OMB Meeting Book_9-11-14

Vitamin A & E - MLT For ERP Use Only DO NOT DISTRIBUTE

SIMULTANEOUS DETERMINATION OF 13-CIS AND ALL-TRANS VITAMIN A PALMITATE (RETINYL PALMITATE), VITAMIN A ACETATE (RETINYL ACETATE), AND TOTAL VITAMIN E (Α-TOCOPHEROL AND DL-Α-TOCOPHEROL ACETATE) IN INFANT FORMULA AND ADULT NUTRITIONALS BY NORMAL PHASE HPLC

CODE:

DATE:

PAGE:

VER:

Purpose:

AOAC 2012.10

Jan 2014

1 of 11

04

AOAC MLT Study

Page: 8 of 11

12.0

1.5

5

95

14.0

1.5

5

95

15.0

1.5

95

5

20.0

1.5

95

5

Note : The gradient given can be altered as required to maximize the analytical separation and avoid interferences. This should only be done by an authorized analyst.

N. SYSTEM SUITABILITY

a. The correlation coefficient of each calibration should be not less than 0.995. b. The resolution between cis and trans forms of Vitamin A Palmitate and 13- cis and trans forms of Vitamin A Acetate should be not less than 1.5. Resolution of a minor cis peak (9 or 11-cis) between the 13-cis and trans peak gives confidence that the resolution is sufficient. c. When stable baseline is obtained, inject a medium reference standard three times in succession and determine the %RSD, The %RSD should be not more than 2%. d. The appropriate number of samples between successive calibrations is typically 6 samples.

e. The target limits of tailing factor for meeting the system suitability are 0.8 to 1.2.

O. CALCULATIONS

The HPLC software will automatically calculate the concentration of the sample in µg/100g for Vitamin A and mg/100g for Vitamin E, (powder or liquid), provided the concentration of the working standards in µg/mL (Vitamin A) or mg/mL (Vitamin E), sample weight / powder equivalent (g), dilution (10mL) and conversion to 100g are entered correctly.

I A )

(



100

S

w



10

 

S

m

s

Where: A S

is the peak area or height of the Vit A or Vit E in the test sample solution

I

is the intercept of the calibration curve is the slope of the calibration curve

S

10 is the volume (mL) of isooctane used 100 is the factor to convert in 100 g basis m S is the sample weight (liquid samples) / powder equivalent in grams (powder samples)

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