METABOLOMICS APPROACH FOR AUTHENTICATION OF PISCO AND DETECTION OF CONTAMINANTS Dissertation Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Ahmed Menevseoglu, M.Sc. Graduate Program in Food Science and Technology The Ohio State University 2019 Dissertation Committee Luis E. Rodriguez-Saona, Advisor M. Monica Giusti Christopher T. Simons Matthias S. Klein Copyrighted by Ahmed Menevseoglu 2019 2 Abstract Pisco is one of the most consumed distilled alcoholic drink in South America produced in Peru. The production and consumption of Pisco has been increasing in recent years. As a result of this, Pisco is becoming more prone to adulteration with a concerning of 32% of all alcoholic beverages in Peru is adulterated. Pisco obtains Protected Denomination of Origin (PDO) with very strict regulations. For instance, fermentation must be completely done, neither second distillation nor dilution with water is allowed. Rapid and reliable techniques are needed to authenticate the Pisco and to detect possible adulterations. The objective of our first study was to authenticate Pisco grape types based on non- destructive methods UV-Vis, and vibrational spectroscopy. Pisco obtains a Protected Denomination of Origin and based on that only certain grape types which can only be produced in certain areas of Peru can be used in Pisco manufacturing. A total of 136 Pisco samples (67 Quebranta, 44 Italia, 25 Torontel) were analyzed. Our data indicated that 34% of all analyzed Pisco were suspected to be adulterated with another type of grape. Benchtop and portable UV- Vis spectrometers were shown to be a very good tool to discriminate grape types into two groups aromatic and non-aromatic. FTIR showed a good distinction with two groups, however, pre- treatment of the samples was time consuming making UV-Vis would be more efficient tool for authentication of Pisco. The objective of our second study was to evaluate handheld Raman and portable FT-IR spectroscopy devices for non-invasive and rapid determination of methanol and ethanol levels in ii Peruvian Pisco. A total of 171 commercial Peruvian Pisco samples were analyzed. Raman spectrometers equipped either with a 1064nm (Rigaku Progeny) or 785nm (Metrohm Mira) excitation laser and a portable infrared spectroscopy (Agilent Cary 630) unit operating in absorption mode were used to predict ethanol and methanol levels in Pisco. GC-MS data revealed that 17% of Pisco samples had ethanol levels below 38% indicating possible water dilution. The levels of methanol ranged from 0.13 to 0.54 g/L, well below maximum levels for fruit brandies established by countries and international organizations to ensure the safety and quality of spirits. Handheld Raman equipped with a 1064nm excitation laser gave best results for determination of ethanol (SEP=1.4%; Rval=0.96) and methanol (SEP=0.04g/L; Rval=0.87). Portable/handheld vibrational spectroscopy combined with chemometrics allowed rapid (~1 min), non-invasive and in-situ screening of counterfeited brandies (Pisco) by dilution or substituting with cheaper alcohols. The objective of our third study was to determine chemical and metal contaminants in Pisco, and their predictions with portable UV-Vis, and mid-IR units. Ethyl carbamate and furfural by GC-MS, and metals (Cu, Al, Zn, Pb) by ICP-OES levels in Pisco were determined. Concentrations of EC, and furfural ranged from 0 – 256 ppb, 0.07 – 3.40 mg/100ml, respectively. Interestingly, 81 out of 148 analyzed Pisco samples showed higher levels of Cu (>5mg/l). Pb levels in four samples, and Zn levels in six Pisco samples were detected as higher than the allowed concentrations (Zn<1.5, Pb<0.5 mg/L). Benchtop and portable UV-Vis units allowed rapid determination of furfural (Rval>0.97) and titratable acidity (Rval>0.90) with very good prediction. Mid-IR unit showed very good correlation with titratable acidity (Rval>90), and iii copper levels (Rval>0.96). Handheld/portable UV-Vis and IR units can provide reliable and rapid assessments for quality control purposes of Pisco with less sample preparation and personnel training. Portable vibrational, and UV-Vis spectrometers can be used in real-time assessment for authentication purposes of fruit-based distilled alcoholic beverages. The industry may profit by using this technology because its simplicity, reliability, compatibility with traditional techniques, and cost efficiency. Keywords: Pisco, vibrational spectroscopy, authentication, adulteration, contamination iv Dedication Dedicated to my all family. v Acknowledgments I would like to thank Dr. Rodriguez-Saona (aka Lucho) for everything he did for me. He was more than an advisor to me, sometimes a good friend, sometimes a brother. I learnt a lot from him about how an academician should be, and I hope I can follow his path. My great appreciation for Dr. Giusti, she helped me a lot with personal and academic issues. It was good to know someone was there when you really needed. My committee members; Dr. Klein and Dr. Simons was very helpful during my PhD studies. They were so kind to me all the time. I would like to thank Beatriz Hatta-Sakoda and Sandra R.M. Garcia-Duclos from University of La Molina in Peru (UNALM), and Mr. Victor Toledo-Herrera (Pisco 1615 Inc.), providing the Pisco samples. I also would like to thank Ministry of National Education in Turkey that supported me financially for my PhD studies. My son Kursad Erdem, he deserves a big thank from me because I was not able to take care of him much during my PhD studies. I appreciated my wife, Betul, for her patience. She was always there stood by me through the good times and bad. My dad, my mom… I really thank them for all the things they have done for me. vi Vita October 19, 1987…………. Born – Sivas, Turkey 2005 – 2009………………. B.S. Food Engineering, Afyon Kocatepe University, Turkey 2011 – 2012………………. M.S. Food Engineering, Ankara University, Turkey 2015 – Present…………… GRA, Food Science and Technology, The Ohio State University Publications Menevseoglu A, Hatta-Sakoda B, Palma JC, Rodriguez-Saona LE. 2018. Determination of Protected Designation of Origin (PDO) Peruvian Pisco spirits by portable FT-IR, Raman, and UV spectroscopy and multivariate data analysis. Poster Presentation, IFT18, 15 - 18 July 2018, Chicago, IL, USA Fields of Study Major Field: Food Science and Technology vii Table of Contents Abstract ........................................................................................................................................... ii Dedication ........................................................................................................................................v Acknowledgments.......................................................................................................................... vi Vita ................................................................................................................................................ vii List of Tables ................................................................................................................................. xi List of Figures ............................................................................................................................... xii Chapter 1. Introduction ....................................................................................................................1 Chapter 2. Literature Review ...........................................................................................................4 2.1 Adulteration in Foods ................................................................................................................4 2.2 Adulteration in alcoholic beverages...........................................................................................6 2.2.1 Tequila and Mezcal ........................................................................................................ 8 2.2.2 Whisky ......................................................................................................................... 10 2.2.3 Vodka ........................................................................................................................... 11 2.2.4 Rum .............................................................................................................................. 12 2.2.5 Brazilian Cachaca ........................................................................................................ 12 2.2.6 Pisco ............................................................................................................................. 13 2.3 Adulteration types in alcoholic beverage and case studies ......................................................16 2.4 Detection methods for adulteration and authentication ...........................................................17 2.5 Evaluation of vibrational spectroscopy performance on authentication and adulteration of alcoholic beverages ........................................................................................................................18 2.6 References ................................................................................................................................19 Chapter 3. Untargeted approaches for characterization of Pisco based on the grape types ...........25 3.1 Abstract ....................................................................................................................................25