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AOAC Final Action Method 2016.02 (Biotin)

Page 14 of 16

Elution of biotin from IAC:

While assisting a participant investigating low biotin results, it became

apparent that the “back flush” technique was inexactly followed, mainly it seems because the exact

description of this technique was not given. So, rather than performing a true “back flush”, the 3mL

methanol eluate was recycled through the IA cartridge three times. It is possible that this may partly explain

slightly low recovery for some samples, given that the IA cartridge may well have retained a small portion of

this eluent fraction / biotin. The details of the back flush using a syringe plunger / reverse pressure is now

detailed in the method.

Syringe Filter:

At least two laboratories experienced interference with chromatography. The investigation

isolated the issue with syringe filters with extractable chromophores. It was advised to test the filters as part

of the reagent blank preparation

Quality Water:

Quality of water

,

especially the water used for the final extraction is very important to get

clean chromatogram. Type 1 water is recommended and this information will be updated in the method.

UPLC Platform

: The availability of stationary phase (solid-core, kinetex phenyl hexyl) in smaller particle

size for UPLC platforms is encouraging. This would significantly reduce analysis time and improve

throughput, my laboratory is doing further work to validate the change.

Calculation errors:

The calculation error is a common problem which was noticed. It was advised to

participants to use reference samples to prompt any issues with calculation before reporting the results.

REFERENCES

1) Bonjour, J. (1991) Biotin. In: Handbook of Vitamins, L J Machlin Ed, Marcel Dekker, Inc., New York, pp

393- 425

(2) Woollard, D.C., & Indyk, H.E. (2013) Biotin Analysis in Dairy Products. In: B Vitamins and Folates:

Chemistry, Analysis, Function and Effects, Ed V R Preedy, RSC Publishing, Lomdon, UK, pp 377- 395

(3) Livaniou, E., Costopoulou, D., Vassiliadou, I., et al. (2000) Journal of Chromatography A. 881, 331-343

(4) Frappier, F. (1993) Biotin: Properties and Determination in Encyclopedia of food sciences; Food

Technology and Nutrition, Academic Press, London, UK, pp 395-399

(5) Lahely, S., Ndaw, S., Arella, F., et al (1999) Food Chemistry. 65, 253-258

(6) IS EN 15607 (2009) Foodstuffs – Determination of D-biotin by HPLC

(7) Holler, U., Wachter, F., Wehrli, C., et al (2006) Journal of Chromatography B. 831, 8-16

(8) Bitsch, R., Salz, I., & Hotzel, D., (1989) International Journal of Vitamins and Nutritional Research. 59,

59-64

(9) AOAC SPIFAN SMPR 2014.005 (2014)

(10) AOAC Official First Action Method 2016.02

(11) AOAC International, International Study Workbook Version 2.1 for Blind (Unpaired) Replicates

(12) Appendix F: Guidelines for SMPR, AOAC International (2012)

(13) Appendix D: Guidelines for collaborative study procedures to validate characteristics of a method of

analysis, AOAC International (2012).

(14) Biotin (Bio-02) SLV Report 14/05V

ACKNOWLEDGEMENTS

I would like to thank the following individuals for their participation in this study and support during the

course of method set up and the MLT study. Special thanks to Simon Bevis and Elaine Marley of R-

Biopharm Rhône Ltd for supporting the program by providing immunoaffinity columns. Thanks to Neil Choi,

DTS Food Laboratories for analysing the SPIFAN matrices using LCMS/MS method and Shellie Stassie,

Covance Laboratories for organising the MLT samples.

1.

Karen Schimpf and Linda Butler Thompson

, Abbott Laboratories, United States

2.

Isabelle Malaviole

, Aquanal Laboratoire Aquitaine Analyses, France

3.

Michael Hodgson,

George Joseph and Ranjani Devi, AsureQuality Limited, New Zealand

4.

Craig Macfadzean, Xinping Huo and Felicia Lim

, AsureQuality Limited, Singapore

5.

Jirong Wang, Trevor Gill and Lee Kennedy,

DTS Food Laboratories, Australia

6.

Sudhakar Yadlapalli

, FirstSource Laboratory Solutions LLP, India

7.

Harvey Indyk, Brendon Gill, Mumtaz Dole and Cesar Yin

, Fonterra Co-operative, New Zealand

2016.02 (BIOTIN) FINAL ACTION REVIEW

FOR ERP USE ONLY

DO NOT DISTRIBUTE

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