final poster_aprakash

1
Standard Graphs: Introduction Soy is very common in Japanese cuisine. In this experiment, we gathered 28 soy-containing food products from an organic market (located in Miyazaki, Japan) and analyzed the isoflavin concentration through the use of HPLC, and the hydrolysis and non- hydrolysis method. Isoflavin is classified as an aglycon. Aglycon’s are powerful antioxidants present in soy products that help to prevent diseases such as cancer, and inflammatory responses. Conducting this experiment provided the experience needed for quality control, and can also help in new drug development. Because isoflavin is a powerful antioxidant, this method of determining antioxidant levels can be used with other cosmetic active ingredients to determine how effective a prospective product is. Methods & Materials The peaks of the various isoflavones (daidzin genistin, daidzein, glycitein, genistein) were found using the HPLC machine. Isoflavin was extracted using the hydrolysis and non-hydrolysis solvents. Materials used: -Microcrush Machine -Centrifuge + Drying Centrifuge -HPLC -Hydrolysis Solvent ( 50% Phosphate Buffer, 50% MeOH) -Non-Hydrolysis Solvent (50% pure water, 50% MeOH) -Test Tube Heating Block -Pipets +Test Tubes with caps -28 Soy Product Samples -HPLC Standards + Vials -Microsoft Excel Conclusions After analyzing the data obtained from this experiment, it is evident that goziru had highest isoflavin content, and adjusted soy milk had the lowest isoflavin content. This experiment had a duration of 11 weeks and each sample was tested twice using both hydrolysis and non- hydrolysis method. Acknowledgements Contact Information Email: [email protected] u Phone: (440)725-9093 Cosmetic Science Formulation and Design Major Chemistry Minor -Special thanks to Dr. Hiroyuki Sakakibara, Dr. Ming Liu, Hikari Sayaka, Dr. Masahito Suiko, and Ms. Saki Ishikawa, for allowing me to have a great internship experience in Miyazaki, Japan. -Thank you to Dr. Gabriella Baki, Mary Jo Borden, and Dr. Ming Liu for their assistance and guidance throughout the internship process. -A very special thank you to James T. Slama for funding my travel expenses. Determining Isoflavin Concentration of Soy Food Products in Miyazaki, Japan Arushi Prakash ab . Dr. Sakakibara Hiroyuki a , Hikari Sayaka a a University of Miyazaki, Department of Biochemistry b The University of Toledo, College of Pharmacy and Pharmaceutical Sciences Calculations: Standard graphs for daidzein, daidzin, genistein, genistin, glycitein, and glycitin were created on Excel. The equation of the best fit line was then displayed on the graph. HPLC readings for the samples were then inputted into each of the standard equations as the “y” value, and the variable “x” was solved for. If the soy sample is a liquid , the x-value was multiplied by 0.01 to calculate the concentrations of each of the different isoflavin types. If the soy sample had to be weighed using a balance (i.e. Soy Sauce 2 Goziru Goziru 2 Kinako Kinako 2 Kinako 3 Okara Okara 2 Freeze Dr... Freeze Dr... Miso Mix Pure Miso miso Edamame Edamame ... Moyashi Soy Protein Soy Prote... Soykara S... Cold Tofu Film Tofu Soft Tofu Not adjus... Adjusted ... Big Fried... Small Fri... Natto 0.02648 25.585 14.26 7.033 24.393 11.841 22.9 5.09 6.379 3.1304 15.5491 3.1445 3.66 0.1 3.1 7.01 19.668 17.1 22.062 3.495 1.899 1.6 0.1 0.01 12.4697 3.1 2.1 Isoflavin Concentrations In Various Soy Food Products Soy Sample Isoflavin Concentration Total Isoflavin Concentration in µl Of Each Soy Food Product: 0 10 20 30 40 50 60 70 80 90 100 0 10000 20000 30000 40000 50000 60000 70000 f(x) = 646.0232343888 x − 638.472330782 R² = 0.999815547922638 Dasizin 0 10 20 30 40 50 60 70 80 90 100 0 20000 40000 60000 80000 100000 120000 f(x) = 1103.3088742158 x − 1241.3759225233 R² = 0.99975698463799 Genistin 0 10 20 30 40 50 60 70 80 90 100 0 10000 20000 30000 40000 50000 60000 70000 f(x) = 659.0950567426 x − 662.7229621521 R² = 0.999805326738216 Daidzein 0 10 20 30 40 50 60 70 80 90 100 0 20000 40000 60000 80000 100000 f(x) = 848.3782042939 x − 859.5727946877 R² = 0.999804766694499 Glycitein 0 10 20 30 40 50 60 70 80 90 100 0 20000 40000 60000 80000 100000 120000 f(x) = 1080.615109846 x − 1137.803361074 R² = 0.999786464998967 Genistein SCC 70th Annual Scientific Meeting and Technology Showcase - New York City, NY December 10-11, 2015

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Page 1: Final poster_APrakash

Standard Graphs:IntroductionSoy is very common in Japanese cuisine. In this experiment, we gathered 28 soy-containing food products from an organic market (located in Miyazaki, Japan) and analyzed the isoflavin concentration through the use of HPLC, and the hydrolysis and non-hydrolysis method.

Isoflavin is classified as an aglycon. Aglycon’s are powerful antioxidants present in soy products that help to prevent diseases such as cancer, and inflammatory responses. Conducting this experiment provided the experience needed for quality control, and can also help in new drug development. Because isoflavin is a powerful antioxidant, this method of determining antioxidant levels can be used with other cosmetic active ingredients to determine how effective a prospective product is.

Methods & MaterialsThe peaks of the various isoflavones (daidzin genistin, daidzein, glycitein, genistein) were found using the HPLC machine. Isoflavin was extracted using the hydrolysis and non-hydrolysis solvents.

Materials used:-Microcrush Machine-Centrifuge + Drying Centrifuge-HPLC -Hydrolysis Solvent ( 50% Phosphate Buffer, 50% MeOH)-Non-Hydrolysis Solvent (50% pure water, 50% MeOH)-Test Tube Heating Block-Pipets +Test Tubes with caps-28 Soy Product Samples-HPLC Standards + Vials-Microsoft Excel

ConclusionsAfter analyzing the data obtained from this experiment, it is evident that goziru had highest isoflavin content, and adjusted soy milk had the lowest isoflavin content. This experiment had a duration of 11 weeks and each sample was tested twice using both hydrolysis and non-hydrolysis method.

Acknowledgements

Contact InformationEmail: [email protected]: (440)725-9093

Cosmetic Science Formulation and Design MajorChemistry Minor

-Special thanks to Dr. Hiroyuki Sakakibara, Dr. Ming Liu, Hikari Sayaka, Dr. Masahito Suiko, and Ms. Saki Ishikawa, for allowing me to have a great internship experience in Miyazaki, Japan.-Thank you to Dr. Gabriella Baki, Mary Jo Borden, and Dr. Ming Liu for their assistance and guidance throughout the internship process.-A very special thank you to James T. Slama for funding my travel expenses.

Determining Isoflavin Concentration of Soy Food Products in Miyazaki, JapanArushi Prakashab. Dr. Sakakibara Hiroyukia, Hikari Sayakaa

aUniversity of Miyazaki, Department of BiochemistrybThe University of Toledo, College of Pharmacy and Pharmaceutical Sciences

Calculations:

• Standard graphs for daidzein, daidzin, genistein, genistin, glycitein, and glycitin were created on Excel. The equation of the best fit line was then displayed on the graph.

• HPLC readings for the samples were then inputted into each of the standard equations as the “y” value, and the variable “x” was solved for.

• If the soy sample is a liquid , the x-value was multiplied by 0.01 to calculate the concentrations of each of the different isoflavin types.

• If the soy sample had to be weighed using a balance (i.e. is a solid) the x-value was multiplied by 0.01, and then divided by the weight of the product.

Soy

Sauc

e 2

Goz

iru

Goz

iru

2

Kina

ko

Kina

ko 2

Kina

ko 3

Oka

ra

Oka

ra 2

Free

ze D

ried

T...

Free

ze D

ried

...

Mis

o M

ix

Pure

Mis

o

mis

o

Edam

ame

Edam

ame

Leaf

Moy

ashi

Soy

Prot

ein

Soy

Prot

ein

2

Soyk

ara

Snac

k

Cold

Tof

u

Film

Tof

u

Soft

Tof

u

Not

adj

uste

d S.

..

Adju

sted

Soy

milk

Big

Frie

d To

fu

Smal

l Fri

ed T

ofu

Natt

o

0.02

648

25.5

85

14.2

6

7.03

3

24.3

93

11.8

41

22.9

5.09 6.

379

3.13

04

15.5

491

3.14

45

3.66

0.1

3.1

7.01

19.6

68

17.1

22.0

62

3.49

5

1.89

9

1.6

0.1

0.01

12.4

697

3.1

2.1

Isoflavin Concentrations In Various Soy Food Products

Soy Sample

Isofl

avin

Con

cent

ratio

n

Total Isoflavin Concentration in µl Of Each Soy Food Product:

0 10 20 30 40 50 60 70 80 90 1000

10000

20000

30000

40000

50000

60000

70000

f(x) = 646.023234388833 x − 638.472330781977R² = 0.999815547922638

Dasizin

0 10 20 30 40 50 60 70 80 90 1000

20000

40000

60000

80000

100000

120000

f(x) = 1103.30887421575 x − 1241.3759225232R² = 0.99975698463799

Genistin

0 10 20 30 40 50 60 70 80 90 1000

10000

20000

30000

40000

50000

60000

70000 f(x) = 659.095056742566 x − 662.722962152078R² = 0.999805326738216

Daidzein

0 10 20 30 40 50 60 70 80 90 1000

10000 20000 30000 40000 50000 60000 70000 80000 90000

f(x) = 848.378204293874 x − 859.572794687676R² = 0.999804766694499

Glycitein

0 10 20 30 40 50 60 70 80 90 1000

20000

40000

60000

80000

100000

120000

f(x) = 1080.61510984588 x − 1137.80336107389R² = 0.999786464998968

Genistein

SCC 70th Annual Scientific Meeting and Technology Showcase - New York City, NY December 10-11, 2015