AOAC SPIFAN Review Team Meeting Book-July 22, 2015

Method Validation Report

Version

Page

5 of 49

Document Title

Validation Report for the Determination of Thiamin HCl, Riboflavin, Niacin

(Nicotinic Acid and Nicotinamide), Pantothenic acid, Vitamin B6

(Pyridoxine, Pyridoxal and Pyridoxamine) and Biotin in Infant Formula and

Adult Nutritionals by LC-MS/MS

Table 7: Linearity of Pyridoxal

Concentration

Area

Response 1

Area

Response 2

Area

Response 3

(ng/ml)

Standard 1

0.20

0.0305

0.0331

0.0259

Standard 2

0.40

0.0502

0.0547

0.0514

Standard 3

0.81

0.0990

0.1031

0.1080

Standard 4

1.21

0.1575

0.1494

0.1479

Standard 5

1.61

0.1923

0.1976

0.2023

Standard 6

2.02

0.2584

0.2521

0.2513

Standard 7

2.42

0.2915

0.2988

0.2923

Standard 8

4.84

0.5937

0.5826

0.5890

Standard 9

9.68

1.1490

1.1840

1.1950

Standard 10

14.53

1.8090

1.6980

1.7890

Standard 11

19.37

2.3010

2.3860

2.3510

Standard 12

24.21

2.8450

2.8950

2.9310

Standard 13

29.05

l3.2690

3.3140

3.4790

Standard 14

33.9

4.0130

3.9200

3.9570

Standard 15

38.74

4.6240

4.6200

4.7880

Standard 16

43.58

5.2150

5.035

5.2650

Slope

(weighting: 1/x)

0.11825

0.11726

0.1208

Y-Intercept

0.0067

0.00983

0.0037

Correlation

Coefficient

0.99967

0.99971

0.9998

Acceptance

Criteria

Correlation Coefficient (r) ≥ 0.995

Acceptable Y/N

Remark: performed

≥6 point

s standard curve to cover working range of product ranges,

Correlation Coefficient

(r) ≥ 0.995

Depending on matrix and levels of pyridoxal in the product, range of curve can be selected to

decrease number of standards.

BVIT-01 (SLV)

FOR ERP/WORKING GROUP USE ONLY

DO NOT DISTRIBUTE

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