AOAC India Section Collaborative Study

1. Single-Laboratory Validation (SLV) and
Multi-Laboratory Testing (MLT): AOAC
International - India Section

2. Objective
One of the prime objective of the India Section of
AOAC International is to apply the global methods (e.g.
AOAC methods) for priority nutrients to Indian foods/food
product matrices of Infant Formula and Adult Nutritionals so
as to build a scientific justification to persuade FSSAI/BIS
and other national regulatory bodies to replace the current
conventional methodologies with these highly accurate
ones as Dispute Resolution Methods.

3. Vitamin B12, like the other B vitamins, is important for metabolism. It
helps in the formation of red blood cells and in the maintenance of the
central nervous system. RDA of Vitamin B12 for Indians (μg/Day)
[ICMR, 2010]: Adult male & Adult female (NPNL)(1.0), Pregnancy (1.2),
Lactation (1.5), Infant <12mths (0.2), Children 1 – 17yrs (0.2 – 1.0)
All the vitamin B12 found in nature is synthesized by microorganisms.
The vitamin is absent in plant foods except when contaminated with
microbes. Liver, meat, egg and milk are good sources.
Traditional microbiological methods for vitamin B12 are not precise and
accurate enough to meet the nutrient specification/claim requirements
of infant, pediatric, and adult nutritionals.
In September 2014, AOAC SPIFAN Nutrients Expert Review Panel
(ERP) have recommended AOAC 2011.10 (“Determination of Vitamin
B12 in Infant Formula and Adult Nutritionals by HPLC”) for Final Action
Official Method status post review of multi-laboratory testing (MLT)
data.

4. Time, min Valve Configuration
0.00–10.5 Configuration 1
10.5–14.5 Configuration 2
14.5–30.0 Configuration 1

5. Applicability
Applicable to the determination of vitamin B12 in infant formula and adult nutritionals. The
method was validated for Stakeholder Panel on Infant Formula and Adult Nutritionals
(SPIFAN) matrices:
1. SRM 1849 (NIST)
2. Infant Formula Powder Milk-Based
3. Infant Formula Ready to Feed Milk-Based
4. Infant Formula Powder Soy-Based
5. Infant Formula Powder Hydrolysate Milk-Based
6. Infant Formula Powder Hydrolysate Soy-Based
7. Infant Formula Powder Elemental (amino acid-based)
8. Child Formula Powder
9. Adult Nutritional Powder
10. Adult Nutritional Powder Low Fat
11. Adult Nutritional RTF High Protein
12. Adult Nutritional RTF High Fat
With all matrixes tested, the method had an average overall intermediate precision of
6.64% RSD, an estimated quantitation limit of 0.8 μg/kg, and a detection limit of 0.2 μg/kg
in prepared samples. The standard range of the method is 2 to 200 μg/L, which
corresponds to an analytical range of 0.8 to 500 μg/kg.

6.  Identification of interested participants (industries/food
manufacturing organizations & analytical test labs)
 Finalization of Indian food product Matrices for SLV (the
same will be used subsequently for MLT)
 Review with AOAC International to see if these Indian
food product Matrices fall under the scope of the current
methodology/applicability
 If the majority of the Indian food product Matrices falls
under the scope of the existing
methodology/applicability, proceed directly to Multi-
Laboratory Testing (MLT) or else proceed to Single Lab
Validation (SLV)

7. Single Lab Validation (SLV)
 The participating industries (whose product falls under the
category of “Infant Formula and Adult Nutritionals” and wish to
include their product in this study) forms a consortium under India
Section of AOAC International to provide samples (for SLV &
MLT) and funding (for SLV and for material handling/logistics
during MLT). This consortium will be headed by the Study
Director who will be responsible for the disbursement of the
samples (for SLV & MLT) . All the financial part will be handled by
the Treasurer of India Section of AOAC International.
 One of independent participating analytical test lab volunteers to
undertake SLV on no-profit, no-loss principle. Industries/food
manufacturing organizations won’t be allowed to undertake this
SLV activity to protect potential conflict of interest.

8.  Required samples and adequate funding provided to the volunteering
analytical test lab by the consortium.
 Volunteering analytical test lab performs SLV as per AOAC guidelines
(Appendix K) and submit the data to AOAC International - India Section
Expert Review Panel (ERP)
 ERP reviews the data and recommends it for MLT.
Multi-Laboratory Testing (MLT)
 Finalization of MLT study protocol
 Participants# volunteer and demonstrate capability on test sample
 Successful participants# take part in full MLT using above food product
matrices
 Study Director collect data and submit to ERP, India Section of AOAC
International
 AOAC India proposes the method to FSSAI/BIS and other national regulatory
bodies as Dispute Resolution Methods
#All the allied cost of MLT needs to be borne by the participants