference between both the baseline and 90-minute mark and between the baseline and

120-minute mark. The averages for the baseline, 60, 90, and 120-minute serum antioxidants after

the pomegranate juice consumption were 2.142±1.679mM, 2.496±2.162mM, 2.560±2.068mM,

and 1.979±0.848mM respectively (Figure 3). The averages for the baseline, 60, 90, and 120-

minute serum antioxidants after the beetroot juice consumption were 1.684±0.609mM,

1.636±0.648mM, 3.374±2.026mM, and 3.856±2.299mM respectively (Figure 3). A bar graph

showing the data for both the juices’ serum antioxidant effect can be seen in Figure 4 of the

appendices.
Discussion:

Since the two juices did not act significant (p<0.05) on blood glucose, and the beetroot

juice was the only one that contained fiber and less sugar, the reasons for the blood glucose

similarity must have been of how the insulin worked in the participants. Insulin is typically

secreted after food is ingested and acts on one’s muscles, liver, and fat cells. It allows these

different cells to take in the glucose so that it does not continuously circulate in the blood. In the

muscles, liver, and fat, the glucose can then be used for energy or stored. When these juices were

ingested by the participants, the insulin must have been released properly and worked throughout

the two hours. Generally, the blood glucose levels rose during the first half hour and then

decreased for everyone and for the both juices by the 45-minute marker. This shows that the

insulin was actively working between the 30 to 45 minute markers. Since the blood glucose

continued to drop until the 90-minute marker, that is when the insulin had finished working.

From there, the blood glucose increased slightly again in the body’s natural response to keep the

blood glucose at a steady level. In sum, the fiber and amount of sugar in these juices did not

change the blood glucose too differently. Both juices acted similarly. And this can possibly be

due to the insulin working reliably in all the participants.

The two juices, fortunately, did act statistically (p<0.05) different when it came to how

they affected serum antioxidant levels. Specifically, significance (p<0.05) was found when

comparing the baseline to the 90 and 120 minute markers. From the pomegranate juice, the

values show that the serum antioxidant levels did increase from baseline to the hour mark, but

then decreased back to near the baseline by the end of the second hour. This could mean that the

amount of antioxidants in the serum increased after consumption, but were then either used or

destroyed. For the beetroot juice, on the other hand, after the hour mark, the serum antioxidant
levels continually increased a lot. This could mean that they were more slowly absorbed, that

they were stronger forms of antioxidants, that they were less needed and used, or some

combination of the above. All in all, the beetroot juice increased serum antioxidants much better

than the pomegranate juice.

For those with type 2 diabetes who must worry about their blood glucose levels, it would

be important for them to know that the two juices acted similarly on subjects with normal-

working insulin. With neither juice affecting the blood glucose statistically different (p<0.05),

they should be able to drink either and not have to worry too much about one giving them a

higher or longer blood glucose peak. If they keep their diabetes in control in whatever way

recommended to them, no juice should cause them greater difficulties than the other in

preventing the blood glucose from damaging their arteries. This conclusion is limited, however,

since the data were collected using only participants without diabetes. Since fiber did affect

participants differently in the rice-bran study4, more research needs to be done.

For all peoples, including those with IHD or who are at higher risk for strokes, the

beetroot juice that provided more antioxidants is the better choice of juice. Since the antioxidants

increased more, there would be more antioxidants to help decrease the amount of LDL that gets

oxidized and could become fatal. Especially since the blood glucose does not need to be

considered in the juice decision, based off of antioxidants, the beetroot juice provided the most

protection against arthrosclerosis, heart diseases, and strokes. Since these are among leading

causes of death worldwide1, this finding could be greatly beneficial. And since diabetes was on

the leading causes as well1 and cardiovascular contributions make it more fatal2, this finding is

equally beneficial for them. This study was limited, however, because it only checked the
antioxidant levels and did not specifically check the oxidized LDL levels like the pistachio study

did3.

In future research, this study could be expanded upon by including a diabetic group, by

also checking for oxidized LDL levels, by checking which specific antioxidants were present, by

checking the antioxidant levels for longer than the two hours, by using beetroot juice from beets

as opposed to from a powder, and by having an overall larger sample size.

In conclusion, this study found that beetroot juice has a significantly (p<0.05) different

amount of antioxidants than pomegranate juice while not have significant (p<0.05) changes to

blood glucose. The beetroot juice had the higher number of antioxidants, and, thus, is the better

choice of juice when selecting one to help with lessening oxidative stress. Since the study was

unique in simultaneously checking the effect of two juices on blood glucose and serum

antioxidants, it allowed for a new niche of studies to be done and expanded upon. This research

is critical in helping lower the morbidity to some of the leading causes of death worldwide.
References

1. Top 10 Causes of Death. WHO Web site.

http://www.who.int/mediacentre/factsheets/fs310/en/. Published January 2017. Accessed

May 1, 2017.

2. Vazquez-Benitez G, Desai J, Xu S, Goodrich G, Schroeder E, et al. Preventable major

cardiovascular events associated with uncontrolled glucose, blood pressure, and lipids and

active smoking in adults with diabetes with and without cardiovascular disease: A

contemporary analysis. Diabetes Care. 2015;38(5):905.

3. Kay C, Gebauer S, West S, Kris-Etherton P. Pistachios increase serum antioxidants and

lower serum oxidized-ldl in hypercholesterolemic adults. The Journal of Nutrition.

2010;140(6):1093-1098.

4. Wu T, Chen W. The effect of rice-bran dietary fiber on postprandial blood glucose for type ii

diabetic patients. Diabetes Research and Clinical Practice. 2016;(suppl)120:S103.

5. Gözlekçi S, Gozlekci S, Saraçoglu O, Onursal E, Özgen M. Total phenolic distribution of

juice, peel, and seed extracts of four pomegranate cultivars. Pharmacognosy Magazine.

2011;7(26):161.

6. Guldiken B, Toydemir G, Memis K, Okur S, Boyacioglu D, et al. Home-processed red

beetroot (beta vulgaris l.) products: changes in antioxidant properties and bioaccessibility.

International Journal of Molecular Sciences. 2016;17(6):858.

Appendix:

Figure 1:

Blood glucose levels at specified time intervals after pomegranate and beetroot juice

consumption in the NUTR 302L participants.

0 min 15 min 30 min 45 min 60 min 90 min 120 min

Avg. 100.0952 122.5238 139.4286 121.4286 106.8571 94.7619 95.6190
(mg/dL) for
pomegranate
Std Dev 14.8758 14.9219 20.3877 22.2184 24.5016 14.8220 10.7027
(mg/dL) for
pomegranate
Avg 102.0526 128.6842 138.5789 128.2632 112.7368 109.000 110.8421
(mg/dL) for
beetroot
Std Dev 13.7051 22.4798 27.1872 17.5019 16.0411 21.5200 14.9006
(mg/dL) for
beetroot
Figure 2:

Blood glucose concentrations shown at specified time intervals for the pomegranate and beetroot

juices with standard deviations shown for the participants in the 302L study.

180

160

140

120
Concentration

100

80

60

40

20

0
0-min 15-min 30-min 45-min 60-min 90-min 120-min
Time

Pomegranate Beet Root Powder

Figure 3:

Serum antioxidant levels at specified time intervals after pomegranate and beetroot juice

consumption in the NUTR 302L participants.

0 min 60 min 90 min 120 min

Average (mM) 2.1416 2.4956 2.5601 1.9785
for pomegranate
Std Dev (mM) 1.6789 2.1629 2.0685 0.8484
for pomegranate
Average (mM) 1.6839 1.6359 3.3745 3.8559
for beetroot
Std Dev (mM) 0.6091 0.6481 2.0260 2.2986
for beetroot
Figure 4:

Serum antioxidant concentrations shown at specified time intervals for the pomegranate and

beetroot juices with standard deviations shown for the participants in the 302L study with

significance (*, p<0.05).

5 *
Concentration

0
0-Min 60-Min 90-Min 120-Min
Time

Pomegranate Beetroot