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AOAC Final Action Method 2016.02 (Biotin)
Page 2 of 16
In view of the absence of a standard analytical procedure for total biotin, the biotin working group of
SPIFAN activity mooted by AOAC International has developed a Standard Method Performance
Requirements (SMPR 2014.005) in search for a candidate method to be considered as a global dispute
resolution standard.
The AsureQuality Auckland Laboratory has developed a method to facilitate a specific, precise, accurate
and robust procedure for the analysis of biotin and biocytin from Infant Formula and Adult / Pediatric
Nutritional Formulas (1-8). The method meets the SMPR and 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 LC - UV set at 200nm. The laboratory has carried out extensive Single
Laboratory Validation (SLV) study following SPIFAN SLV guidelines (Appendix L: AOAC International)
using SPIFAN kit and NIST SRM 1849a. The SLV along with the method was reviewed by the AOAC
SPIFAN expert review panel (ERP) in 2016 and approved for First Action Official Method of Analysis (OMA)
2016.02.
Collaborative Study (Multi Laboratory Testing)
The objective of the Multi Laboratory Testing (MLT) study was to establish the precision and accuracy of
the method in various laboratories to demonstrate the suitability of the method as an international reference
method with endorsement as AOAC Final Action, and possible adoption by ISO and Codex.
The study was chaired by Dr George Joseph from AsureQuality New Zealand and is monitored by AOAC
International (SPIFAN) Official Method Board through its delegated personnel. The study is divided in two
parts: method set up and qualification of participants (Part 1) and multi laboratory testing by the qualified
participants (Part 2).
Extensive campaign was carried out through AOAC and ISO/IDF meetings / contacts to recruit potential
laboratories for the MLT. 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 within the time frame due to inadequate resources
or alternative problems. The details of the remaining 12 laboratories from 10 different countries participated
in the MLT study (Table 2016.02A)
Table 2016.02A: 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
The study was carried out using SPIFAN II matrices, which represent most of the products in the scope of
the project (Infant Formula and Adult Nutritionals made from any combination of milk, soy, rice, whey,
hydrolysed protein, starch and amino acids, with and without intact protein). Fourteen different product
types were provided to each laboratory which includes two practice samples and 12 samples for multi
laboratory testing over two separate days. Two split samples of each matrix were included in the package
as blind coded duplicates to analyse on the same day. The sample details are summarised in Table
2016.02B
2016.02 (BIOTIN) FINAL ACTION REVIEW
FOR ERP USE ONLY
DO NOT DISTRIBUTE
200