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LC Technical Note

Analysis of Vitamin E in food by HPLC

34
GL Sciences Inc.

Chemical Structures
R1

This is an application data of analyzing Vitamin E in food

HO

by HPLC. Vitamin E is a fat-soluble vitamin, which exists


O

R2

tocopherol and tocotrienol. Vitamine E were used as

R3

antioxidant.
Detection was made by Fluorescence detector and
confirmed a good peak shape.
(K. Suzuki)

V.E.

R1

R2

R3

-tocopherol

C H3

C H3

C H3

-tocopherol

C H3

C H3

-tocopherol

C H3

C H3

-tocopherol

C H3

Analysis of Standard Solution

( 25g/L)
( 25g/L)
( 25g/L)
( 25g/L)

1. dl--tocopherol
2. dl--tocopherol
3. dl--tocopherol
4. dl--tocopherol

FLU
4

10
Time (min)

HPLC Conditions
Column
: Inertsil NH2
(5m, 250 x 4.6 mm I.D.)
Col. Temp.
: 40
Detection
: FL Ex 298 nm Em 330 nm
Eluent

6000000

-tocopherol

5000000

R = 0.9999

4000000

B) 2-Propanol

2000000

Flow Rate
: 1.2 mL/min
Injection Vol. : 20L

R = 0.9999

7000000

3000000

A/B/C = 5/6/1000, v/v

-tocopherol

8000000

: A) CH3COOH

C) Hexane

20

9000000

Area

-tocopherol
2

R = 0.9999

1000000

-tocopherol

R = 1.000

100

200

300

400

500

600

Conc.g/L

Calibration Curve

GL SciencesLCTechnicalNote

Sample Preparation Method

Fat

General Foods
Sample

Sample

0.5g
1w/v%NaCl aq. 0.5mL
3w/v% Pyrogallol in Ethanol 10mL
60w/v%KOH aq. 1mL

0.5g

Final Volume

Saponification
Adjust the final volume up to
5mL using Hexane

70, 30min
Cooling
1%NaCl aq. 22.5mL

HPLC-FL
Extraction
Hexane Ethyl acetate9115mL,Shaking5min,
Centrifugation3000rpm, 5min, (Below room temperature)

Liquid-Liquid
Extraction

Lower layer

Supernatant liquid

Extraction
HexaneEthyl acetate 9115mL, Shaking 5min,
Centrifugation 3000rpm, 5min, (Below room temperature)
Supernatant liquid Lower layer
Waste

Concentration
Vacuum concentration, (Below 40)

Final Volume
Adjust the final volume up to
5mL using Hexane

HPLC-FL

GL SciencesLCTechnicalNote

Analysis of Foods

400

200

200

FLU

FLU

400

1. dl--tocopherol
2. dl--tocopherol
3. dl--tocopherol
4. dl--tocopherol

1
4
2

10
Time (min)

20

10
Time (min)

Cooking oil a 10 times dilution

20

Butter
1

200

200

FLU

FLU

400

400

23

3
0

10
Time (min)

20

10
Time (min)

20

Mulukhiya

Broiled eel

1000

800

200

400

100

FLU
600

FLU

200

2
0

23
0

10
Time (min)

20

Almond a 10 times dilution

10
Time (min)

Baby formula

20

GL SciencesLCTechnicalNote

Inert Family
Inertsil

High Performance LC Column

InertCap GC Capillary Column


InertSep Solid Phase Extraction for pretreatment of analysis

Visit our Website on


http://www.glsciences.com/
http://www.inertsil.com/
GL Sciences also offers an extensive range
of GC and LC consumables.

Contact us
GL Sciences, Inc. Japan
22-1 Nishishinjuku 6-chome, Shinjuku-ku, Tokyo, 163-1130 Japan
TEL: +81-3 (5323)6620 FAX: +81-3 (5323)6621

GL Sciences, Inc. USA


4733 Torrance Blvd. Ste 255, Torrance, CA 90503
Tel: (310)265-4424 FAX (310)265-4425

Distributors Outside of Japan and USA


GL Sciences uses distributors in many countries.
You can find a local distributor in your country in the following url.
http://www.glsciences.com/products/contact.html
E-MAIL: world@gls.co.jp