AOAC SPIFAN Review Team Meeting Book-July 22, 2015

Method Validation Report

Version

Page

2 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 2: Linearity of Riboflavin

Concentration

Area

Response 1

Area

Response 2

Area

Response 3

(ng/ml)

Standard 1

19.4

0.1026

0.1017

0.1031

Standard 2

38.81

0.2029

0.2048

0.2000

Standard 3

77.62

0.3873

0.4025

0.3942

Standard 4

116.43

0.5952

0.5915

0.5921

Standard 5

155.24

0.7986

0.8112

0.7931

Standard 6

194.04

1.0210

1.0290

1.0050

Standard 7

232.85

1.1850

1.1960

1.1930

Standard 8

271.66

1.4050

1.4130

1.3720

Standard 9

310.47

1.5950

1.6100

1.5690

Standard 10

349.28

1.7690

1.7930

1.7550

Slope

(weighting: 1/x)

0.00512

0.00517

0.00506

Y-Intercept

0.00249

0.002

0.00495

Correlation

Coefficient

0.99985

0.99988

0.99993

Acceptance

Criteria

Correlation Coefficient (r) ≥ 0.995

Acceptable Y/N

Table 3: Linearity of Nicotinic acid

Concentration

Area

Response 1

Area

Response 2

Area

Response 3

(ng/ml)

Standard 1

8.14

0.3067

0.3010

0.3089

Standard 2

16.29

0.5757

0.5779

0.5718

Standard 3

24.43

0.8474

0.8177

0.8243

Standard 4

32.57

1.1120

1.0640

1.0900

Standard 5

40.72

1.3620

1.3500

1.2980

Standard 6

48.86

1.6450

1.6380

1.5870

Standard 7

57.01

1.7620

1.7480

1.7640

Standard 8

65.15

2.0370

2.0660

2.0000

Standard 9

73.29

2.2700

2.2430

2.2480

Slope

(weighting: 1/x)

0.03069

0.0306

0.03001

BVIT-01 (SLV)

FOR ERP/WORKING GROUP USE ONLY

DO NOT DISTRIBUTE

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