Glucose and Insulin Responses to the Consumption of Dried Goji Berries and Almonds

Running head: Glucose and Insulin Response to Phytochemicals

Author:
Macy Wright
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

Glucose and Insulin Response to Phytochemical Wright 1

Abstract
Berries and tree nuts have been widely researched in terms of their many health benefits,
including the prevention of chronic illnesses such as cardiovascular disease and type 2 diabetes.
Berries and tree nuts include bioactive compounds, such as anthocyanin, that have been shown to
benefit the health of those who consume them. There have been research done on both tree nuts
and berries separately, but there has not been conclusive research done on the effects of
consuming tree nuts and berries together. The purpose of this study was to find the effects of
both dried goji berries and almonds together on blood glucose and insulin levels in the body. This
was done by carrying out a randomized controlled crossover study with the SDSU students
enrolled in Nutrition 302L. Participants were to fast and consume different foods, one piece of
white bread, 28 grams of dried goji berries, and 28grams of dried goji berries and 30 grams of
almonds. Blood was taken and results were found and compared. It was found that the
consumption of dried goji berries and almonds together had the best results in terms of
improvements of glucose tolerance and insulin resistance. ANOVA tests were done, then
Bonferroni follow up testing was done in order to find significance of the findings. There was
significance found from both the ANOVA tests and the Bonferroni tests. In conclusion, the
hypothesis that the dried goji berries and almonds consumed together would result in better
blood glucose and insulin levels than white bread would, was supported from the findings and
data of this study.
Glucose and Insulin Response to Phytochemical Wright 2

Introduction

Many plant foods are known to contain certain phytochemicals, which have been found

to aid in the prevention of chronic diseases such as cardio vascular disease, cancer, diabetes

mellitus, and several others. While these phytochemical can be found in many foods, they are

especially plentiful in fruits and vegetables.1 Two of the main causes of death in the United

States are actually cancer and cardiovascular disease, which proves the importance of

incorporating an adequate amount and variety of fruits and vegetables into the diet. Many

chronic diseases are caused by an overproduction of oxidants in the body, in which the

antioxidant properties of phytochemicals found in plant sources can help reduce.2 It is more

effective to try and prevent chronic illnesses than it is to treat them, and simply consuming fruits

and vegetables containing phytochemicals could prevent some of these illnesses and deaths.3

It has been shown that certain berries have compounds that have antioxidant properties

when consumed. Blueberries, for example, contain different sources of polyphenols such as

anthocyanin. Anthocyanins are the pigments that are also found in many other deep blue, red,

and purple fruits and vegetables, including goji berries.4 Anthocyanin has been shown to

decrease insulin resistance and improve glucose tolerance when consumed.5 The addition of

fruits and vegetables containing anthocyanin to the diet, such as blueberries or goji berries, could

greatly benefit those with type 2 diabetes. Type 2 diabetes is caused by high levels of insulin

resistance and glucose levels in the body. Other benefits of the phytochemicals found in berries

include anti-aging properties, anti-inflammatory properties, reduced risk of other chronic

diseases, improved gastrointestinal health, and weight maintenance.6

Glucose and Insulin Response to Phytochemical Wright 3

Another food that contains many phytochemicals that have been shown to aid in several

health benefits and prevention of diseases is tree nuts, such as almonds. Tree nuts contain

phenolic acids, phytosterols, polyphenols, and other phytochemicals that have antioxidant, anti-

inflammatory, and hypocholesterolemic properties in the body when consumed regularly.7 Tree

nuts also contain unsaturated fatty acids in which aid in glycemic control and appetite

suppression. Both of these properties could be beneficial for those with type 2 diabetes, along

with the effect that the fiber and polyphenols in tree nuts have on improving gut microbiota. The

regular incorporation of almonds or other tree nuts into the diet could improve oxidative stress,

blood pressure, glucose homeostasis, and weight maintenance, along with aiding in the

prevention of chronic illnesses such as cardiovascular diseases or diabetes type 2 diabetes.8

There have been many studies done on the health benefits and aid in prevention of

chronic illnesses of phytochemicals in berries and other plant sources, such as tree nuts. Both

berries and tree nuts have antioxidant properties that have been shown to help with the regulation

of insulin resistance and glucose levels in the body. There have not, however, been any studies

done on how the consumption of both berries and tree nuts consumed together effect insulin

resistance and glucose levels, which was the purpose of this study. During the study, participants

were given different groups of food to consume after fasting, with the control being white bread.

Blood samples were taken before and after consumption of the foods in order to test blood

insulin and glucose levels and any changes shown in each. It was hypothesized that the

consumption of dried goji berries along with almonds would stabilize glucose and insulin levels

more than the control, white bread, would.

Glucose and Insulin Response to Phytochemical Wright 4

Methods

Study Design

The study design used was a randomized controlled crossover design in which each

participant was randomly assigned to one of three groups on the first day of the study. The

control group was the white bread. This study design was used so that the information gathered

was accurate and each participant tried each of the three groups. Between each trial, there was a

one day washout period.

Participants

The subjects used for this study were all San Diego State University students that were

enrolled in the Nutrition 302 Lab course during the time of the study. There were twelve

participants used, in which each was given one of three groups on the first day. Certain

exclusions were included in terms of who could participate in the study, which included those

who could not meet the fasting requirements given, those who were allergic to any of the foods

used in the study, or those who were pregnant during the trials.

Procedure

On the first day of the study, each of the twelve participants were given one of three

groups. The three groups were the control, white bread, dried goji berries, and dried goji berries

and almonds. The participants were each given a standard meal around fifteen minutes before the

five hour fast started. The standard meal was a peanut butter and jelly sandwich, consisting of

two pieces of white bread, two tablespoons of plain peanut butter, and two tablespoons of

strawberry jelly.
Glucose and Insulin Response to Phytochemical Wright 5

The three groups included in the study were the white bread, the goji berries, and the goji

berries and almonds. The first group was given one slice of white, Wonder Brand, bread. One

slice of white bread includes 60 calories, 1.5 grams of fat, 4 grams of protein, 24 grams of

carbohydrates, and less than 1 gram of fiber. The second group was given 28 grams of dried goji

berries, Made in Nature brand. This includes 100 calories, 22 grams of carbohydrates, 0 grams of

fat, 3 grams of protein, 22 grams of carbohydrates, and 5 grams of fiber. The third group was

given 28 grams of dried goji berries, Made in Nature brand, and 30 grams of raw almonds, Blue

Diamond brand. This includes a total of 260 calories, 28 grams of carbohydrates, 14 grams of fat,

9 grams of protein, and 8 grams of fiber.

After the five hour fast, the participants each gave a 400uL sample of blood to check

glucose levels. This check was the 0 minute mark. After the blood was taken, each participant

were given five minutes to eat their snack, depending on which group they were assigned to.

After the snack, glucose levels were checked again after 15 minutes, 30 minutes, 45 minutes, 60

minutes, 90 minutes, and 120 minutes. After the snack was eaten, 400uL samples of blood were

agin taken by each participant atfter 30 minutes, 60 minutes, 90 minutes, and 120 minutes in

order to measure insulin levels. After the last blood samples were taken, the first trial was done,

there was a one day washout period, then the participants switched groups in order for them to

try the next food group for the second trial. These tips were repeated for the second trial, then

there was another one day washout period and the groups switched one last time and the third

trial was done.

Glucose and Insulin Response to Phytochemical Wright 6

Biochemical Data

In order to measure glucose, each participant had their fingers pricked and a Medtronic

glucometer and Nova Max test strips were used to measure the glucose. In order to measure

insulin levels, insulin ELISA kits were used by the brand ALPCO. Once the blood samples were

taken, they were stored in heparinized vials and centrifuged at 4000RPM for 10 minutes. After

the 10 minutes, plasma was collected from the blood and stored in eppendorf tubes at -70

degrees Celsius for up to two weeks.

Statistical Analysis

The data found in this study was analyzed using a mixed ANOVA test. For the glucose

ANOVA, there were three treatments tested against seven different time points. For the insulin

ANOVA there were three treatments tested against five different time points. The two

components compared in each of the tests were time and the three treatments. In order to

determine whether there was significance, the p-value needed to be less than 0.05. The software

used was the SPSS SOFTWARE Version 26. Then if there was significance found in the data,

follow up analysis was necessary, using Bonferroni post-hoc analysis.

Results

The characteristics of the participants of the study are shown in figure 1.1 in the appendix

section. There originally were 12 participants, however, only 7 were included in the study due to

some not being able to fulfill the requirements. One male and six females were included in the

study. The mean height of participants was 158.9 cm, with a standard deviation of 12.0. The

mean weight of participants was 63.6 kg with a standard deviation of 6.0. The mean BMI was
Glucose and Insulin Response to Phytochemical Wright 7

24.7 with a standard deviation of 2.4. The mean age of participants was 29.7 with a standard

deviation of 7.7. The 7 participants included in the study were of good health and were able to

meet the requirements given for the study.

The mixed design ANOVA test done on postprandial effects on glucose levels showed

that there was a significant difference for time (p<0.000) and for time and group (p<0.021). Once

this significance was found from the ANOVA test, follow up Bonferroni testing was necessary in

order to decipher what data was significant. The Bonferroni testing results are shown in figures

1.2 and 1.3 in the appendix section. The Bonferroni test showed both the results of blood glucose

levels within the groups, and blood glucose levels between groups. In terms of blood glucose

levels within the groupd, for white bread (WB) it is shown that there was a significant increase in

glucose levels from 0 minutes to 60 minutes. For dried goji berries (GB), there was a significant

increase in glucose levels from 0 minutes to 15 minutes. For dried goji berries and almonds (A),

there was a significant increase from 0 minutes to 30 minutes, 0 minutes to 45 minutes, and 0

minutes to 60 minutes. As for blood glucose levels between groups, there was a significant

increase for white bread (WB) compared to goji berries (GB) at both 45 minutes and 60 minutes.

The mixed design ANOVA test done on postprandial effects on insulin levels also showed

that there was significance for time (p<0.000). Once this significance was found, there was also

Bonferroni testing done to find what data was significant. Both results of blood insulin levels

within groups and between the groups were tested. The Bonferroni test results for blood insulin

levels are both shown in figures 1.4 and 1.5 in the appendix section. In terms of blood insulin

levels within the groups, there was a significant increase shown in insulin levels for goji berries

and almonds (A) from 0 minutes to 60 minutes. In terms of blood insulin levels between the
Glucose and Insulin Response to Phytochemical Wright 8

groups, there was a significant increase shown in goji berries and almonds (A) compared to goji

berries (GB) at 60 minutes.

Discussion

This study was done in order to see the results and effects on blood insulin and glucose

levels, when consuming one slice of white bread, 28 grams of dried goji berries, and 28 grams of

dried goji berries with 30 grams of almonds. It was hypothesized that the consumption of dried

goji berries along with almonds would stabilize glucose and insulin levels more than the control,

white bread, would, which was shown to be true with glucose levels through this study. This is

not a surprising, considering the high glycemic content of white bread compared to both dried

goji berries and almonds. As shown in figure 1.3, white bread (WB) caused the highest levels of

glucose, dried goji berries (GB) caused the lowest blood glucose concentrations, and both dried

goji berries and almonds (A), caused the most stable blood glucose levels. In terms of the blood

insulin levels, shown in figure 1.5, dried goji berries and almonds (A) actually showed the

highest spike. White bread (WB) was the second highest insulin levels, and dried goji berries

(GB) showed the lowest insulin levels. As for people with insulin resistance issues or high

glycemic levels in their bodies, this information could be very helpful.

Berries that have bioactive compounds, such as anthocyanin, have been proven to be

helpful to consume for those with type 2 diabetes, along with those who wish to decrease the risk

of type 2 diabetes. This is due to the positive effects that anthocyanin have upon insulin

resistance and glucose tolerance, which is a bioactive compound found in dried goji berries.5

This is shown through figures 1.3 and 1.5 in the appendix. Goji berries (GB) caused the lowest
Glucose and Insulin Response to Phytochemical Wright 9

spike in glucose levels out of all of the groups. In terms of insulin resistance, serum levels spiked

and then dropped and stayed steady. This could be explained by the action of the anthocyanin in

the berries causing the consumption of them to regulate glucose and insulin levels. Goji berries,

along with other berries that contain the bioactive compound anthocyanin are shown to be good

for those with type 2 diabetes or those that wish to prevent it.

As for almonds, they have also been proven to be good for the prevention and control of

type 2 diabetes. The unsaturated fatty acids and other compounds improve glucose control and

homeostasis.8 Therefore, when goji berries and almond are eaten together, they should, in theory,

also be very good for glucose and insulin control in diabetics. It was shown in figure 1.3 that

dried goji berries and almonds (A) eaten together causes the most steady levels of glucose in the

body out of all of the groups. In terms of insulin levels, this group spiked insulin levels the most.

These results in the data could be explained by the unsaturated fatty acids in almonds improving

glucose control and therefore, regulating and steadying the glucose levels.

In terms of the way that the study was done, there are some concerns as to its accuracy.

The sample size of the study was originally meant to be 12 participants, but due to five

participants not being able to give the 400uL blood sample in order to test glucose and insulin

levels, there were only 7 participants included. This is not a very big sample size, and if there

were more participants, it might be more accurate. There was also a wide discrepancy in gender

of the participants. There were six women included in the study, and only one man. If there

participants were more evenly spread in terms of gender, this could also increase the accuracy of

the study. There was also no monitoring in terms of the participants fasting, therefore, there is no

way to tell if fasting glucose levels were accurate to begin with or not. The part of the study that
Glucose and Insulin Response to Phytochemical Wright 10

was good is that it was a randomized crossover design. This made it so that the participants were

randomly placed into there three groups, but they switched and got to participate in each group.

This improves the accuracy of the results of the study, since levels for each participant were

measured for each group.

There was some data found that was trending toward significance, but did not quite reach

the p-value (p<0.05). If there was more accuracy within the study, this data could have possibly

reached the p-value (p<0.05) and reached significance. From the glucose Bonferroni test between

groups, white bread (WB) and dried goji berries and almonds (A) trended toward significance at

60 minutes. From the Bonferroni test within groups for insulin, white bread (WB) trended toward

significance from 0 minutes to 60 minutes. The Bonferroni test for insulin between groups shows

that white bread (WB) and dried goji berries (GB) trended toward significance at 60 minutes.

The data found in this study can not be used for the general public due to the accuracy

concerns. The sample size was too small to use the data for the general public, along with the

fact that the study was not as monitored in terms of fasting time accuracy. The study could have

been monitored more. It would also be a good idea to include different participants in the study

with varying levels of health. The main reason for this study was to decide the effects of insulin

and glucose levels in the body, which could be beneficial to those with health issues, such ass

type 2 diabetes. With that being said, the result and data could have been more interesting if there

were people with health concerns included in the study.

The hypothesis that the consumption of dried goji berries along with almonds would

stabilize glucose and insulin levels more than the control, white bread, would, was supported

through the data and shown through the figures in the appendix. The unsaturated fatty acids in
Glucose and Insulin Response to Phytochemical Wright 11

unison with the anthocyanin in the dried goji berries may improve and help regulate glycemic

tolerance and insulin resistance in the body, however, more testing needs to be done on the

subject in order to prove these findings.

Glucose and Insulin Response to Phytochemical Wright 12

References

1. Chen L, Vigneault C, Raghavan V, Kubow S. Importance of the phytochemical content of

fruits and vegetables to human health. Stewart Postharvest Review. 2007;3(3):1–5.

2. Zhang Y-J, Gan R-Y, Li S, Zhou Y, Li A-N, Xu D-P, et al. Antioxidant Phytochemicals for
the Prevention and Treatment of Chronic Diseases. Molecules. 2015;20(12):21138–56.

3. Liu RH. Health benefits of fruit and vegetables are from additive and synergistic
combinations of phytochemicals. The American Journal of Clinical Nutrition. 2003;78(3).

4. Khoo HE, Azlan A, Tang ST, Lim SM. Anthocyanidins and anthocyanins: colored pigments
as food, pharmaceutical ingredients, and the potential health benefits. Food &amp; Nutrition
Research. 2017;61(1):1361779.

5. Stull A. Blueberries’ Impact on Insulin Resistance and Glucose Intolerance. Antioxidants.

2016;5(4):44.

6. Kalt W, Cassidy A, Howard LR, Krikorian R, Stull AJ, Tremblay F, et al. Recent Research on
the Health Benefits of Blueberries and Their Anthocyanins. Advances in Nutrition. 2019;

7. Bolling B, McKay D, Blumberg J. The phytochemical composition and antioxidant actions

of tree nuts. NCBI. 2010;19(1):117–23.

8. Kim Y, Keogh J, Clifton P. Benefits of Nut Consumption on Insulin Resistance and

Cardiovascular Risk Factors: Multiple Potential Mechanisms of Actions. Nutrients.
2017;9(11):1271.
Glucose and Insulin Response to Phytochemical Wright 13

Appendix

Figure 1.1: The means and standard deviations of characteristics of those that participated in the
study

N=7 (n=1 male) (n=6 female)

Height (cm) Weight (kg) BMI (kg/m^2) Age

Mean 158.9 63.6 24.7 29.7

SD 12.0 6.0 2.4 7.7

Glucose and Insulin Response to Phytochemical Wright 14

Figure 1.2: The mean and standard deviation of plasma glucose concentration (mg/dL) at
different time points (minutes) with the consumption of one slice of white bread (WB), 28 g of
dried goji berries (GB), and 28 g of dried goji berries and 30 g of almonds (A)

WB GB A Total

0 minutes 110.4 ± 20.7 104.1 ± 3.5 100.7 ± 5.9 105.1 ± 12.6

15 minutes 115.9 ± 10.6 122.6 ± 8.0* 118.7 ± 13.1 119.0 ± 10.6

30 minutes 124.1 ± 19.7 118.3 ± 17.3 128.0 ± 13.7 * 123.5 ± 16.7

45 minutes 137.4 ± 18.2 GB 112.7 ± 16.8 WB 125.3 ± 14.6 * 125.1 ± 18.9

60 minutes 141.0 ± 20.4 *, GB 115.9 ± 12.3 WB 119.9 ± 11.4 * 125.6 ± 18.3

90 minutes 120.7 ± 11.4 107.0 ± 8.5 110.3 ± 13.7 112.7 ± 12.4

120 minutes 107.1 ± 9.4 108.9 ± 9.4 107.9 ± 6.0 108.0 ± 8.0

* Indicates significant difference compared to baseline within the same trial

WB: Indicates significant difference from white bread at same time point
GB: Indicates significant difference from goji berries at same time point
A: Indicates significant difference from goji berries and almonds at same time point
Glucose and Insulin Response to Phytochemical Wright 15

Figure 1.3: The mean amounts of plasma glucose concentration (mg/dL) at different time points
(minutes) with the consumption of one slice of white bread (WB), 28 g of dried goji berries
(GB), and 28 g of dried goji berries and 30 g of almonds (A)

* Indicates significant difference compared to baseline within the same trial

WB: Indicates significant difference from white bread at same time point
GB: Indicates significant difference from goji berries at same time point
A: Indicates significant difference from goji berries and almonds at same time point
Glucose and Insulin Response to Phytochemical Wright 16

Figure 1.4: The mean and standard deviation of serum insulin concentration (ulU/mg) at different
time points (minutes) with the consumption of one slice of white bread (WB), 28 g of dried goji
berries (GB), and 28 g of dried goji berries and 30 g of almonds (A)

WB GB A Total

0 minutes 3.2 ± 2.4 2.7 ± 2.2 4.3 ± 3.1 3.4 ± 2.5

30 minutes 11.3 ± 5.1 11.4 ± 11.6 11.6 ± 6.7 11.4 ± 7.8

60 minutes 11.9 ± 6.3 4.0 ± 3.6 A 13.6 ± 5.1 *, GB 9.8 ± 6.5

90 minutes 5.4 ± 3.5 5.2 ± 1.9 6.4 ± 3.2 5.7 ± 2.8

120 minutes 4.9 ± 2.3 4.1 ± 2.4 4.9 ± 3.7 4.6 ± 2.7

* Indicates significant difference compared to baseline within the same trial

WB: Indicates significant difference from white bread at same time point
GB: Indicates significant difference from goji berries at same time point
A: Indicates significant difference from goji berries and almonds at same time point
Glucose and Insulin Response to Phytochemical Wright 17

Figure 1.5: The mean amounts of serum insulin concentration (ulU/mL) at different time points
(minutes) with the consumption of one slice of white bread (WB), 28 g of dried goji berries
(GB), and 28 g of dried goji berries and 30 g of almonds (A)

* Indicates significant difference compared to baseline within the same trial

WB: Indicates significant difference from white bread at same time point
GB: Indicates significant difference from goji berries at same time point
A: Indicates significant difference from goji berries and almonds at same time point
Glucose and Insulin Response to Phytochemical Wright 18

Manuscript Rubric
The manuscript should be written in past tense.
Introduction, methods, results, and discussion are continuous. Abstract, references, and
appendix are individual pages.
References are in JAND format
Format: everything double spaced, except abstract