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Fonterra Co-operative Group

Page 5

Table 1: Samples used during method validation

Sample

Code

Fortified

Child Formula Powder

00847RF00

No

Infant Elemental Powder

00796RF

No

Adult Nutritional RTF

(1)

High Protein

00821RF00

No

Adult Nutritional RTF High Fat

00820RF00

No

Infant Formula RTF, Milk-Based

EV4H2Q

No

NIST SRM

(2)

1849a

CLC10-b

D

3

Infant Formula Powder Partially Hydrolysed Milk-Based

410057652Z

D

3

Infant Formula Powder Partially Hydrolysed Soy-Based

410457651Z

D

3

Toddler Formula Powder Milk-Based

4052755861

D

3

Infant Formula Powder Milk-Based

4044755861

D

3

Adult Nutritional Powder Low Fat

00859RF00

D

3

Child Formula Powder

00866RF00

D

3

Infant Elemental Powder

00795RF

D

3

Infant Formula Powder FOS/GOS-Based

50350017W1

D

3

Infant Formula Powder Milk-Based

K16NTAV

D

3

Infant Formula Powder Soy-Based

E10NWZC

D

3

Infant Formula RTF Milk-Based

EV4H2R

D

3

Adult Nutritional RTF High Protein

00730RF00

D

3

Adult Nutritional RTF High Fat

00729RF00

D

3

In-house Reference

D

3

(1)

RTF = Ready-To-Feed

(2)

SRM = Standard Reference Material

Linearity

Within the working range, signal response will have a linear relationship to analyte concentration and the

extent of this range may be demonstrated. Regression calculations on their own are insufficient to establish

linearity and other tests are required to demonstrate linear detector response over the working range. They

can include a visual inspection of the linear regression line and residuals, as well as back calculation of

standard concentrations.

Linearity was evaluated by analysis of 6-level calibration standards on three different days. Regression and

residuals plots of the ratios of the concentrations of analyte vitamin D with stable isotope labelled vitamin D

against the ratios of peak areas of analyte vitamin D with stable isotope labelled vitamin D were prepared.

Linearity for vitamin D

2

and vitamin D

3

was demonstrated by the calculation of correlation coefficients, back-

calculation of standard concentrations (Tables 2

3), and visual evaluation of regression and residuals plots

(Figures 1

6).

The average range for vitamin D

2

in linearity standards was 0.3

59.1 ng mL

-1

, which is equivalent to a

sample powder concentration of 0.3

65 μg hg

-1

(0.04

7.3 μg hg

-1

RTF) which extends beyond both the

VitD-18 (February 2016)

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