International Standards for Food Authenticity and Allergen Detection from ISO TC 34/SC 16 

Horizontal Methods for Molecular Biomarker Analysis

Michael Sussman

Corresponding Author

Michael Sussman, michael.sussman@usda.gov

USDA

Abstract

ISO Technical Committee 34 “Food Products”/Subcommittee 16 “Horizontal methods for molecular 

biomarker analysis” works to ensure that standardized biomolecular testing and laboratory criteria are 

reproducible and technically sound reducing potential disputes between exporting and importing nations 

and increasing predictability in world trade. Harmonized, easy to handle methods of analysis with defined 

patterns and known nomenclatures bring more customers to the market. SC 16/TC 34 has increased 

international stakeholders’ participation in standardizing biomarker testing, improved the quality and 

relevance of these standards and continues to increase transparency in international markets, particularly 

for food authenticity, varietal identification and genetically engineered products. ISO standards have been 

adopted by Codex Alimentarius and many governments throughout the world. The International 

Organization for Standardization (ISO.org) was formed in 1946. It is an independent, non-governmental 

voluntary consensus standard body based in Geneva, Switzerland with a membership of 163 national 

standards bodies. The US ISO member is the American National Standards Institute (ANSI.org) a 

consortium of US standardization organizations. ISO TC 34/SC 16 was created in 2008. There are 42 

participating countries. Its scope is, Standardization of biomolecular testing methods applied to foods, 

feeds, seeds and other propagules of food and feed crops. The US delegation responsible for developing 

the US position for standards development in food authenticity and allergen detection is called the US 

Technical Advisory Group (TAG). It was delegated to the American Oil Chemist’s Society (AOCS.org) by 

ANSI. AOCS also hosts the TC 34/SC 16 international secretariat. TC 34/SC 16 has published 20 standards 

with another 16 under development. Recently published standards include: Technical Specification ISO/TS 

16393 Molecular biomarker analysis — Determination of the performance characteristics of qualitative 

measurement methods and validation of methods, International Standard ISO 16578 Molecular 

biomarker analysis —General definitions and requirements for microarray detection of specific nucleic 

acid sequences and International Standard ISO 20813 Molecular biomarker analysis — Methods of 

analysis for the detection and identification of animal species in foods and food products (nucleic acidbased methods) — General requirements and definitions. More on food authenticity and allergen 

detection work in ISO TC 34/SC 16 is provided.

Poster Category: Authenticity, Fraud, and Adulteration

Poster Number: AUTH-01
Food safety and food authenticity by peptide mass spectrometry – Constitution of new § 64 

LFGB working groups for method validation and standardization

Manfred Stoyke and Rene Becker

Corresponding Author

Manfred Stoyke, manfred.stoyke@bvl.bund.de

Federal Office of Consumer Protection and Food Safety

Abstract

The monitoring of food authenticity and food allergens is a crucial task for consumer protection. In annex 

II of the EU regulation 1169/2011, 12 groups of materials causing most intolerances or allergies in food 

are listed that have to be labelled on products for the final consumer. The testing for food authenticity 

as well as food allergens is mostly done by ELISA or PCR. They also have some limitations and drawbacks. 

ELISA methods often have specificity issues whereas in PCR methods, allergen detection is achieved only 

indirectly via DNA. Moreover, both technologies have limitations in terms of the number of species or 

allergens that can be analysed in the same sample in one assay. For ELISA, typically only one species or 

allergen can be analysed, whereas for PCR, most assays range between one and four allergens. In recent 

years, new methods for food authenticity have been developed utilising liquid chromatography coupled 

to mass spectrometry. Since most food allergies are caused by allergenic proteins, the new methods have 

the potential to detect not only the allergenic ingredient but also the specific protein that causes an 

allergy. Another major advantage of such methods is the detection of a multitude of allergen-causing 

proteins in a single analysis. At present, however, these methods generally lack validation let alone 

standardisation. The Federal Office of Consumer Protection and Food Safety (BVL) in Germany has 

recognized the potential of such novel technology applications and therefore constituted working groups 

with the aim to identify appropriate technologies and validate these for official food control. The new 

working groups, consisting of experts from the field, will focus mainly on the validation and 

standardisation of methods for allergen detection and food authenticity. The poster will provide an 

overview over the current and future work of the newly-formed working groups.

Poster Category: Authenticity, Fraud, and Adulteration

Poster Number: AUTH-02

Coauthors

Rene Becker, Federal Office of Consumer Protection and Food Safety
A traceable two-dimensional image analysis method for the characterizing quality parameters 

in rice-based candidate reference material

Bryan Calderón-Jiménez, Dionisio Gutiérrez-Fallas, Ernesto Montero-Zeledon, Gabriel Molina-Castro, 

and Katia Rosales-Ovares

Corresponding Author

Bryan Calderón-Jiménez, bcalderon@lacomet.go.cr

Chemical Metrology Division, Costa Rican Metrology Laboratory (LACOMET), San José, CR

Abstract

The following study describes the preliminary results of the implementation of two-dimensional image 

analysis as a metrological traceable method for the characterization of a candidate reference material 

(RM) in rice (Oryza sativa L.). Parameters evaluated in the candidate RM were dimensional, physical 

parameters of quality used by national and international regulations to categorize and determine the 

content of the whole kernel, broken kernel and chips in rice. Specifically, a RM was simulated by 

gravimetric mixing of defined portions of whole rice (100 %) and small broken rice (100 %) to obtain a 

specific percentage of whole/broken rice. The proposed method consists of three general processes i) 

sample preparation, ii) capture and image digitization, iii) data processing and analysis. For sample 

preparation, a representative mass of rice was carefully dispersed in an optical work area while avoiding 

the occurrence of grain agglomeration. The capture and digitalization process was carried out with a 

DigiEye system (VeriVide). Metrological traceability was accomplished using an artifact previously 

calibrated and incorporated into the image digitalization of all samples. ImageJ software was used to 

investigate grain parameters which include contrast, brightness, outline, as well as dimensional 

parameters of the grains such as area, perimeter and maximum length. Dimensional parameters data 

were analyzed using RStudio in order to perform graphical and statistical analysis of the results. The 

preliminary results demonstrate the applicability of this method to evaluate dimensional specifications of 

rice that may be subject to international trade. Also, this method provides metrological traceability to the 

meter unit, improving the limitation of classical methods which are traceable to mass. Efforts are currently 

underway to establish this method for the characterization of RM for industrial quality control 

measurements and organization of proficiency testing (PT).

Poster Category: Authenticity, Fraud, and Adulteration

Poster Number: AUTH-04

Coauthors

Dionisio Gutiérrez-Fallas, Physics School, Costa Rica Institute of Technology, Cartago, CR

Ernesto Montero-Zeledon, Physics School, Costa Rica Institute of Technology, Cartago, CR

Gabriel Molina-Castro, Chemical Metrology Division, LACOMET, San José, CR

Katia Rosales-Ovares, Chemical Metrology Division, LACOMET, San José, CR
Meat Authenticity: Does more frequent PT participation improve PT performance?

Heather Jordan and Dr. Matthew Whetton

Corresponding Author

Heather Jordan, Heather.Jordan@LGCGroup.com

LGC Group

Abstract

LGC is a global provider of proficiency testing (PT) schemes in the field of food quality testing. When the 

horse meat scandal struck Europe in 2013, LGC launched a meat authenticity PT scheme within months. 

Since then, additional authenticity test materials have been added, including cheese and fish. Estimates 

from the Grocery Manufacturers Association indicate that food fraud may cost the world economy almost 

$50 billion per year and that 10% of all food products are likely to be adulterated. While not all food 

authenticity issues present a health risk to consumers, most will have an impact on brand reputation and 

consumer confidence. The evolution of molecular methods allows for more frequent and faster analysis

of food materials and; therefore, gives the potential for improve transparency in the industry. As we rely 

on laboratory analytical measurement results to ensure integrity of the food market, we must 

simultaneously and continuously ensure laboratories entrusted with this analysis are providing accurate 

measurement results, using robust and reliable methods. One means, arguably the only means, of 

externally monitoring laboratory performance is through routine proficiency testing. Many requirements 

and guidance for laboratory analysis in the food industry (e.g., some accrediting bodies, a GFSI recognized 

certification program, FSIS guidance documents) set a minimum PT participation of once per calendar 

year, per method or test. As ISO/IEC 17025 moves towards a risk-based approach to determining PT 

frequency, laboratories may wonder how to optimize PT participation. Using data from the LGC meat 

authenticity program over the last two years, this poster will demonstrate that laboratories participating 

two times or more per year achieve higher rates of satisfactory performance than those participating only 

once per year.

Poster Category: Authenticity, Fraud, and Adulteration

Poster Number: AUTH-05

Coauthors

Dr. Matthew Whetton, LGC Group
A Rapid, Univariate FT-NIR Procedure to Determine Moisture Concentration, a Quality 

Parameter, in Olive Oil

Magdi Mossoba, Ali Reza Fardin-Kia, Sanjeewa R. Karunathilaka, Betsy Jean Yakes, and Zachary Ellsworth

Corresponding Author

Magdi Mossoba, Magdi.mossoba@fda.hhs.gov

FDA

Abstract

We recently observed that the weak near-infrared (NIR) band near 5260 cm-1 was relatively more intense 

for extra virgin olive oil (EVOO) than for refined olive oil (ROO). We also observed that its intensity was 

diminished upon heating and erroneously presumed that it may be attributed to volatile carbonyl 

components in EVOO. In the present study we demonstrate for the first time that this band is primarily 

attributed to a water O-H combination band. To accurately determine the intensity of this weak band, 

observed on a shifted and sloping baseline, we measured the peak-to-peak (p-p) height of its first 

derivative. An exponential calibration curve for p-p height versus gravimetrically-determined 

concentration of spiked water was satisfactorily generated. The calibration curve was first evaluated by 

using independent sets of gravimetrically prepared test samples. Subsequently, it was used to determine 

the moisture content, a quality parameter, for a limited set of authenticated reference olive oils whose 

quality and purity were confirmed by official methods. These concentrations, 0.098-0.12% H2O (w/w) for 

EVOO, 0.022-0.030% H2O (w/w) for ROO, and 0.028-0.054% H2O (w/w) for pomace olive oil (POO), were 

consistent with those reported in the literature. The lowest moisture concentration obtained for EVOO 

was subsequently used to segregate a limited set of 88 retail products labeled as EVOO: 42% of these 

products contained 0.098-0.13% H2O (w/w), while the remaining products (58%) yielded lower 

concentrations between 0.026 and 0.097% H2O (w/w). When the purity of these 88 products were 

determined with official methods, only 3% were found to be adulterated. By contrast, the correlation 

between moisture content and other olive oil quality parameters has been reported in the literature and 

has yet to be further investigated and verified based on official methods.

Poster Category: Authenticity, Fraud, and Adulteration

Poster Number: AUTH-06

Coauthors

Ali Reza Fardin-Kia, FDA

Sanjeewa R. Karunathilaka, FDA

Betsy Jean Yakes, FDA

Zachary Ellsworth, University of Maryland, Joint Institute for Food Safety and Applied Nutrition
Analysis of Acylglycerols in Edible Oils by Gas Chromatography Using a Unique Stationary 

Phase

Joseph Konschnik, Colton Myers, Kristi Sellers, Jana Rousova, Shawn Reese, Jaap de Zeeuw, and Chris 

Rattray

Corresponding Author

Joseph Konschnik, joe.konschnik@restek.com

RESTEK Corporation

Abstract

Characterization of edible oils is essential to the food industry due to the amount of fraudulent activity 

that surrounds these products. Some edible oils (e.g. Extra Virgin Olive Oil) carry high value, therefore 

making it an easy target by frauds. By mixing different vegetable oils (e.g. rapeseed, sunflower, etc.) with 

high quality olive oils, manufactures increase their oil yields and make larger profits on these counterfeit 

olive oils. For these reasons, it is important to obtain a triacylglycerol (TAG) fingerprint of edible oils to 

know that they have not been adulterated with other oils. In addition, the freshness of oils can be 

determined by looking at the ration of 1,2 to 1,3-diacylglycerols (DAG). By using a unique gas 

chromatography (GC) stationary phase without bleed interference and retention time shifting due to 

phase lost, and is able to resolve TAGs and DAGs, a full analysis of the edible oil can be conducted for oil 

adulteration and degradation. The analysis and results for these oils will be presented along with and 

examination of column bleed at high GC operating temperatures.

Poster Category: Authenticity, Fraud, and Adulteration

Poster Number: AUTH-07

Coauthors

Colton Myers, RESTEK Corporation

Kristi Sellers, RESTEK Corporation

Jana Rousova, RESTEK Corporation

Shawn Reese, RESTEK Corporation

Jaap de Zeeuw, RESTEK Corporation

Chris Rattray, RESTEK Corporation
Real-Time Authentication of Whiskeys Using DART-QDa Analysis

Emily Britton, Sara Stead, and Renata Jandovia

Corresponding Author

Emily Britton, emily_britton@waters.com

Waters

Abstract

Whiskey, a popular spirit-based drink, is sold as the product of one distillery or as a blend available 

worldwide. Analytical methods are required for process control and quality assurance but production is 

often also strongly regulated with regards to the authenticity of the product. The high popularity of these 

products generates a high risk of fraud. The characteristics of whiskey are strongly influenced by the 

cereals used in fermentation and by distillation, maturation and blending regimes. This leads to 

characteristic profiles exploited for authenticity analysis typically using gas chromatography combined 

with chemometrics. Such methods can be time-consuming, due to the demands of the chromatographic 

separation so there is a need for development of tools for more rapid testing. We have demonstrated the 

potential of Direct Analysis in Real Time (DART) coupled with simple mass detection (QDa) and 

multivariate statistics for the rapid analysis of whisky samples. Here we report further results of 

comparative analysis of authentic samples of whiskey brands and determine whether it is possible to 

distinguish them using LiveID software. LiveID has a visually modern, attractive, web-based interface that 

offers a workflow-driven process that is easy to learn and use. Initially, authentic reference samples are 

used to create and validate a statistical model, which are tested with unknown samples to generate live 

classifications. The output is a simple to interpret yes/no answer delivered in real-time.

Poster Category: Authenticity, Fraud, and Adulteration

Poster Number: AUTH-08

Coauthors

Sara Stead, Waters

Renata Jandovia, Waters