MONITORING QUALITY TRAITS OF FOOD INGREDIENTS AND DETECTION OF MICROBIAL/CHEMICAL ADULTERANTS/HAZARDS BY VIBRATIONAL SPECTROSCOPY TECHNOLOGIES

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1003936
• Project No.: OHO01339
• Project Start Date: May 4, 2015
• Project End Date: Sep 30, 2019

Project Director
Rodriguez-Saona, LU, EN.

Recipient Organization
OHIO STATE UNIVERSITY
1680 MADISON AVENUE
WOOSTER,OH 44691

Performing Department
Food Science & Technology

Non Technical Summary
Recent safety scares with imported products have raised significant concerns in the U.S. about threats to the 1.2 trillion dollar food industry and to the billions of dollars of imported foods entering the U.S. each year. In addition, quality control is essential in the food industry and efficient quality assurance is becoming critical. Cutting edge sensor technologies can provide a valuable window into in-process food manufacturing to permit optimization of production rate, quality and safety of many food products. Current methods for testing are time-consuming, expensive, labor-intensive, requiring complex procedures of sample pretreatment and well-trained technicians to operate the instrumentation. Thus, it is of paramount importance to develop simpler, quicker, sensitive, and cost-effective methods for monitoring the quality and safety of our foods. We propose to establish reliable quality and safety programs for high-risk ingredients using vibrational spectroscopic techniques such as FT-IR and Raman spectroscopy. The proposed technology would enable detection and characterization of important quality traits and potential microbial and chemical food contaminants via unique spectral signature profiles, permitting real-time and field-based control of the product stream to enhance quality, safety and brand equity. Implementation of rapid testing by the industry and regulatory agencies can streamline food safety and quality assurance and will curtail the growing danger to the health of consumers from adulterated or substituted products.

Animal Health Component

0%

Research Effort Categories

Basic

30%

Applied

60%

Developmental

10%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
711	5010	2000	50%
701	5010	2000	50%

Knowledge Area
711 - Ensure Food Products Free of Harmful Chemicals, Including Residues from Agricultural and Other Sources; 701 - Nutrient Composition of Food;

Subject Of Investigation
5010 - Food;

Field Of Science
2000 - Chemistry;

Keywords

vibrational spectroscopy

quality assurance

food safety

pattern recognition

handheld/portable technology

Goals / Objectives
We propose to establish reliable technologies for monitoring quality traits and detect and quantify potential microbial/chemical hazards using vibrational spectroscopic techniques, including FT-IR and Raman spectroscopy. The objectives of our proposed research focus on developing:Validated prediction models generated with customized vibrational spectroscopy technology to the specifications required by the industry for multiple quality traits.Develop spectral libraries for threat materials and fingerprinting microbial and chemical hazards in food matrices.Our proposed research will focus on mature spectroscopic sensor technologies, to enable pathogen identification with analytical precision equivalent to bench-top instruments found in the laboratory. We will also use our find ways to science and engineering expertise to improve or expand upon these existing technologies to meet challenges specific to food monitoring (e.g., front-end sampling, purpose-built sensor systems and detection algorithms, spectral libraries).Successful development of such real-time and field-based spectroscopic systems will provide significant advantages for food safety monitoring. These technologies can enable greater control of the high volume raw material stream, addressing both safety and brand equity. Implementation by the industry and regulatory agencies of rapid testing procedures based on these technologies would help to streamline food safety and quality assurance and will prevent the growing danger to the health of consumers from adulterated or substituted products as evidenced by the melamine incident. Development of spectral libraries to fingerprint high-risk fresh food crops (i.e. spinach, alfalfa sprouts, and tomatoes) and ingredients (i.e. powdered milk, corn meal, cocoa & peanut butter, fruit juices) would provide the food industry unique detection capabilities to prevent fraud and ensure consumer product safety. Testing done as close to the original source would permit detecting adulteration before an ingredient has been diluted or combined with other ingredients. Vibrational sensing technologies would permit interrogation of produce wash water for chemicals and pathogens to evaluate exposure.

Project Methods
The experimental design will include establishing a comprehensive reference spectroscopic profile for materials/ingredients, model design and analysis of samples using FT-IR, and Raman spectroscopy. We will identify an appropriate sample set to establish a reference spectral range for materials from identified manufacturers. The pure samples will be combined to form a reference spectral range. These samples will be analyzed using FT-IR and Raman spectroscopy, and the spectra will be evaluated for highly specific chemical signatures to characterize particular component make-up.Monitoring quality traits: We will work with economically relevant products (ie. tomato juice, vegetable oils, cocoa butter, among others). Spectra of samples will be collected by using different infrared accessories (transmittance, ATR, diffuse reflectance). Important quality traits will be determined by official reference methods. All spectral data will be analyzed by chemometrics using Partial Least Squares Regression. Multivariate analysis will model relationships between the infrared spectra and quality parameters and its predictive accuracy will be examined with an independent validation set and the accuracy of the models determined.Method sensitivity and quantification microbial/chemical hazards: We will develop robust and sensitive protocols that will allow the rapid quantitation of chemical contaminants and pathogens by Infrared/Raman spectroscopy to yield specific signal of marker compounds. For microbial detection, we will use immunomagnetic separation for target pathogens in high-risk produce contaminated at different levels (10, 102, 103, and 104 CFU/g) to determine level of detection. The magnetic affinity-captured microorganism are applied onto an array slide and analyzed by reflectance Infrared/Raman spectroscopy. For each target pathogen, triplicate samples will be prepared at each level of contamination to evaluate reproducibility. The experiment will be conducted on different days (at least 4 days) to evaluate the robustness of the spectral data. The vibrational spectroscopy technique will provide a tool to current rate-limiting and time-consuming methods to effectively evaluate the safety of produce/ingredients. Furthermore, we expect that the low sensitivity and high accuracy and reproducibility of the IRMS method will provide levels of detection <10 CFU/g. Chemical Contaminants: Uncontaminated high-risk samples (ie. powdered milk, cocoa & peanut butter, fruit juices) will be spiked with different adulterant-like chemicals. It is important that the samples be homogenous so the samples will be thoroughly mixed before analyzing. The model design will be based on two factors: adulterant and concentration. The spiked samples will be analyzed using FT-IR and Raman spectroscopy. Spectral analysis will be carried out by supervised chemometric methods for sample classification (SIMCA) and quantitative (PLSR) analysis. SIMCA consists of assigning training data sets to classes and then a principal component model is created for each class with different confidence regions. Probability clouds (95% CI) are built around the clusters based on PCA scores, allowing SIMCA to be used as a predictive modeling system. This step also involves determining the lower limits of adulterant concentration that each technology can detect. For each chemical entity, triplicate samples will be prepared at each level of contamination. Quantitative models based on infrared spectra will be generated by Partial Least Squares regression (PLSR) using the spiked levels confirmed by using a reference method. Using published data on fundamental vibrations of specific functional groups and available standards will correlate the IR spectral signals. Classification and regression models will be used to generate prediction models and the accuracy and ability of these models will be examined with an independent test set representative of the classes modeled with the training set. Blind samples (the researcher will not have access to its identity before prediction) will be included to test the ability of the models to predict the identity and levels of a potential chemical contaminant.Deployability of protocols/models: We will consider existing technologies such as newly developed portable optical systems for chemical identification which bring the analytical precision of spectroscopy to field applications with spectral resolution equivalent to bench-top instruments. We will also find ways to improve or expand upon existing technologies (front-end sampling, purpose-built sensor systems and detection algorithms, spectral libraries) using our expertise to make the technologies more applicable.

Progress 05/04/15 to 09/30/19

Outputs
Target Audience:The results from our Hatch project were disseminated internationally and nationally through events that target scientific and industrial audiences interested in rapid alternatives for monitoring quality parameters and detection of food contaminants. These events provided a perfect setting for exchanging ideas with experts in the field, discuss current work, receive feedback from potential end-users, develop professional and industrial partnerships and disseminate knowledge through multidisciplinary collaborations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Our Lab has trained 7 OSU Food Science undergraduate students, Sarah Gray, Rachel Larue, Chris Huang, Anna Schmenck, Regina Tanuwidjaja, Kuanrong Zhu and Wan Xin Tan. The students are working on authentication of maple syrup by Raman and sucrose level monitoring in breakfast cereals, respectively. Students are developing skills in spectral collection, chromatographic analysis and data analysis. We graduated 2 PhD and 3 MS students in Food Science at The Ohio State University with dissertation titles: "The New Generation of Handheld Vibrational Spectroscopy Devices -Applications for Authentication of High Valued Commodities" (Dr. Shotts), "Verification of Ingredient Labels in High-Risk Oils and Fruit Juices by Using Vibrational Spectroscopy Combined with Pattern Recognition Analysis" (Dr. Aykas), "Screening for Acrylamide Levels in French Fries Using Portable Vibrational Spectrometers" (Kevin Wong, MS), "Rapid Analyses of Carotenoids in Tomato Pastes Using Handheld Raman Spectroscopy" (Rachel Chen, MS) and "Development of a rapid and in-field phenotyping tool for screening protein quality in soybeans (Glycine max) using a miniature NIR sensor" (Xin Rong Sia). We established key collaborations with Thermo Nicolet, Rigaku, Si-Ware, Hamamatsu, Wasatch, Spectral Engines, Waters and Agilent to evaluate the applications of state-of-the-art portable handheld/portable spectrometers for real-time sensing of food. Since the 2016 to 2019 period of this project, over 20 undergraduate and graduate students have participated at international conferences receiving 12 awards for their oral and/or poster presentations. How have the results been disseminated to communities of interest?The results have been disseminated at the ACS, IFT, Pittcon and SCIX events that target scientific and industrial audiences interested in vibrational spectroscopy and food applications of the technology to monitor quality parameters and detect economic adulteration. In addition, I have presented our technology to various companies interested in deploying these handheld/portable devices for in-field data collection. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We have developed applications using vibrational spectroscopic techniques such as infrared (NIR and mid-IR) and Raman that can allow for reliable quality assurance and development of safety programs for high-risk ingredients. The proposed technology would enable detection and characterization of important quality traits and potential chemical food contaminants via unique spectral signature profiles, permitting real-time and field-based control of the product stream to enhance quality, safety and brand equity. We focused our efforts on the applications of palm-size NIR devices by establishing collaboration with Si-Ware Systems to implement their Micro Electro Mechanical Systems (MEMS) NIR sensor for food applications. The NIR device is developed with techniques inherited from the semiconductor microfabrication technologies providing extended NIR (1150 to 2500 nm) region compared to other NIR systems collecting spectra made up of overtones and combination bands that cover the full wavelength range giving the most accurate and reliable results. We evaluated the NIR performance in monitoring sugar levels in breakfast cereals, a popular choice especially for children due to its variety and ease, that besides providing important nutrients (whole grain or high in fiber) they contribute to sugar intake. Higher sugar intake by children has been shown through meta-analysis to be associated with a higher risk of obesity. The palm-size Neospectra unit provided excellent performances for determination of sucrose levels in breakfast cereals (SEP of 1.2% and R=0.99) with no sample preparation and fast results (20s) to allow the industry to monitor sugar content in commercial products to meet nutrition labeling requirements and allowing consumers to make informed diet decisions as growing concerns to risk of obesity from excessive sugar consumption. In addition, we have been testing the NIR device for rapid assessment of oleic acid in soybean breeding selections. By using soybean meal, we have developed models that can rapidly (15 sec) predict important quality traits in soybeans based on a unique spectral fingerprint. Novel NIR systems can provide the breeders and industry with a rapid method to determine oleic acid and other unique traits with equivalent levels of reliability and sensitivity as benchtop systems but allow for more flexibility since the unit can be easily be taken to the field. We worked with the tomato industry developing models interfaced to field-deployable portable infrared sensors for simultaneous prediction of multiple quality traits (pH, titratable acidity, °Brix, Bostwick viscosity, serum viscosity, fructose, glucose, vitamin C, citric acid and lycopene) in tomato paste, puree and sauce products with no sample preparation. We are completing the validation stage of the models and have included 3 years of materials with over 600 samples. The technology would permit the food industry for rapid (~1 min) screening for quality assurance applications that requires minimal sample preparation and personnel training and can be amenable for in-plant or in-field applications. In the field of authentication of raw ingredients, we have evaluated Raman instruments equipped with a 1064 nm laser to minimize the effect of fluorescence. The spectrum generated by a Raman instrument provided sharp and well-resolved bands, generating unique chemical information for developing robust untargeted approaches for identification of tampered samples. The capabilities of Raman have allowed to collect spectral data through clear glass/plastic containers to monitor their contents without perturbing the sample. Evaluation of a global set of honey samples showed the prevalence of adulteration in the market with 17% of store-bought samples labeled as "pure" were tainted with other ingredients. Our results are in accordance with reports from JRC Round Table and American Bee Keeping Association that ~ 14-23% of honeys in the market are adulterated. We have established collaboration with Universidad Nacional Agraria la Molina (UNALM) (Lima, Peru) for authentication of high-value Andean flours (Quinoa, amaranth and canihua). Andean grain flours exhibit many desired nutritional characteristics, such as high protein content, presence of essential amino acids, good source of dietary fiber, and being rich in the "heart healthy" Omega-3 and Omega-6 fatty acids. Due to the increased demand for these healthy grain alternatives there is risk of adulteration with less-expensive grains. Spectra was collected by using a portable infrared system with a single- reflection diamond ATR crystal, and analyzed by Soft Independent Model of Class Analogies and Partial Least Square Regression. Our data revealed a prevalence of adulteration in commercial samples from Peruvian markets. Regression models were generated for rapid determination of fat (Rcv = 0.98, SECV = 0.3) and protein (Rcv = 0.98, SECV = 0.96) levels. New generation of portable infrared devices provided a viable tool for chemical profiling allowing for rapid, "in-field", and reliable authentication of food ingredients Finally, we have evaluated the use of handheld NIR and FTIR spectrometers for screening acrylamide in frozen French Fries. The food industry is modifying processing operations to limit acrylamide formation as consumer concerns increase as a result of new scientific evidence or federal/state regulations. A limiting factor in evaluating manufacturing processes on acrylamide levels has been the cost of the analytical instrumentation such as tandem HPLC/MS/MS making it prohibiting for in-plant QC analysis. The industry is therefore in need for an alternative method that can provide data in a timely and economic manner so they can comply with the law and minimize costly recalls, loss of reputation, and costs associated with litigation. We have evaluated the feasibility of using vibrational spectroscopic techniques (NIR and Raman) to establish reliable monitoring program(s) for acrylamide levels. Overall, good linear correlation between the predicted acrylamide levels of independent sample set and actual measured acrylamide concentration by LC-MS/MS were found (rPred was ~0.90 and standard error of prediction (SEP) was 100 ppb). Our data supports the ability of vibrational spectroscopy for detection and quantification of acrylamide levels through spectral signature profiles enabling for high-throughput measurements for controlling the product stream and addressing risk management.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: ACS National Meeting & Expo. Division of Agricultural and Food Chemistry. AGFD 312. Testing the next generation of handheld devices for screening acrylamide in high-risk products. In Proposition 65 on Food Safety. Organizer: Michael Granvogl and Shaun Macmahon. Fall 2019. San Diego, CA, August 22nd  29th, 2019
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: SCIX 2019. FORENS-02 Food Forensics. Opportunities of Food Sensors for Field Applications: From Crop to Your Grocery Store. Co-chairs: Betsy Jean Yakes and Luis E. Rodriguez-Saona. Palm Springs, CA October 13th  18th, 2019.
• Type: Book Chapters Status: Published Year Published: 2018 Citation: Rodriguez-Saona, LE & Aykas, DP. 2018. Ch. 5: New Approaches for Rapid Tomato Quality Control. In "Tomato Chemistry, Industrial Processing and Product Development". Edited by Sebastiano Porretta. Royal Society of Chemistry.

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:The results from our Hatch project have been disseminated internationally and nationally through events that target scientific and industrial audiences interested in rapid alternatives for monitoring quality parameters and detection of food contaminants. These events provided a perfect setting for exchanging ideas with experts in the field, discuss current work, receive feedback from potential end-users, develop professional and industrial partnerships and disseminate knowledge through multidisciplinary collaborations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Our Lab has trained 2 OSU Food Science undergraduate students, Kuanrong Zhu and Wan Xin Tan. The students are working on authentication of maple syrup by Raman and sucrose level monitoring in breakfast cereals, respectively. Students are developing skills in spectral collection, chromatographic analysis and data analysis. We graduated 1 PhD and 1 MS students in Food Science at The Ohio State University with dissertation titles "The New Generation of Handheld Vibrational Spectroscopy Devices -Applications for Authentication of High Valued Commodities" and "Rapid Analyses of Carotenoids in Tomato Pastes Using Handheld Raman Spectroscopy", respectively. We continue key collaborations with Thermo Nicolet, Rigaku, Si-Ware, Hamamatsu, Wasatch and Agilent to evaluate the applications of state-of-the-art portable handheld/portable spectrometers for real-time sensing of food. Seven graduate students have presented at the 2018 IFT Annual Meeting and Food Expo. (009) In-Field Screening of Trans Fat Levels in Butters and Margarines Commercialized in the Peruvian Market Using a Handheld Infrared Spectrometer. Walter F. Salas Valerio, Luis E. Rodriguez-Saona, Fanny E. Ludena-Urquizo, Beatriz A. Hatta-Sakoda. Poster Session 3 - Tuesday, 7/17/2018, 11:00 am - 1:00 pm (038) In-Situ Monitoring of Biochemical Changes Occurring During the Ripening Process of Artisanal Turkish White Cheese Using Portable Mid-Infrared Spectrometer. HULYA YAMAN, Luis E. Rodriguez-Saona. Poster Session 1 - Dairy Foods Monday, 7/16/2018, 12:00 pm - 2:00 pm (073) Pomegranate Juice in the Spotlight: Rapid Authenticity Screening Based on Infrared (FTIR) Spectroscopy. Didem P. Aykas, Ahmed Menevseoglu, Luis E. Rodriguez-Saona. Poster Session 5 - Fruit & Vegetable Products - Wednesday, 7/18/2018, 9:00 am - 11:00 am (079) Rapid Analysis of Carotenoids in Fresh Tomatoes and Pastes Using Handheld Raman Spectroscopy. Yuju Chen, Luis E. Rodriguez-Saona. Poster Session 5 - Fruit & Vegetable Products - Wednesday, 7/18/2018, 9:00 am - 11:00 am (081) What's in Your Fruit Juice? Rapid Quality Screening Based on Infrared (FTIR) Spectroscopy. Didem P. Aykas, Luis E. Rodriguez-Saona. Poster Session 2 - Quality Assurance; Monday, 7/16/2018, 3:00 pm - 5:00 pm (085) Determination of Protected Designation of Origin (PDO) Peruvian Pisco Spirits by Portable FT-IR, Raman, and UV Spectroscopy and Multivariate Data Analysis. Ahmed Menevseoglu, Beatriz Hatta Sakoda, Juan C. Palma, Luis E. Rodriguez-Saona. Poster Session 2 - Quality Assurance- Monday, 7/16/2018, 3:00 pm - 5:00 pm (088) Authentication of Honeys Using Handheld Raman Spectroscopy. Mei-Ling Shotts, Columbus, OH, United States, Peren Aykas, Luis E. Rodriguez-Saona. Poster Session 2 - Quality Assurance - Monday, 7/16/2018, 3:00 pm - 5:00 pm How have the results been disseminated to communities of interest?The results have been disseminated at the ACS, IFT, Pittcon and SCIX events that target scientific and industrial audiences interested in vibrational spectroscopy and food applications of the technology to monitor quality parameters and detect economic adulteration. In addition, I have been invited to present our technology to various companies that have interest in deploying these handheld/portable devices for in-field data collection. What do you plan to do during the next reporting period to accomplish the goals?We will continue our efforts of monitoring quality traits in olive oil and other edible oils. We will work on developing rapid phenotypic tools for secreening spoilage bacteria in tomato paste products.We are interested in developing sensors fro detection of acrylamide in various food products and detection of pesticides. We will evaluate the capabilities of handheld NIR and Raman technology for detection of contaminants using SERS substrates for enhancing the Raman features and minimize fluorescence interference.

Impacts
What was accomplished under these goals? We continue to develop applications using vibrational spectroscopic techniques such as infrared (NIR and mid-IR) and Raman that can allow for reliable quality assurance and development of safety programs for high-risk ingredients. The proposed technology would enable detection and characterization of important quality traits and potential microbial and chemical food contaminants via unique spectral signature profiles, permitting real-time and field-based control of the product stream to enhance quality, safety and brand equity. This year we have focused our efforts on the applications of palm-size NIR devices. We have established collaboration with Si-Ware Systems to implement their Micro Electro Mechanical Systems (MEMS) NIR sensor for food applications. The NIR device is developed with techniques inherited from the semiconductor microfabrication technologies providing extended NIR (1150 to 2500 nm) region compared to other handheld NIR systems.Currently we are evaluating its performance in monitoring sugar levels in breakfast cereals, a popular choice especially for children due to its variety and ease, however, besides providing important nutrients (whole grain or high in fiber) they contribute to sugar intake. Higher sugar intake by children has been shown through meta-analysis to be associated with a higher risk of obesity. The palm-size Neospectra unit provided excellent performances for determination of sucrose levels in breakfast cereals (SEP of 1.2% and R=0.99) with no sample preparation and fast results (20s) to allow the industry to monitor sugar content in commercial products to meet nutrition labeling requirements and allowing consumers to make informed diet decisions as growing concerns to risk of obesity from excessive sugar consumption. In addition, we have been testing the NIR device for rapid assessment of oleic acid in soybean breeding selections. By using soybean meal, we have developed models that can rapidly (<1 min) predict important quality traits in soybeans based on a unique spectral fingerprint. Novel NIR systems may provide the breeders and industry with a rapid method to determine oleic acid and other unique traits with equivalent levels of reliability and sensitivity as benchtop systems but allow for more flexibility since the unit can be easily taken to the field. In the field of authentication of raw ingredients, we have been evaluating Raman instruments equipped with a 1060 nm laser to minimize the effect of fluorescence. The spectrum generated by a Raman instrument provides sharper and well-resolved bands, generating unique chemical information for developing robust untargeted approaches for identification of tampered samples. The capabilities of Raman have allowed to collect spectral data through clear glass/plastic containers to monitor their contents without perturbing the sample. Evaluation of a global set of honey samples showed the prevalence of adulteration in the market with 23% of store-bought samples labeled as "pure" were tainted with other ingredients. Our results are in accordance with reports from JRC Round Table and American Bee Keeping Association that approximately 14-23% of honeys in the market are adulterated. We continue to work with Universidad Nacional Agraria la Molina (UNALM) (Lima, Peru) for authentication of high-value Andean flours (Quinoa, amaranth and canihua). Andean grain flours exhibit many desired nutritional characteristics, such as high protein content, presence of essential amino acids, good source of dietary fiber, and being rich in the "heart healthy" Omega-3 and Omega-6 fatty acids. Due to the increased demand for these healthy grain alternatives there is risk of adulteration with less-expensive grains. Spectra was collected by using a portable infrared system with a single- reflection diamond ATR crystal, and analyzed by Soft Independent Model of Class Analogies and Partial Least Square Regression. Our data revealed a prevalence of adulteration in commercial samples from Peruvian markets. Regression models were generated for rapid determination of fat (Rcv = 0.98, SECV = 0.3) and protein (Rcv = 0.98, SECV = 0.96) levels. New generation of portable infrared devices provided a viable tool for chemical profiling allowing for rapid, "in-field", and reliable authentication of food ingredients Finally, we continue evaluating the use of handheld Raman and NIR spectrometers for screening acrylamide in frozen French Fries. The food industry is modifying processing operations to limit acrylamide formation as consumer concerns increase as a result of new scientific evidence or federal/state regulations. A limiting factor in evaluating manufacturing processes on acrylamide levels has been the cost of the analytical instrumentation such as tandem HPLC/MS/MS making it prohibiting for in-plant QC analysis. The industry is therefore in need for an alternative method that can provide data in a timely and economic manner so they can comply with the law and minimize costly recalls, loss of reputation, and costs associated with litigation. We have evaluated the feasibility of using vibrational spectroscopic techniques (NIR and Raman) to establish reliable monitoring program(s) for acrylamide levels. Overall, good linear correlation between the predicted acrylamide levels of independent sample set and actual measured acrylamide concentration by LC-MS/MS were found (rPred was ~0.90 and standard error of prediction (SEP) was 100 ppb). Our data supports the ability of vibrational spectroscopy for detection and quantification of acrylamide levels through spectral signature profiles enabling for high-throughput measurements for controlling the product stream and addressing risk management.

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: Santosh Dhakal, V.M. Balasubramaniam, Huseyin Ayvaz, and Luis Rodriguez-Saona. 2018. Kinetic modeling of ascorbic acid degradation of pineapple juice subjected to combined pressure-thermal treatment. Journal of Food Engineering 224, 62-70.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Dunham NT, Rodriguez-Saona LE. 2018 Nutrient intake and balancing among female Colobus angolensis palliatus inhabiting structurally distinct forest areas: Effects of group, season, and reproductive state. Am J Primatol. 80(7): e22878.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Shotts, M.-L., Plans Pujolras, M., Rossell, C., Rodriguez-Saona, L. 2018. Authentication of indigenous flours (Quinoa, Amaranth and ka�iwa) from the Andean region using a portable ATR-Infrared device in combination with pattern recognition analysis, Journal of Cereal Science. 82:65-72
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Caterina Villari, Arnaud Dowkiw, Rasmus Enderle, Marjan Ghasemkhani, Thomas Kirisits, Erik D. Kj�r, Diana Mar?iulynien?, Lea V. McKinney, Berthold Metzler, Facundo Mu�oz, Lene R. Nielsen, Alfas Pli?ra, Lars-G�ran Stener, Vytautas Suchockas, Luis Rodriguez-Saona, Pierluigi Bonello & Michelle Cleary. 2018. Advanced spectroscopy-based phenotyping offers a potential solution to the ash dieback epidemic. Scientific Reports 8:17448 | DOI:10.1038/s41598-018-35770-0.

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:The results from our Hatch project have been disseminated internationally and nationally through events that target scientific and industrial audiences interested in rapid alternatives for monitoring quality parameters and detection of food contaminants. These events provided a perfect setting for exchanging ideas with experts in the field, discuss current work, receive feedback from potential end-users, develop professional and industrial partnerships and disseminate knowledge through multidisciplinary collaborations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Our Lab has trained 3 OSU Food Science undergraduate students, Chris Huang, Anna Schmenck and Regina Tanuwidjaja. The students partcipated in several projects including spectral collection, chromatographic analysis and data analysis. Anna presented her project entitled "Non-targeted screening for GMO high-oleic soybeans and non-GMO conventional soybeans by vibrational spectroscopy" at the 2017 IFT Annual Metting and Expo that took place in Las Vegas in June 25th-28th and took third place in the undergraduate research competition. We graduated 1 MS students in Food Science at The Ohio State University working on detection of acrylamide in French fries. We continue key collaborations with Thermo Nicolet and Agilent to evaluate the applications of state-of-the-art portable handheld/portable spectrometers for real-time sensing of food. Five graduate students have presented at the 2017 IFT Annual Meeting and Food Expo. How have the results been disseminated to communities of interest?The results have been disseminated at the ACS, 9th International Workshop on Anthocyanins, IFT and SCIX events that target scientific and industrial audiences interested in vibrational spectroscopy and food applications of the technology to monitor quality parameters and detect economic adulteration. In additiona, I have been invited to present our technology to various companies that have interest in deploying these handheld/portable devices for in-field data collection. What do you plan to do during the next reporting period to accomplish the goals?We will continue our efforts of monitoring quality traits in tomato products, olive oil and other edible oils, detection of acrylamide in various food products and detection of pesticides. We will evaluate the capabilities of Raman technology for detection of contaminants using SERS substrates for enhancing the Raman features and minimize fluorescence interferance.

Impacts
What was accomplished under these goals? We continue to develop applications using vibrational spectroscopic techniques such as infrared (NIR and mid-IR) and Raman that can allow for reliable quality assurance and development of safety programs for high-risk ingredients. The proposed technology would enable detection and characterization of important quality traits and potential microbial and chemical food contaminants via unique spectral signature profiles, permitting real-time and field-based control of the product stream to enhance quality, safety and brand equity. This year we have focussed our efforts on the applications of handheld Raman instruments equipped with a 1060 nm laser to minimize the effect of fluorescence. The spectrum generated by a Raman instrument generally has sharper and better resolved peaks than NIR, providing unique chemical information for developing robust untargeted approaches for identification of unknown samples. In addition, the Raman excitation laser can be transmitted through clear glass containers and have been used to monitor their contents without perturbing the sample. We are exploring the use of Raman fingerprinting for authentication of honey samples collecting the spectra through glass containers. Our data shows that a handheld Raman device can identify adulterated honey samples without disturbing the containers. Furthermore, we are using the Raman spectra to identify plenish soybeans from conventional soybeans that have been developed to accumulate high oleic acid levels. We continue to work with the tomato industry developing models interfaced to field-deployable portable infrared sensors for simultaneous prediction of multiple quality traits (pH, titratable acidity, °Brix, Bostwick viscosity, serum viscosity, fructose, glucose, vitamin C, citric acid and lycopene) in tomato paste, puree and sauce products with no sample preparation. We are completing the validation stage of the models and have included 2 years of materials with over 600 samples. The technology would permit the food industry a rapid tool (~1 min) for screening and quality assurance applications that requires minimal sample preparation and personnel training and can be amenable for in-plant or in-field applications. In addition, we are completing a project in collaboration withUniversidad Nacional Agraria la Molina (UNALM) (Lima, Peru) for authentication of high-value Andean flours (Quinoa, amaranth and canihua). Andean grain flours exhibit many desired nutritional characteristics, such as high protein content, presence of essential amino acids, good source of dietary fiber, and being rich in the "heart healthy" Omega-3 and Omega-6 fatty acids. Due to the increased demand for these healthy grain alternatives there is risk of adulteration with less-expensive grains. Our results showed that there was some prevalence of adulteration in the peruvian market. Overall, a rapid analytical method was found that can be used "in-field" and provides the accurate recognition of adulterated food ingredients, making it a great alterative to conventional testing methods. We are also evaluating the use of handheld Raman and NIR spectrometers for screening acrylamide in frozen French Fries. The food industry is modifying processing operations to limit acrylamide formation as consumer concerns increase as a result of new scientific evidence or federal/state regulations. A limiting factor in evaluating manufacturing processes on acrylamide levels has been the cost of the analytical instrumentation such as tandem HPLC/MS/MS making it prohibiting for in-plant QC analysis. The industry is therefore in need for an alternative method that can provide data in a timely and economic manner so they can comply with the law and minimize costly recalls, loss of reputation, and costs associated with litigation. We have evaluated the feasibility of using vibrational spectroscopic techniques (NIR and Raman) to establish reliable monitoring program(s) for acrylamide levels. Overall, good linear correlation between the predicted acrylamide levels of independent sample set and actual measured acrylamide concentration by LC-MS/MS were found (rPred was ~0.90 and standard error of prediction (SEP) was 100 ppb). Our data supports the ability of vibrational spectroscopy for detection and quantification of acrylamide levels through spectral signature profiles enabling for high-throughput measurements for controlling the product stream and addressing risk management.

Publications

• Type: Book Chapters Status: Published Year Published: 2017 Citation: Rodriguez-Saona, LE & Aykas, DP. 2017. Ch. 9: Instrumentation. In Spectroscopic Methods in Food Analysis. Edited by Adriana S. Franca and Leo M.L. Nollet. CRC Press Taylor & Francis Group.
• Type: Books Status: Published Year Published: 2017 Citation: Rodriguez-Saona, L, Ayvaz, H, and Wehling RL. 2017. Ch. 8: Infrared and Raman Spectroscopy. In Food Analysis Fifth Edition. S. Suzanne Nielsen Editor. Springer Intl Publishing.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Infrared Spectroscopy: Effective Tool to Evaluate the Protein-Anthocyanin Complexation in Purple Corn. 9th International Workshop on Anthocyanins. Phytochemistry and Analysis session. 22nd - 24th February 2017, Auckland, New Zealand.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Portable Vibrational Spectroscopy Technologies for Screening Economically-Motivated Adulteration of Food Ingredients. IV Congreso Internacional Sobre Innovaci�n y Tendencias en Procesamiento de Alimentos y al XIX Congreso Nacional de Ciencia y Tecnolog�a de Alimentos. May 17  19, 2017.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Authentication of food ingredients by vibrational spectroscopy: Moving out of the lab. AGFD Award Symposium in honor of Dr. RE. Wrolstad. 254th ACS National Meeting & Exposition, August 20-24, 2017- Washington, DC.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Portable/Handheld infrared spectrometers becoming a reality for the Food Industry use of portable spectrometers in food industry processes. Advances in Applications of Handheld/Portable Spectrometers Session. SciX 2017, October 8-13, 2017 in Reno, NV.

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:The results from our project have been disseminated internationally and nationally through events that target scientific and industrial audiences interested in monitoring key quality parameters and detection of food contaminants. These events provided a perfect setting for exchanging ideas with experts in the field, discuss current work, receive feedback from the scientific community, develop new professional networks and disseminate knowledge through multidisciplinary research collaborations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We have trained 3 undergraduate students that supported research activities. We graduated 2 MS students in Food Science and Technology at The Ohio State University working on identification of E. coli serovars and rapid screening of quality traits in tomato paste. We continue key collaborations with Thermo Nicolet and Agilent to evaluate the applications of state-of-the-art portable handheld/portable spectrometers for real-time sensing of food. Two graduate students presented their work at the AOCS meeting in Salt Lake City (Utah). Three graduate students have presented at the 2016 IFT Annual Meeting and Food Expo at the McCormick Place, Chicago, IL. In addition, we demonstrated the capabilities of the infrared technology to 25 participants of 2016 Wilbur A. Gould Total Quality Management Short Course for the Snack Food Industry (Snack Food Association). How have the results been disseminated to communities of interest?The results have been disseminated at IFPAC, AOCS and IFT events that target scientific and industrial audiences interested in vibrational spectroscopy and food applications of the technology to monitor quality parameters and detect economic adulteration. What do you plan to do during the next reporting period to accomplish the goals?We will continue our efforts of monitoring quality traits in tomato products, edible oils, detection of trans fat and quantifying acrylamide in various food products. We will continue our efforts in Raman technology for detection of foodborne pathogens using SERS substrates for enhancing the Raman features.

Impacts
What was accomplished under these goals? We have negotiated to upgrade our state-of-the-art JASCO NRS-4100 Micro Raman instrument equipped with a 780nm and 560 nm lasers to the JASCO NRS-4100 with 1064nm laser that will provide unique capabilities for analysis of biological material because it minimizes fluorescence. We are also evaluating portable and handheld Raman instruments equipped with a 1060 nm laser. We continue working with the tomato industry validating models interfaced to field-deployable portable infrared sensors for simultaneous prediction of multiple quality traits (pH, titratable acidity, °Brix, Bostwick viscosity, serum viscosity, fructose, glucose, vitamin C, citric acid and lycopene) in tomato paste with no sample preparation. Our group continues our work establishing a reliable authentication program for extra virgin olive oils and edible oils and assessing quality parameters (i.e. peroxide value, free fatty acids, fatty acid profile, pyropheophytins and diglyceride levels) using portable mid-infrared spectroscopic techniques combined with pattern recognition analysis. The technology allows screening and quality assurance applications requiring minimal sample preparation, personnel training and can be amenable for in-plant or in-field applications. We are also developing predictive models to quantify trans-fat levels in bakery food products using portable mid-infrared (MIR) spectrometers coupled with a temperature control (65ºC). A total of 100 bakery and snack products produced using partially hydrogenated oils have been collected from local groceries and Internet purchases. Linear regression models using the negative second derivative at 966 cm-1 and PLSR multivariate models have shown excellent goodness of fit (R2 >0.9) and SEP of <0.5 % for a validation set of samples. Portable FTIR are ideal tools for regulatory applications and well suited to quality control applications spectroscopy, especially since the FDA has revoked the GRAS (generally recognized as safe) status of partially hydrogenated oils. In addition, we are generating authentication models for Andean flours (Quinoa, amaranth and canihua). Consumers in the United States have become more health conscious and have placed much more importance on a food's nutritional benefits. Andean grain flours exhibit many desired nutritional characteristics, such as high protein content, presence of essential amino acids, good source of dietary fiber, and being rich in the "heart healthy" Omega-3 and Omega-6 fatty acids. Due to the increased demand for these healthy grain alternatives there is risk of adulteration with less-expensive grains. In a collaboration with Universidad Nacional Agraria la Molina (UNALM) (Lima, Peru), we are completing the models for profiling Andean flours and identify potential economic adulteration, that is prevalent in the peruvian local market. Overall, a rapid analytical method was found that can be used "in-field" and provides the accurate recognition of adulterated food ingredients, making it a great alterative to conventional testing methods. We have established collaboration with Dr. Joshua Daniels from the Department of Veterinary Clinical Sciences at The Ohio State University to investigate E. coli identification by surface enhanced Raman spectroscopy. Our preliminary data shows the ability of SERS to discriminate among E.coli serovars.

Publications

• Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: M. Shotts, M. Plans-Pujolres, and L.E. Rodriguez-Samna. 2016. Application of Mid-infrared Portable Spectrometery in Determination of Trans-fatty Acid Content in Bakery Products. ANA 2: Green Analytical Techniques. AOCS Annual Conference. Salt Lake City, Utah.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Aykas Didem P., Mortas, Mustafa, and Rodriguez-Saona, Luis E. 2016. (017) Application of Portable Infrared Spectrometers With Pattern Recognition Analysis in Determination of Ground Coffee Quality. Session P02. Institute of Food Technologists Annual Meeting, Chicago, IL.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Zhang, Congcong, Aykas, Didem P. and Rodriguez-Saona, Luis E. 2016. (064) Rapid Assessment of Sugars and Organic Acids in Tomato Paste Using Portable Mid-Infrared Spectroscopy and Multivariate Analysis. Session P02. Institute of Food Technologists Annual Meeting, Chicago, IL.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Shouts, Mei-Ling, Rodriguez-Saona, Luis E. 2016. (099) Application of Mid-Infrared Portable Spectrometery in Determination of Trans-Fatty Acid Content in Bakery Products. Session P03. Institute of Food Technologists Annual Meeting, Chicago, IL.
• Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Aykas, DP and Rodriguez-Saona, LE. 2016. Assessing Potato Chip Oil Quality using a Portable Infrared Spectrometer Combined with Pattern Recognition Analysis. Anal Methods. 8: 731  741
• Type: Book Chapters Status: Accepted Year Published: 2016 Citation: Rodriguez-Saona, LE; Plans Pujolras, M; Giusti, MM. 2016. Targeted and non-targeted analysis. In: Analytical Separation Science, Vol 5: Sample preparation, method validation, and applications. J Anderson, A Berthod, V Pino, AM Stalcup (Ed). J Wiley & Sons, Inc. 1401-1436.

Progress 05/04/15 to 09/30/15

Outputs
Target Audience:Theresults from our project have been disseminated internationally and nationally through events that targetscientific and industrial audiences interested in monitoring key quality parameters anddetection of food contaminants. These eventsprovided a perfect setting for exchanging ideas with experts in the field, discuss current work, receive feedbackfrom the scientific community, develop new professional networks and disseminate knowledge through multidisciplinaryresearch collaborations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Our Lab has trained 2 Brazilian exchange students,Layla Dutra Brambila and Erika Borges Ferreira, partcipating in the Brazil Scientific Mobility Program (BSMP). The studentspartcipated in sevral projects including spectral collection in Andean flours, chromatographic analysis and data analysis. We have also trained 3 undergraduate students that supported research activities. We graduated1 MS studentsin Food Science atThe Ohio State University working on detection of acrylamide in French fries. We continue key collaborations with Thermo Nicoletand Agilentto evaluate the applications of state-of-the-art portable handheld/portable spectrometers for real-time sensing of food.Four graduate students have presented at the 2015 IFT Annual Meeting and Food Expo at the McCormick Place, Chicago, IL. In addition, we demonstrated the capabilities of the infrared technology to 30 partcipants of2015 Wilbur A. Gould Total Quality Management Short Course for the Snack Food Industry (Snack Food Association). How have the results been disseminated to communities of interest?The results have been disseminated at IFT and SCIX events that targetscientific and industrial audiences interested in vibrational spectroscopy and food applications of the technology to monitor quality parameters and detect economic adulteration. What do you plan to do during the next reporting period to accomplish the goals?We will continue our efforts of monitoring quality traits in tomato products, olive oil and other edible oils, detection of trans fat and quantifying acrylamide in various food products. We will evaluate the capabilities of Raman technology for detection of foodborne pathogens using SERS substrates for enhancing the Raman features and minimize fluorescence interferance.

Impacts
What was accomplished under these goals? Cutting edge sensor technologies can provide a valuable window into in-process food manufacturing to permit optimization of production rate, quality and safety of many food products. Vibrational spectroscopic techniques such as infrared (NIR and mid-IR) and Ramancan allow reliable quality assurance and safety programs for high-risk ingredients . The proposed technology would enable detection and characterization of important quality traits and potential microbial and chemical food contaminants via unique spectral signature profiles, permitting real-time and field-based control of the product stream to enhance quality, safety and brand equity. We have acquired a state-of-the-artJASCO NRS-4100 Micro Raman instrument equipped with a 780nm and 560 nm lasers and a permanent observation of the laser spot allowing for quick positioning and confidence that the area being measured is the area viewed. A separate laser/Raman scattering beamsplitter is used to direct the laser excitation into the microscope sampling optics and collect Raman scattering allowing for both maximum and optimized signal and scatter rejection. We are also evaluating portable and handheld Raman instruments equipped with a 1060 nm laser to minimize the effect of fluorescence. We are working with thetomato industry developing models interfaced to field-deployable portable infrared sensors for simultaneous prediction of multiple quality traits (pH, titratable acidity, °Brix, Bostwick viscosity, serum viscosity, fructose, glucose, vitamin C, citric acid and lycopene) in tomato paste, puree and sauce products with no sample preparation. Our group is alsoestablishing a reliable authentication program forextra virgin olive oils and edible oils and assessing quality parameters (i.e. peroxide value, free fatty acids, p-anisidine value, pyropheophytins) using portable mid-infrared spectroscopic techniques combined with pattern recognition analysis. The technology would allow detection of potential mislabeling and adulteration of edible oils providing spectral signature profiles that would permit the food industry a rapid tool (~1 min) for screening and quality assurance applications that requires minimal sample preparation and personnel training and can be amenable for in-plant or in-field applications. We are also developingpredictive models to quantify trans-fat levels in bakery food products using portable mid-infrared (MIR) spectrometers coupled with a temperature control (65ºC). Different approaches are tested using 30 standards gravimetrically prepared andextracted fat of different bakery and snack products (n=100). Linear regression models using the negative second derivative of the prepared standardsat 966 cm-1 andPLSR multivariate models showed a excellent goodness of fit (R2 >0.9)and SEP of 0.5 % for a validation set of samples. Determination of trans-fat using portable ATR-MIR spectrometers allowed to increase flexibility in set up and use while retaining the traditional benefits of FTIR spectroscopy, rapid throughput, high accuracy, and high amount of data per second, making it ideal for regulatory applications and well suited to quality control applications. In addition, we are generating authentication models for Andean flours (Quinoa, amaranth and canihua).consumers in the United States have become more health conscious and have placed much more importance on a food's nutritional benefits. Andean grain flours exhibit many desired nutritional characteristics, such as high protein content, presence of essential amino acids, good source of dietary fiber, and being rich in the "heart healthy" Omega-3 and Omega-6 fatty acids. Due to the increased demand for these healthy grain alternatives there is risk of adulteration with less-expensive grains. We are profilingAndean flours and ingredients provided by Universidad Nacional Agraria la Molina (UNALM) (Lima, Peru) and characterizing the fatty acids, aminoacids and protein levels of flours by using reference methods.Our results showedthat there was some prevalence of adulteration in the peruvian market. Overall, a rapid analytical method was found that can be used "in-field" and provides the accurate recognition of adulterated food ingredients, making it a great alterative to conventional testing methods.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: (365) Portable/Handheld Infrared Spectrometers becoming a Reality for the Food Industry; Luis Rodriguez-Saona; The Ohio State University. ADVANCES IN APPLICATIONS OF HANDHELD/PORTABLE SPECTROMETERS Organizer and Presider: Jason Rodriguez. SCIX 2015, Providence, RI.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ayvaz, H, Plans, M, Towers, BN, Auer, A, Rodriguez-Saona, LE. 2015. The use of infrared spectrometers to predict quality parameters of cornmeal (corn grits) and differentiate between organic and conventional practices. J Cereal Sci. 62: 22-30.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ayvaz H, Santos AM, Moyseenko J, Kleinhenz M, Rodriguez-Saona LE. 2015. Application of a portable infrared instrument for simultaneous analysis of sugars, asparagine and glutamine levels in raw potato tubers. Plant Foods Hum Nutr. 70(2):215-20
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Luis E. Rodriguez-Saona. Conferencia Internacional - Adulteraci�n en Alimentos. Facultad de Industrias Alimentarias, Universidad Nacional Agraria-La Molina. Agosto, 2015. Lima, Peru.