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INFANT FORMULA AND ADULT NUTRITIONALS

Martin & Campos-Gim

É

nez

: J

ournal of

AOAC I

nternational

V

ol.

98, N

o.

6, 2015 

1697

Received May 19, 2015. Accepted by SG June 13, 2015.

1

Corresponding author’s e-mail: esther.campos-gimenez@rdls.

nestle.com

The method was approved by the AOAC

Official Methods Board

as Final Action.

See

“Standards News,” (2014)

Inside Laboratory

Management

, July/August issue.

The AOAC Stakeholder Panel on Infant Formula and Adult

Nutritionals (SPIFAN) invites method users to provide feedback on the

Final Action methods. Feedback from method users will help verify

that the methods are fit for purpose and are critical to gaining global

recognition and acceptance of the methods. Comments can be sent

directly to the corresponding author.

DOI: 10.5740/jaoacint.15-127

Pantothenic Acid (Vitamin B

5

) in Infant Formula and Adult/

Pediatric Nutritional Formula by Ultra-High Pressure Liquid

Chromatography/Tandem Mass Spectrometry Method:

Collaborative Study, Final Action 2012.16

Frederic Martin and Esther Campos-Giménez

1

Nestlé Research Center, Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland

Collaborators: A. Aoude-Werner, S. Bandhari, G. Jaudzems, I. Malaviole, M. Nixon, T. Norloos, L.T. Tanderup, S. Tennyson,

C. Tool, M. Torres, P. van der Burgh, M. Vermeulen, C. Weihong, B. Wu, L.K. Yap

In order to determine repeatability and

reproducibility of AOAC First Action Method 2012.16

[Pantothenic Acid (Vitamin B

5

) in Infant Formula and

Adult/Pediatric Nutritional Formula by Ultra-High

Pressure Liquid Chromatography/Tandem Mass

Spectrometry], a collaborative study was organized.

The study was divided in two parts: method setup

and qualification of participants (part 1) and

collaborative study participation (part 2). For part 1,

each participating laboratory was asked to analyze

two practice samples using the aforementioned

method. Laboratories that provided results within

a range of expected levels were qualified for part 2,

during which each laboratory received 10 samples in

blind duplicates. Results have been compared to the

Standard Method Performance Requirement

(SMPR

®

)

2012.009 established for pantothenic acid. Precision

results (repeatability and reproducibility) were within

the limits stated in the SMPR. Repeatability ranged

from 1.3 to 3.3%, and reproducibility ranged from 4.1

to 7.0%. Horwitz ratio (HorRat) values were all <1,

ranging from 0.33 to 0.69. The AOAC Expert Review

Panel on Stakeholder Panel on Infant Formula and

Adult Nutritionals Nutrient Methods determined that

the data presented met the SMPR and recommended

the method for Final Action status, which was then

granted by the AOAC Official Methods Board.

P

antothenic acid (PA; vitamin B

5

) is commonly present

in foods of either plant or animal origin. This compound

is an essential nutrient for humans, i.e., it is necessary

to synthetize coenzyme-A, which is needed in a vast range of

biological roles, such as metabolism of fatty acids; it also plays

a key part in the Krebs cycle. Historically, determination of PA

was performed with a microbiological assay using

Lactobacillus

plantarum

and its turbidimetric growth (1, 2). Even if this

method is rather sensitive, its specificity is limited when dealing

with complex food matrixes.

More specific techniques have been tested for the analysis

of this compound, such as indirect ELISA (3–5) and

radioimmunoassay (6). Different LC methods have been

developed as well, but the absence of a strong UV chromophore

results in few methods using this detection available for

the analyst. This detection difficulty can be circumvented

by using highly selective MS. Andrieux et al. (7) used this

approach in 2012, with a method combining a rapid sample

preparation prior to the analysis of PA by ultra-high-pressure

liquid chromatography (UHPLC) with a triple quadrupole MS

detection. This method was proposed to the AOAC Stakeholder

Panel on Infant Formula and Adult Nutritionals (SPIFAN) and

was approved as First Action AOAC

2012.16

method (8), with

a recommendation to advance to a multilaboratory collaborative

study. This paper presents the results of that collaborative study.

Method

AOAC First Action Method

2012.16

was used, with minor

modifications, mainly editorial.

AOAC Official Method 2012.16

Pantothenic Acid (Vitamin B

5

)

in Infant Formula and

Adult/Pediatric Nutritional Formula

Ultra-High Pressure LC/MS/MS Method

First Acction 2012

Final Action 2015

ISO–AOAC Method

(Applicable to the determination of free PA in infant formula

and adult/pediatric nutritional formula.)

Caution

: Consult Material Safety Data Sheets prior to using

chemicals and adhere to the safety precautions provided. Wear

personal protective equipment when necessary.

217