Analyzing Extracted Burdock Leaves with Colorimetric Assays Toward the Study of its Efficacy Against

Type 2 Diabetes

Arctium, commonly known as Burdock, is a plant found in Africa, Europe, and Asia. The main use for this plant in this research and experiment is to find a condition in which we can achieve the best yield of antioxidant activity that will allow the neutralization of free stable radical molecules using colorimetric assays. Burdock’s high in antioxidant levels of polyphenol that works as a preventative in cellular damage (1da Silva LM, ET AL. 2012). Previous data and emerging studies have given thought to Burdocks ability to combat Type 2 Diabetes from the extracts of the root (2Ahangarpour A, et. al. 2015). Type 2 Diabetes is the most common form of Diabetes and in which the human body does not use insulin properly and causes the pancreas to make extra insulin. Over time it causes the pancreases to be overwhelmed and in turn renders the pancreas’s ability to function by its inability to keep the glucose levels normal. In the current study, we will explore the benefit of Burdock leaf extract as a source of antioxidant compounds. First, we will collect various Burdock leaf extracts using various conditions (e.g., ethanolic, aqueous extracts, maceration, 40°C, 90°C, microwave extracts); second, we will assess their radical scavenging activity by DPPH and the total phenolic content. Antioxidants have the ability to bond to free radical molecules and inhibit their side effect; therefore, we hypothesize that the ethanoic or aqueous extracts of the burdock leaf may provide a high amount of antioxidants that could be used for the treatment of radical based disorders such as Type 2 Diabetes. Our study is innovative and simple. It will allow us to select the best antioxidant extract condition toward its evaluation in vitro anti-proliferative activity against insulinoma cell line with high glucose and/or high lipid condition that cause excess of oxidative stress and radical species. Alternatively, we will use cell line for culture from a diabetic patient and a normal patient.

INTRODUCTION

METHODS

RESULTS

EXTRACTION • We measure out 20g of burdock leaves • We extracted burdock leaves in 3 solvents; ethanol, water, and a 60:40 ratio of both •  We dissolved the plant extract under 5 conditions; 40°C, 1 day room temperature, 7 day room temperature, 90°C, and microwave • Filtered our dissolved extraction • Evaporated extraction using air

DPPH ASSAY

•  We prepare stocks for the different plant

conditions, DPPH, Gallic Acid, and Vitamin C

•  We plated vertically and Diluted

POLYPHENOL ASSAY •  We prepared stocks for different plant conditions, Gallic Acid, working standards, Folin Ciocalteus,

Reagent,, and Sodium Bicarbonate

DISCUSSION

Initially, the focus for our experiment was based on Leishmania Major, a parasitic infection that causes ulcers. However, we decided to focus on Type 2 Diabetes because in previous studies the root of burdock was said to have the potential to combat this disease. To make our project more unique we decided to test burdock leaves in hopes to find the same outcome.

FUTURE WORK Hopefully we have paved the way for future scientists to further our project and test anti-proliferative activity against a insulinoma cell line from a diabetic patient and a normal patient. We hope that burdock could eventually be made into a drug that is less toxic and safer for patients of all ages to use, and also lower the amount of type 2 diabetes cases in the nation.

ACKNOWLEDGEMENTS

This project is sponsored by National Science Foundation, No. DRL-1322600. “Research reported in this [publication/press release] was supported by the National Institutes on Minority Health and Health Disparities (NIMHD), a component of the National Institutes of Health (NIH) under award number 2G12MD007592.” A special thanks to our scientist Dr. Skouta and Kassandra Borrego, our research assistant.

REFRENCES

Destiny Hernandez1, Lauren Flores2, Isaac Galvez3, Kassandra Borrego4, Dr. Skouta4

1Andres High School 5400 Sun Valley El Paso, TX 79924 2Chapin High School, 7000 Dyer st. El Paso, TX 79904

3Irvin High School 9465 Roanoke Dr. El Paso, TX 79924

4Department of Chemistry, University of Texas at El Paso, El Paso, TX 79968

•  1Department of Pharmacology, D. S. (2013). •  2Health Research Institute, A., Diabetes Research Center, O., Heidari,

Ghaedi, & Taherkhani R.(2015). •  Effects of Hydro-alcoholic Extract from Arctium lappa L. (Burdock) Root

on Gonadotropins, •  Testosterone, and Sperm Count and Viability in Male Mice with

Nicotinamide/ •  Streptozotocin-Induced Type 2 Diabetes. Malays J Med Sci. •  Ethanolic extract of roots from Arctium lappa L. •  accelerates the healing of acetic acid-induced gastric ulcer in rats: •  Involvement of the antioxidant system. Food Chem Toxicol.

y  =  0.0058x  -­‐ 0.068R²  =  0.993

00.10.20.30.40.50.60.70.80.91

1.11.21.31.41.51.61.71.81.92

0 25 50 75 100 125 150 175 200 225 250 275 300

Absorbance  Units

Gallic  Acid  (mg/L)

Folin  Ciocalteau  Gallic  Acid  Equivalent  of  Arctium Extracts

VITAMIN  C

GASTANDARDS

DLI1-­‐2

DLI1-­‐3

DLI1-­‐4

Figure 2: This graph reads from high to low concentrations, it was the highest from the 40° C condition.

Figure 3: This graph reads from high to low concentrations, and it was the highest from the 1 Day condition.

Figure 4: This graph shows that our 3rd plant extract had the highest amount of quenching antioxidants.

Plant Condition Percent Yield % (in grams)

Mass of Extract (in grams)

Mass of Plant (in grams)

DLI 1-2 1.735 0.347 20 DLI 1-3 1.73 0.346 20 DLI 1-4 3.9575 0.7915 20 DLI 1-16 2.5925 0.5185 20 DLI 1-17 1.673 0.3346 20 DLI 1-18 1.613 0.3226 20

-­‐60  

-­‐40  

-­‐20  

0  

20  

40  

60  

80  

100  

120  

1   0.5   0.25   0.125   0.0625   0.03125   0.015625   0.007812   0.003906   0.001953  

DPPH  In

hibition

 %  

Concentration  ug/uL  

Arctium  1  Day  DPPH  Data  30  min.  

Vitamin  C  

DLI  1-­‐16  

DLI  1-­‐17  

DLI  1-­‐18  

-­‐40  

-­‐20  

0  

20  

40  

60  

80  

100  

120  

1   0.5   0.25   0.125   0.0625   0.03125   0.015625   0.007812   0.003906   0.001953  

DPPH  In

hibition

 %  

Concentration  ug/uL  

Arctium  40  C  DPPH  Data  30  min.  

Vitamin  C  

DLI  1-­‐2  

DLI  1-­‐3  

DLI  1-­‐4  

Figure 1: When the antioxidant is going to neutralize the free radical, there will be a color change from purple to yellow.