217 / 258
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.comThe 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