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AOAC First Action Method 2016.02 (Biotin) MLT Report
Page 5 of 16
3.2. MLT Participants
Extensive campaign was carried out through AOAC and ISO/IDF meetings / contacts to recruit potential
laboratories for the multi lab testing. Formal invitation has been sent to more than 30 laboratories around
the world by the study director and 15 laboratories agreed to participate. Delivery of SPIFAN matrices to a
laboratory in Thailand was failed / cancelled due to issues with the recipient’s / country’s import clearance
requirements. Two laboratories could not progress at all with the MLT on time due to inadequate resources
or alternative problems. The details of the remaining 12 laboratories from 10 different countries participated
in the MLT study is given below in Table 2.
Table 2: MLT Participants
Lab # Name of the laboratories
Physical Address
Country
1
Abbott Laboratories
3300 Stelzer Road, Columbus, Ohio 43219
United States
2
Aquanal Laboratoire Aquitaine Analyses 151 bis, Avenue Jean Jaures, Pessac 33600
France
3
AsureQuality New Zealand Limited
131 Boundary Road, Auckland 0600
New Zealand
4
AsureQuality Singapore Pte.
29 Tai Seng Avenue 534119
Singapore
5
DTS Food Laboratories
71 Boundary Road, Melbourne VIC 3051
Australia
6
FirstSource Laboratory Solutions LLP
IDA, Nacharam Cross Rd, Hyderabad-500076 India
7
Fonterra Co-operative Group Limited
Cnr No.1 Rd & SH26, Waitoa 3341
New Zealand
8
Merieux NutriSciences
3600 Eagle Nest Dr, Crete IL 60417
United States
9
R-Biopharm Rhône Ltd
45 Acre Road, Glasgow G20 0XA
Scotland
10
Mead Johnson, China
Xia Yuan Rd, Dongji ID, Guangzhou, 510730
P.R. China
11
Mead Johnson, Netherlands
Middenkampweg2, Nijmegen, 6545CJ
Netherlands
12
Nestle Research Centre, Switzerland
Case Postale 44, Lausanne 26, CH-1000
Switzerland
3.3. Analytical Method
The analytical method provided to the laboratories for the MLT is the same as it is published in Journal of
AOAC Volume 99, Number 4, 2016, pages 1110 to 1112 which is codified as AOAC 2016.02. Extensive
details of the sample preparation, chromatography, calculation, reporting criteria were specified in the
method and the protocol provided to the laboratories. None of the laboratories that participated in the MLT
recorded any modifications or deviations from the documented procedure. This information was requested
to assess the suitability of the method for further approval as Final Action AOAC method.
3.3.1. Title
DETERMINATION OF BIOTIN BY LIQUID CHROMATOGRAPHY COUPLED WITH
IMMUNOAFFINITY COLUMN CLEAN-UP EXTRACTION
3.3.2. Introduction
AsureQuality Auckland Laboratory has initiated a method to facilitate a specific, precise, accurate and
robust procedure for the analysis of biotin from Infant Formula and Adult / Pediatric Nutritional Formulas.
The method also has an assured limit of quantification of 0.1µg/100g (1ppb) based on a simple
mathematical relationship between lowest standard and dilution. The method involves immunoaffinity
column (R-Biopharm Rhone, EASI-EXTRACT biotin column or equivalent) clean-up and extraction followed
by liquid chromatography with UV detection at 200nm.
3.3.3. Principle / Methodology
The sample is dispersed in phosphate buffered saline and autoclaved at 121±2
⁰
C for 25 minutes. The
sample is cooled to room temperature and then diluted to 100mL in a volumetric flask. The extract is
centrifuged and filtered using a Whatman glass microfiber filter paper. Clear filtrate is collected for clean-up
and extraction. Biotin immunoaffinity column is mounted onto a SPE manifold. A disposable syringe barrel
is connected to the immunoaffinity column as a reservoir. The buffer in the affinity column is drained and
the sample filtrate is loaded through the reservoir and allowed to flow through by gravity. The column is
2016.02 (FEBRUARY 2017) BIO-02 MLT REPORT
FOR ERP USE ONLY
DO NOT DISTRIBUTE