The Experts below are selected from a list of 2133 Experts worldwide ranked by ideXlab platform

Vittori Sauro - One of the best experts on this subject based on the ideXlab platform.

  • . Simultaneous quantification of 30 different bioactive compounds including polyphenols in spent coffee ground and coffee silverskin by HPLC-MS/MS triple quadrupole.
    Sociedade Portuguesa de Química, 2019
    Co-Authors: Caprioli Giovanni, Angeloni Simone, Kamgang Nzekoue, Astride Franks, Navarini Luciano, Sagratini Gianni, Vittori Sauro
    Abstract:

    Coffee is one of the most consumed beverages worldwide and according to the latest statistics the global consumption is more than 150 million of 60 kg coffee bags per year. As a consequence, large amount of coffee residues need to be disposed of [1]. Although many studies related to coffee and health have been carried out in green, ground and espresso coffee, two components of the coffee processing chain have been less investigated, i.e. coffee silverskin (CS) and spent coffee ground (SCG), the two main by-products presenting a huge potential to be valorized [2]. It is well known that coffee is a rich source of polyphenols, phytochemicals associated with many biological activities, and they are partially extracted from the beans during brewing; so, the resulting spent coffee is still a rich source of polyphenolic compounds. Similarly, silverskin is also considered to be a potential source of polyphenolic compounds because this tegument is in direct contact with the beans; therefore, it keeps part of the polyphenolic compounds constituents present in coffee beans [3]. On these bases, general aim of this project was to deepen the knowledge on CS and SCG, in the future perspective of their re-use as nutraceuticals by a) developing an efficient and low-cost method to extract different bioactive compounds including polyphenols from these by-products b) quantifying these bioactive compounds (chlorogenic acids, caffeine and other important coffee polyphenols such as flavonoids and phenolic acids) from different CS and SCG extracts by developing new HPLC-MS/MS analytical method. SCG and CS samples were extracted by using different solvents and extraction procedures (i.e. magnetic stirrer, ultrasound extraction). From an analytical point of view, UHPLC–MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology equipped with an ESI source operating in negative ionization mode. The separation was achieved using a Kinetex C18 analytical column (50 mm × 2.10 mm i.d., 2.6 μm) using a binary gradient of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The method was sensitive (LOQs for all compounds were in the range 1-100 μg kg-1), linear (R2 for all analytes was higher than 0.9907), accurate and robust. The best extraction procedure in term of amount of bioactive compounds extracted was ultrasound extraction with a mixture of ethanol:water 70:30 v/v; in this case, the highest amount of total bioactive compounds concentration was found. In the future, the most promising extracts in terms of bioactive compounds will be tested to evaluate theirs prebiotic and antimicrobial activity, in the perspective of their re-use as nutraceuticals

  • A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples
    Divisione di Spettrometria di massa e Università degli Studi di Camerino, 2019
    Co-Authors: Angeloni Simone, Caprioli Giovanni, Navarini Luciano, Sagratini Gianni, Khamitova Gulzhan, Vittori Sauro
    Abstract:

    A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples Simone Angeloni1,2, Giovanni Caprioli1, Gulzhan Khamitova1,2, Luciano Navarini3, Gianni Sagratini1, Sauro Vittori1 1 School of Pharmacy, University of Camerino, Camerino, Italy; 2 International Hub for Coffee Research and Innovation, Belforte del Chienti (MC), Italy; 3 illycaffè S.p.A., Trieste, Italy Summary: an analytical method for quantification of lignans (lariciresinol, matairesinol and secoisolariciresinol) and isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee has been developed by using HPLC-MS/MS. Results showed that the best extraction process was a base hydrolysis followed by enzymatic digestion and lignans were more abundant than isoflavones. Keywords: green coffee, phytoestrogen, HPLC-MS/MS 1 Introduction Coffee is one of the most important agricultural products in the international trade and last year 68 million 60 kg bags of green coffee were produced [1]. Green coffee beans, the starting raw material for roasted coffee and coffee beverages, are constituted by carbohydrates (55-65.5%), lipids (10-18%), nitrogen containing compounds (11-15%), purine alkaloids (0.8-4.0%), chlorogenic acids (6.7-9.2%) and minerals (3-5.4%). Other molecules that are found in lower percentages are non-volatile aliphatic acids (citric, malic and quinic acids) and phenols such as phytoestrogens [2]. The most studied and best-known phytoestrogens in foodstuffs are isoflavones and lignans. Both classes have been investigated in coffee powder and beverages but to the best of our knowledge, none have quantified them in green coffee. Hence, we sought to develop a simply and fast method to quantify three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee beans by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). For that purpose, firstly, we evaluated different extraction processes and the best one was chosen for lignan and isoflavone extraction. Secondly, we set up an efficient and fast HPLC-MS/MS method for simultaneous quantification of target molecules in green coffee. Finally, after validation the selected method was applied to 25 green coffee samples. 2 Experimental HPLC-MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology (Santa Clara, CA) equipped with an electrospray ionization (ESI) source operating in negative and positive ionization mode. The separation of target compounds was achieved on a Kinetex C18 analytical column (50 mm x 2.10 mm i.d., 2.6 μm) from Phenomenex (Castel Maggiore, Bologna, Italy). The mobile phase for HPLC-MS/MS analyses was a mixture of 85% water (A) and 15% HPLC-grade acetonitrile (B), both with 0.1% formic acid. The separation was obtained by flowing at 0.4 mL/min with gradient elution. Detection was performed in the “multiple reaction monitoring” (MRM) mode. Different extraction processes such as acid and base hydrolysis, enzymatic digestions, organic solvent extraction and a combination of these, were tested and recovery and quantitative data were examined as well. The best one was chosen and applied to 24 Coffea arabica samples having different geographical origins and 1 Coffeea canephora. Chemical differences among the various coffee samples, in terms of lignan and isoflavone contents, were analysed by PCA using the statistical software STATISTICA v.7.1 (Stat Soft Italia S.r.l., Vigonza, Italy). 3 Results A new analytical method for simultaneous quantitation of three lignans and six isoflavones was developed by using HPLC-MS/MS triple quadrupole. For isoflavones in glycosidic form, the precursor ions were protonated molecules [M+H]+ in positive polarity, whereas for the aglyconic form and lignans the precursor ions were deprotonated molecules [M-H]- in negative polarity. The method showed good linearity (R2 for all target compounds were equal to or higher than 0.9952), sensitivity (LODs for isoflavones and lignans ranged from 0.1 to 15 µg L-1) and the separation of studied molecules was obtained within 6 min. The best performing process was a double extraction composed of base hydrolysis in methanol and enzymatic digestion with clara-diastase, since it showed good recovery levels, ranging from 74 to 94%, and the highest total concentration of all the compounds studied (1193.4 μg kg-1). Therefore, this process was chosen and applied to 25 green coffee samples. Results showed that lignans (286.5-8131.8 μg kg-1) were more abundant than isoflavones (3.4-300.0 μg kg-1) and secoisolariciresinol (172.6-5714.1 μg kg-1) and lariciresinol (113.9-2417.7 μg kg-1) were the most abundant compounds followed by genistin (12.6-204.8 μg kg-1). After PCA analysis we found that Ethiopian samples differed significantly from the other origins for the higher content of secoisolariciresinol and lariciresinol suggesting that they are a good source of lignans. 4 Conclusions For the first time, a new analytical method for the simultaneous quantitation in green coffee of three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) has been developed, validated and then, applied to 25 green coffee samples. This work provided new knowledge into two important phytonutrients (isoflavones and lignans) in green coffee, one of the most important agricultural products in the international trade. References 1. International Coffee Organization (ICO). Statistics. Trade Statistic Tables. (2018). http://www.ico.org/prices/po-production.pdf. Accessed 25 August 2019. 2. S. I. Mussatto, E. M. Machado, S. Martins, J. A. Teixeira; Food and Bioprocess Technology, 4 (2011), pp 661-672

  • Quantification of Riboflavin, Nicotinamide and Nicotinic Acid in Anchovies (Engraulis Enrasicolus) by HPLC-MS/MS Triple Quadrupole
    Divisione di spettrometria di massa Società Chimica italiana, 2017
    Co-Authors: Caprioli Giovanni, Sagratini Gianni, Vittori Sauro, Torregiani Elisabetta
    Abstract:

    Simultaneous quantification of riboflavin, nicotinamide and nicotinic acid in anchovies (Engraulis enrasicolus) by HPLC-MS/MS triple quadrupole Giovanni Caprioli1, Sauro Vittori1, Gianni Sagratini1, Elisabetta Torregiani1 1School of Pharmacy, University of Camerino, Camerino, Italy Summary: A new analytical method that uses high performance liquid chromatography–triple quadrupole mass spectrometry (HPLC-MS/MS) was developed for the simultaneous analysis of vitamins B2 (riboflavin) and B3 (nicotinamide and nicotinic acid) in anchovies. Keywords: riboflavin, niacin, anchovies 1 Introduction Vitamins are crucial for maintaining good health in humans; lack of a sufficient amount of them can cause serious disease [1]. Essential for the calculation of dietary intake of nutrients from food is the use of reliable, accurate and precise analytical methods for nutrients in the foods. The water-soluble vitamins act mainly as coenzymes, while the fat-soluble ones act in different and more complex ways [2]. In foodstuffs, the vitamins B2 (riboflavin) and B3 (niacin) may be present in free (riboflavin, nicotinamide and nicotinic acid) and binding forms (essentially riboflavin-5’-phosphate (FMN), riboflavin- 5’-adenosyldiphosphate (FAD), nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP)). Furthermore, they may be bound tightly but non-covalently to proteins and polysaccharides [3]. Anchovies are a good source of those two vitamins, based on the various nutrition tables available. Thus, the aim of this paper was to evaluate the content of vitamin B2 (riboflavin) and B3 (niacin, i.e. nicotinic acid plus nicotinamide) in anchovies samples provided by a company called “Dimar Sapore di Mare”. 2 Experimental Anchovies canned in olive or sunflower oil were purchased from the local supermarket, while normal anchovy samples were kindly supplied by D.I.MAR. Srl - Sapore di Mare (Corridonia, Macerata, Italy). Different extraction procedures were tested and the best one was chosen for the accurate quantification of riboflavin, nicotinic acid and nicotinamide in different types of anchovy samples. Extraction Method n°1 consisted in acidic hydrolysis: ground anchovies were hydrolysed with 30 mL of 0.1 N HC1 at 100 °C for 30 min under magnetic stirring. Method n°2 consisted in the addition of TCA 50% for protein precipitation after acidic hydrolysis. Method n°3 consisted in acidic hydrolysis plus enzymatic hydrolysis with different enzymes or mixture of them (claradiastase, papain, takadiastase and amylase). Method n°4 consisted only in the enzymatic digestion. UHPLC–MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology (Santa Clara, CA) equipped with an ESI source operating in positive ionization mode. The separation of riboflavin, nicotinic acid and nicotinamide was achieved using a Kinetex Hilic analytical column (100 mm × 4.6 mm i.d., particle size 2.6 µm) from Phenomenex (Torrance, CA, USA). Detection was performed by electrospray ionization (ESI)-MS in the “multiple reaction monitoring” (MRM) mode. The MRM peaks areas were integrated for quantification; the most abundant product ion was used for quantification, and the rest of the products ions were used for qualification. 3 Results The analytical procedure developed is fast (2.5 minutes of chromatography run time), the method is sensitive (LOQ for all compounds is in the range 1-5 µg kg-1), linear (R2 for all compounds was higher than 0.9996), accurate and robust as it is possible to apply the method to both normal and under-oil/canned anchovies. The results obtained indicated that method 1 was the best of the four optimized extraction procedures in terms of recoveries (range 96.2 - 105.3%, with RSDs lower than 9.2%), quantitative results obtained, and rapidity, and thus it was chosen for the analysis of vitamins B2 and B3 in 23 anchovy samples (both normal and canned ones). The application to real samples indicates that the highest niacin levels were found in normal anchovies (101.21-150.92 mg kg-1), while the highest level of riboflavin was in anchovies canned in olive oil (ranging from 2.90 to 3.35 mg kg-1). 4 Conclusions A new analytical method that uses HPLC-MS/MS triple quadrupole was developed for the first time for the analysis of riboflavin, nicotinamide and nicotinic acid in anchovies. Samples were extracted using different extraction procedures, and acidic hydrolysis (method 1) was found to be the best in terms of recoveries, quantitative results and rapidity. In order to understand the dietary intake of vitamins from different types of anchovies, the method was applied to 23 samples including eleven normal anchovy samples and twelve canned ones. References 1. A. Zafra-Gomez, A. Garballo, J.C. Morales, L.E. Garcia-Ayuso. Journal of Agricultural and Food Chemistry, 54 (2006), pp. 4531-4536. 2. A. Gentili, F. Caretti, G. D’Ascenzo, S. Marchese, D. Perret, D. Di Corcia, L.M. Rocca. Rapid Communications in Mass Spectrometry, 22 (2008), pp. 2029-2043. 3. S. Ndaw, M. Bergaentzlè, D. Aoudè-Werner, C. Hasselmann. Enzymatic extraction procedure for the liquid chromatographic determination of niacin in foodstuffs. Food Chemistry, 78 (2002), pp. 129-134

Caprioli Giovanni - One of the best experts on this subject based on the ideXlab platform.

  • . Simultaneous quantification of 30 different bioactive compounds including polyphenols in spent coffee ground and coffee silverskin by HPLC-MS/MS triple quadrupole.
    Sociedade Portuguesa de Química, 2019
    Co-Authors: Caprioli Giovanni, Angeloni Simone, Kamgang Nzekoue, Astride Franks, Navarini Luciano, Sagratini Gianni, Vittori Sauro
    Abstract:

    Coffee is one of the most consumed beverages worldwide and according to the latest statistics the global consumption is more than 150 million of 60 kg coffee bags per year. As a consequence, large amount of coffee residues need to be disposed of [1]. Although many studies related to coffee and health have been carried out in green, ground and espresso coffee, two components of the coffee processing chain have been less investigated, i.e. coffee silverskin (CS) and spent coffee ground (SCG), the two main by-products presenting a huge potential to be valorized [2]. It is well known that coffee is a rich source of polyphenols, phytochemicals associated with many biological activities, and they are partially extracted from the beans during brewing; so, the resulting spent coffee is still a rich source of polyphenolic compounds. Similarly, silverskin is also considered to be a potential source of polyphenolic compounds because this tegument is in direct contact with the beans; therefore, it keeps part of the polyphenolic compounds constituents present in coffee beans [3]. On these bases, general aim of this project was to deepen the knowledge on CS and SCG, in the future perspective of their re-use as nutraceuticals by a) developing an efficient and low-cost method to extract different bioactive compounds including polyphenols from these by-products b) quantifying these bioactive compounds (chlorogenic acids, caffeine and other important coffee polyphenols such as flavonoids and phenolic acids) from different CS and SCG extracts by developing new HPLC-MS/MS analytical method. SCG and CS samples were extracted by using different solvents and extraction procedures (i.e. magnetic stirrer, ultrasound extraction). From an analytical point of view, UHPLC–MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology equipped with an ESI source operating in negative ionization mode. The separation was achieved using a Kinetex C18 analytical column (50 mm × 2.10 mm i.d., 2.6 μm) using a binary gradient of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The method was sensitive (LOQs for all compounds were in the range 1-100 μg kg-1), linear (R2 for all analytes was higher than 0.9907), accurate and robust. The best extraction procedure in term of amount of bioactive compounds extracted was ultrasound extraction with a mixture of ethanol:water 70:30 v/v; in this case, the highest amount of total bioactive compounds concentration was found. In the future, the most promising extracts in terms of bioactive compounds will be tested to evaluate theirs prebiotic and antimicrobial activity, in the perspective of their re-use as nutraceuticals

  • A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples
    Divisione di Spettrometria di massa e Università degli Studi di Camerino, 2019
    Co-Authors: Angeloni Simone, Caprioli Giovanni, Navarini Luciano, Sagratini Gianni, Khamitova Gulzhan, Vittori Sauro
    Abstract:

    A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples Simone Angeloni1,2, Giovanni Caprioli1, Gulzhan Khamitova1,2, Luciano Navarini3, Gianni Sagratini1, Sauro Vittori1 1 School of Pharmacy, University of Camerino, Camerino, Italy; 2 International Hub for Coffee Research and Innovation, Belforte del Chienti (MC), Italy; 3 illycaffè S.p.A., Trieste, Italy Summary: an analytical method for quantification of lignans (lariciresinol, matairesinol and secoisolariciresinol) and isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee has been developed by using HPLC-MS/MS. Results showed that the best extraction process was a base hydrolysis followed by enzymatic digestion and lignans were more abundant than isoflavones. Keywords: green coffee, phytoestrogen, HPLC-MS/MS 1 Introduction Coffee is one of the most important agricultural products in the international trade and last year 68 million 60 kg bags of green coffee were produced [1]. Green coffee beans, the starting raw material for roasted coffee and coffee beverages, are constituted by carbohydrates (55-65.5%), lipids (10-18%), nitrogen containing compounds (11-15%), purine alkaloids (0.8-4.0%), chlorogenic acids (6.7-9.2%) and minerals (3-5.4%). Other molecules that are found in lower percentages are non-volatile aliphatic acids (citric, malic and quinic acids) and phenols such as phytoestrogens [2]. The most studied and best-known phytoestrogens in foodstuffs are isoflavones and lignans. Both classes have been investigated in coffee powder and beverages but to the best of our knowledge, none have quantified them in green coffee. Hence, we sought to develop a simply and fast method to quantify three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee beans by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). For that purpose, firstly, we evaluated different extraction processes and the best one was chosen for lignan and isoflavone extraction. Secondly, we set up an efficient and fast HPLC-MS/MS method for simultaneous quantification of target molecules in green coffee. Finally, after validation the selected method was applied to 25 green coffee samples. 2 Experimental HPLC-MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology (Santa Clara, CA) equipped with an electrospray ionization (ESI) source operating in negative and positive ionization mode. The separation of target compounds was achieved on a Kinetex C18 analytical column (50 mm x 2.10 mm i.d., 2.6 μm) from Phenomenex (Castel Maggiore, Bologna, Italy). The mobile phase for HPLC-MS/MS analyses was a mixture of 85% water (A) and 15% HPLC-grade acetonitrile (B), both with 0.1% formic acid. The separation was obtained by flowing at 0.4 mL/min with gradient elution. Detection was performed in the “multiple reaction monitoring” (MRM) mode. Different extraction processes such as acid and base hydrolysis, enzymatic digestions, organic solvent extraction and a combination of these, were tested and recovery and quantitative data were examined as well. The best one was chosen and applied to 24 Coffea arabica samples having different geographical origins and 1 Coffeea canephora. Chemical differences among the various coffee samples, in terms of lignan and isoflavone contents, were analysed by PCA using the statistical software STATISTICA v.7.1 (Stat Soft Italia S.r.l., Vigonza, Italy). 3 Results A new analytical method for simultaneous quantitation of three lignans and six isoflavones was developed by using HPLC-MS/MS triple quadrupole. For isoflavones in glycosidic form, the precursor ions were protonated molecules [M+H]+ in positive polarity, whereas for the aglyconic form and lignans the precursor ions were deprotonated molecules [M-H]- in negative polarity. The method showed good linearity (R2 for all target compounds were equal to or higher than 0.9952), sensitivity (LODs for isoflavones and lignans ranged from 0.1 to 15 µg L-1) and the separation of studied molecules was obtained within 6 min. The best performing process was a double extraction composed of base hydrolysis in methanol and enzymatic digestion with clara-diastase, since it showed good recovery levels, ranging from 74 to 94%, and the highest total concentration of all the compounds studied (1193.4 μg kg-1). Therefore, this process was chosen and applied to 25 green coffee samples. Results showed that lignans (286.5-8131.8 μg kg-1) were more abundant than isoflavones (3.4-300.0 μg kg-1) and secoisolariciresinol (172.6-5714.1 μg kg-1) and lariciresinol (113.9-2417.7 μg kg-1) were the most abundant compounds followed by genistin (12.6-204.8 μg kg-1). After PCA analysis we found that Ethiopian samples differed significantly from the other origins for the higher content of secoisolariciresinol and lariciresinol suggesting that they are a good source of lignans. 4 Conclusions For the first time, a new analytical method for the simultaneous quantitation in green coffee of three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) has been developed, validated and then, applied to 25 green coffee samples. This work provided new knowledge into two important phytonutrients (isoflavones and lignans) in green coffee, one of the most important agricultural products in the international trade. References 1. International Coffee Organization (ICO). Statistics. Trade Statistic Tables. (2018). http://www.ico.org/prices/po-production.pdf. Accessed 25 August 2019. 2. S. I. Mussatto, E. M. Machado, S. Martins, J. A. Teixeira; Food and Bioprocess Technology, 4 (2011), pp 661-672

  • Quantification of Riboflavin, Nicotinamide and Nicotinic Acid in Anchovies (Engraulis Enrasicolus) by HPLC-MS/MS Triple Quadrupole
    Divisione di spettrometria di massa Società Chimica italiana, 2017
    Co-Authors: Caprioli Giovanni, Sagratini Gianni, Vittori Sauro, Torregiani Elisabetta
    Abstract:

    Simultaneous quantification of riboflavin, nicotinamide and nicotinic acid in anchovies (Engraulis enrasicolus) by HPLC-MS/MS triple quadrupole Giovanni Caprioli1, Sauro Vittori1, Gianni Sagratini1, Elisabetta Torregiani1 1School of Pharmacy, University of Camerino, Camerino, Italy Summary: A new analytical method that uses high performance liquid chromatography–triple quadrupole mass spectrometry (HPLC-MS/MS) was developed for the simultaneous analysis of vitamins B2 (riboflavin) and B3 (nicotinamide and nicotinic acid) in anchovies. Keywords: riboflavin, niacin, anchovies 1 Introduction Vitamins are crucial for maintaining good health in humans; lack of a sufficient amount of them can cause serious disease [1]. Essential for the calculation of dietary intake of nutrients from food is the use of reliable, accurate and precise analytical methods for nutrients in the foods. The water-soluble vitamins act mainly as coenzymes, while the fat-soluble ones act in different and more complex ways [2]. In foodstuffs, the vitamins B2 (riboflavin) and B3 (niacin) may be present in free (riboflavin, nicotinamide and nicotinic acid) and binding forms (essentially riboflavin-5’-phosphate (FMN), riboflavin- 5’-adenosyldiphosphate (FAD), nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP)). Furthermore, they may be bound tightly but non-covalently to proteins and polysaccharides [3]. Anchovies are a good source of those two vitamins, based on the various nutrition tables available. Thus, the aim of this paper was to evaluate the content of vitamin B2 (riboflavin) and B3 (niacin, i.e. nicotinic acid plus nicotinamide) in anchovies samples provided by a company called “Dimar Sapore di Mare”. 2 Experimental Anchovies canned in olive or sunflower oil were purchased from the local supermarket, while normal anchovy samples were kindly supplied by D.I.MAR. Srl - Sapore di Mare (Corridonia, Macerata, Italy). Different extraction procedures were tested and the best one was chosen for the accurate quantification of riboflavin, nicotinic acid and nicotinamide in different types of anchovy samples. Extraction Method n°1 consisted in acidic hydrolysis: ground anchovies were hydrolysed with 30 mL of 0.1 N HC1 at 100 °C for 30 min under magnetic stirring. Method n°2 consisted in the addition of TCA 50% for protein precipitation after acidic hydrolysis. Method n°3 consisted in acidic hydrolysis plus enzymatic hydrolysis with different enzymes or mixture of them (claradiastase, papain, takadiastase and amylase). Method n°4 consisted only in the enzymatic digestion. UHPLC–MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology (Santa Clara, CA) equipped with an ESI source operating in positive ionization mode. The separation of riboflavin, nicotinic acid and nicotinamide was achieved using a Kinetex Hilic analytical column (100 mm × 4.6 mm i.d., particle size 2.6 µm) from Phenomenex (Torrance, CA, USA). Detection was performed by electrospray ionization (ESI)-MS in the “multiple reaction monitoring” (MRM) mode. The MRM peaks areas were integrated for quantification; the most abundant product ion was used for quantification, and the rest of the products ions were used for qualification. 3 Results The analytical procedure developed is fast (2.5 minutes of chromatography run time), the method is sensitive (LOQ for all compounds is in the range 1-5 µg kg-1), linear (R2 for all compounds was higher than 0.9996), accurate and robust as it is possible to apply the method to both normal and under-oil/canned anchovies. The results obtained indicated that method 1 was the best of the four optimized extraction procedures in terms of recoveries (range 96.2 - 105.3%, with RSDs lower than 9.2%), quantitative results obtained, and rapidity, and thus it was chosen for the analysis of vitamins B2 and B3 in 23 anchovy samples (both normal and canned ones). The application to real samples indicates that the highest niacin levels were found in normal anchovies (101.21-150.92 mg kg-1), while the highest level of riboflavin was in anchovies canned in olive oil (ranging from 2.90 to 3.35 mg kg-1). 4 Conclusions A new analytical method that uses HPLC-MS/MS triple quadrupole was developed for the first time for the analysis of riboflavin, nicotinamide and nicotinic acid in anchovies. Samples were extracted using different extraction procedures, and acidic hydrolysis (method 1) was found to be the best in terms of recoveries, quantitative results and rapidity. In order to understand the dietary intake of vitamins from different types of anchovies, the method was applied to 23 samples including eleven normal anchovy samples and twelve canned ones. References 1. A. Zafra-Gomez, A. Garballo, J.C. Morales, L.E. Garcia-Ayuso. Journal of Agricultural and Food Chemistry, 54 (2006), pp. 4531-4536. 2. A. Gentili, F. Caretti, G. D’Ascenzo, S. Marchese, D. Perret, D. Di Corcia, L.M. Rocca. Rapid Communications in Mass Spectrometry, 22 (2008), pp. 2029-2043. 3. S. Ndaw, M. Bergaentzlè, D. Aoudè-Werner, C. Hasselmann. Enzymatic extraction procedure for the liquid chromatographic determination of niacin in foodstuffs. Food Chemistry, 78 (2002), pp. 129-134

Daniele Valdes - One of the best experts on this subject based on the ideXlab platform.

  • rapid spe lc ms ms analysis for atrazine its by products simazine and s metolachlor in groundwater samples
    MethodsX, 2020
    Co-Authors: Helene Blanchoud, Fabrice Alliot, Ningxin Chen, Daniele Valdes
    Abstract:

    Abstract A rapid analysis of pesticides using on-line Solid phase extraction LC MS/MS (Agilent Technology) was performed using only 2-mL water samples. SPE cartridge PLRP-s was used for the pre-concentration sample with methanol elution in back flush. Sensitive transitions and mass spectrometry conditions were optimized by direct infusion of individual standard solutions in a positive electrospray mode. Water samples were spiked with internal standards to compensate the matrix effect. The limit of quantification was calculated to be 20 ng L−1 using the standard deviation of blank analysis injected ten times and uncertainties were estimated at less than 20% on concentrations. This method was validated to study leaching water samples for which only small quantities of water were available. • Only 2 mL water sample was used. • Samples were filtered at 0.2 µm and spiked with individual standard. • Compounds were separated in an 18.5-min elution time using the dynamic MRM program.

Sagratini Gianni - One of the best experts on this subject based on the ideXlab platform.

  • . Simultaneous quantification of 30 different bioactive compounds including polyphenols in spent coffee ground and coffee silverskin by HPLC-MS/MS triple quadrupole.
    Sociedade Portuguesa de Química, 2019
    Co-Authors: Caprioli Giovanni, Angeloni Simone, Kamgang Nzekoue, Astride Franks, Navarini Luciano, Sagratini Gianni, Vittori Sauro
    Abstract:

    Coffee is one of the most consumed beverages worldwide and according to the latest statistics the global consumption is more than 150 million of 60 kg coffee bags per year. As a consequence, large amount of coffee residues need to be disposed of [1]. Although many studies related to coffee and health have been carried out in green, ground and espresso coffee, two components of the coffee processing chain have been less investigated, i.e. coffee silverskin (CS) and spent coffee ground (SCG), the two main by-products presenting a huge potential to be valorized [2]. It is well known that coffee is a rich source of polyphenols, phytochemicals associated with many biological activities, and they are partially extracted from the beans during brewing; so, the resulting spent coffee is still a rich source of polyphenolic compounds. Similarly, silverskin is also considered to be a potential source of polyphenolic compounds because this tegument is in direct contact with the beans; therefore, it keeps part of the polyphenolic compounds constituents present in coffee beans [3]. On these bases, general aim of this project was to deepen the knowledge on CS and SCG, in the future perspective of their re-use as nutraceuticals by a) developing an efficient and low-cost method to extract different bioactive compounds including polyphenols from these by-products b) quantifying these bioactive compounds (chlorogenic acids, caffeine and other important coffee polyphenols such as flavonoids and phenolic acids) from different CS and SCG extracts by developing new HPLC-MS/MS analytical method. SCG and CS samples were extracted by using different solvents and extraction procedures (i.e. magnetic stirrer, ultrasound extraction). From an analytical point of view, UHPLC–MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology equipped with an ESI source operating in negative ionization mode. The separation was achieved using a Kinetex C18 analytical column (50 mm × 2.10 mm i.d., 2.6 μm) using a binary gradient of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The method was sensitive (LOQs for all compounds were in the range 1-100 μg kg-1), linear (R2 for all analytes was higher than 0.9907), accurate and robust. The best extraction procedure in term of amount of bioactive compounds extracted was ultrasound extraction with a mixture of ethanol:water 70:30 v/v; in this case, the highest amount of total bioactive compounds concentration was found. In the future, the most promising extracts in terms of bioactive compounds will be tested to evaluate theirs prebiotic and antimicrobial activity, in the perspective of their re-use as nutraceuticals

  • A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples
    Divisione di Spettrometria di massa e Università degli Studi di Camerino, 2019
    Co-Authors: Angeloni Simone, Caprioli Giovanni, Navarini Luciano, Sagratini Gianni, Khamitova Gulzhan, Vittori Sauro
    Abstract:

    A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples Simone Angeloni1,2, Giovanni Caprioli1, Gulzhan Khamitova1,2, Luciano Navarini3, Gianni Sagratini1, Sauro Vittori1 1 School of Pharmacy, University of Camerino, Camerino, Italy; 2 International Hub for Coffee Research and Innovation, Belforte del Chienti (MC), Italy; 3 illycaffè S.p.A., Trieste, Italy Summary: an analytical method for quantification of lignans (lariciresinol, matairesinol and secoisolariciresinol) and isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee has been developed by using HPLC-MS/MS. Results showed that the best extraction process was a base hydrolysis followed by enzymatic digestion and lignans were more abundant than isoflavones. Keywords: green coffee, phytoestrogen, HPLC-MS/MS 1 Introduction Coffee is one of the most important agricultural products in the international trade and last year 68 million 60 kg bags of green coffee were produced [1]. Green coffee beans, the starting raw material for roasted coffee and coffee beverages, are constituted by carbohydrates (55-65.5%), lipids (10-18%), nitrogen containing compounds (11-15%), purine alkaloids (0.8-4.0%), chlorogenic acids (6.7-9.2%) and minerals (3-5.4%). Other molecules that are found in lower percentages are non-volatile aliphatic acids (citric, malic and quinic acids) and phenols such as phytoestrogens [2]. The most studied and best-known phytoestrogens in foodstuffs are isoflavones and lignans. Both classes have been investigated in coffee powder and beverages but to the best of our knowledge, none have quantified them in green coffee. Hence, we sought to develop a simply and fast method to quantify three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee beans by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). For that purpose, firstly, we evaluated different extraction processes and the best one was chosen for lignan and isoflavone extraction. Secondly, we set up an efficient and fast HPLC-MS/MS method for simultaneous quantification of target molecules in green coffee. Finally, after validation the selected method was applied to 25 green coffee samples. 2 Experimental HPLC-MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology (Santa Clara, CA) equipped with an electrospray ionization (ESI) source operating in negative and positive ionization mode. The separation of target compounds was achieved on a Kinetex C18 analytical column (50 mm x 2.10 mm i.d., 2.6 μm) from Phenomenex (Castel Maggiore, Bologna, Italy). The mobile phase for HPLC-MS/MS analyses was a mixture of 85% water (A) and 15% HPLC-grade acetonitrile (B), both with 0.1% formic acid. The separation was obtained by flowing at 0.4 mL/min with gradient elution. Detection was performed in the “multiple reaction monitoring” (MRM) mode. Different extraction processes such as acid and base hydrolysis, enzymatic digestions, organic solvent extraction and a combination of these, were tested and recovery and quantitative data were examined as well. The best one was chosen and applied to 24 Coffea arabica samples having different geographical origins and 1 Coffeea canephora. Chemical differences among the various coffee samples, in terms of lignan and isoflavone contents, were analysed by PCA using the statistical software STATISTICA v.7.1 (Stat Soft Italia S.r.l., Vigonza, Italy). 3 Results A new analytical method for simultaneous quantitation of three lignans and six isoflavones was developed by using HPLC-MS/MS triple quadrupole. For isoflavones in glycosidic form, the precursor ions were protonated molecules [M+H]+ in positive polarity, whereas for the aglyconic form and lignans the precursor ions were deprotonated molecules [M-H]- in negative polarity. The method showed good linearity (R2 for all target compounds were equal to or higher than 0.9952), sensitivity (LODs for isoflavones and lignans ranged from 0.1 to 15 µg L-1) and the separation of studied molecules was obtained within 6 min. The best performing process was a double extraction composed of base hydrolysis in methanol and enzymatic digestion with clara-diastase, since it showed good recovery levels, ranging from 74 to 94%, and the highest total concentration of all the compounds studied (1193.4 μg kg-1). Therefore, this process was chosen and applied to 25 green coffee samples. Results showed that lignans (286.5-8131.8 μg kg-1) were more abundant than isoflavones (3.4-300.0 μg kg-1) and secoisolariciresinol (172.6-5714.1 μg kg-1) and lariciresinol (113.9-2417.7 μg kg-1) were the most abundant compounds followed by genistin (12.6-204.8 μg kg-1). After PCA analysis we found that Ethiopian samples differed significantly from the other origins for the higher content of secoisolariciresinol and lariciresinol suggesting that they are a good source of lignans. 4 Conclusions For the first time, a new analytical method for the simultaneous quantitation in green coffee of three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) has been developed, validated and then, applied to 25 green coffee samples. This work provided new knowledge into two important phytonutrients (isoflavones and lignans) in green coffee, one of the most important agricultural products in the international trade. References 1. International Coffee Organization (ICO). Statistics. Trade Statistic Tables. (2018). http://www.ico.org/prices/po-production.pdf. Accessed 25 August 2019. 2. S. I. Mussatto, E. M. Machado, S. Martins, J. A. Teixeira; Food and Bioprocess Technology, 4 (2011), pp 661-672

  • Quantification of Riboflavin, Nicotinamide and Nicotinic Acid in Anchovies (Engraulis Enrasicolus) by HPLC-MS/MS Triple Quadrupole
    Divisione di spettrometria di massa Società Chimica italiana, 2017
    Co-Authors: Caprioli Giovanni, Sagratini Gianni, Vittori Sauro, Torregiani Elisabetta
    Abstract:

    Simultaneous quantification of riboflavin, nicotinamide and nicotinic acid in anchovies (Engraulis enrasicolus) by HPLC-MS/MS triple quadrupole Giovanni Caprioli1, Sauro Vittori1, Gianni Sagratini1, Elisabetta Torregiani1 1School of Pharmacy, University of Camerino, Camerino, Italy Summary: A new analytical method that uses high performance liquid chromatography–triple quadrupole mass spectrometry (HPLC-MS/MS) was developed for the simultaneous analysis of vitamins B2 (riboflavin) and B3 (nicotinamide and nicotinic acid) in anchovies. Keywords: riboflavin, niacin, anchovies 1 Introduction Vitamins are crucial for maintaining good health in humans; lack of a sufficient amount of them can cause serious disease [1]. Essential for the calculation of dietary intake of nutrients from food is the use of reliable, accurate and precise analytical methods for nutrients in the foods. The water-soluble vitamins act mainly as coenzymes, while the fat-soluble ones act in different and more complex ways [2]. In foodstuffs, the vitamins B2 (riboflavin) and B3 (niacin) may be present in free (riboflavin, nicotinamide and nicotinic acid) and binding forms (essentially riboflavin-5’-phosphate (FMN), riboflavin- 5’-adenosyldiphosphate (FAD), nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP)). Furthermore, they may be bound tightly but non-covalently to proteins and polysaccharides [3]. Anchovies are a good source of those two vitamins, based on the various nutrition tables available. Thus, the aim of this paper was to evaluate the content of vitamin B2 (riboflavin) and B3 (niacin, i.e. nicotinic acid plus nicotinamide) in anchovies samples provided by a company called “Dimar Sapore di Mare”. 2 Experimental Anchovies canned in olive or sunflower oil were purchased from the local supermarket, while normal anchovy samples were kindly supplied by D.I.MAR. Srl - Sapore di Mare (Corridonia, Macerata, Italy). Different extraction procedures were tested and the best one was chosen for the accurate quantification of riboflavin, nicotinic acid and nicotinamide in different types of anchovy samples. Extraction Method n°1 consisted in acidic hydrolysis: ground anchovies were hydrolysed with 30 mL of 0.1 N HC1 at 100 °C for 30 min under magnetic stirring. Method n°2 consisted in the addition of TCA 50% for protein precipitation after acidic hydrolysis. Method n°3 consisted in acidic hydrolysis plus enzymatic hydrolysis with different enzymes or mixture of them (claradiastase, papain, takadiastase and amylase). Method n°4 consisted only in the enzymatic digestion. UHPLC–MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology (Santa Clara, CA) equipped with an ESI source operating in positive ionization mode. The separation of riboflavin, nicotinic acid and nicotinamide was achieved using a Kinetex Hilic analytical column (100 mm × 4.6 mm i.d., particle size 2.6 µm) from Phenomenex (Torrance, CA, USA). Detection was performed by electrospray ionization (ESI)-MS in the “multiple reaction monitoring” (MRM) mode. The MRM peaks areas were integrated for quantification; the most abundant product ion was used for quantification, and the rest of the products ions were used for qualification. 3 Results The analytical procedure developed is fast (2.5 minutes of chromatography run time), the method is sensitive (LOQ for all compounds is in the range 1-5 µg kg-1), linear (R2 for all compounds was higher than 0.9996), accurate and robust as it is possible to apply the method to both normal and under-oil/canned anchovies. The results obtained indicated that method 1 was the best of the four optimized extraction procedures in terms of recoveries (range 96.2 - 105.3%, with RSDs lower than 9.2%), quantitative results obtained, and rapidity, and thus it was chosen for the analysis of vitamins B2 and B3 in 23 anchovy samples (both normal and canned ones). The application to real samples indicates that the highest niacin levels were found in normal anchovies (101.21-150.92 mg kg-1), while the highest level of riboflavin was in anchovies canned in olive oil (ranging from 2.90 to 3.35 mg kg-1). 4 Conclusions A new analytical method that uses HPLC-MS/MS triple quadrupole was developed for the first time for the analysis of riboflavin, nicotinamide and nicotinic acid in anchovies. Samples were extracted using different extraction procedures, and acidic hydrolysis (method 1) was found to be the best in terms of recoveries, quantitative results and rapidity. In order to understand the dietary intake of vitamins from different types of anchovies, the method was applied to 23 samples including eleven normal anchovy samples and twelve canned ones. References 1. A. Zafra-Gomez, A. Garballo, J.C. Morales, L.E. Garcia-Ayuso. Journal of Agricultural and Food Chemistry, 54 (2006), pp. 4531-4536. 2. A. Gentili, F. Caretti, G. D’Ascenzo, S. Marchese, D. Perret, D. Di Corcia, L.M. Rocca. Rapid Communications in Mass Spectrometry, 22 (2008), pp. 2029-2043. 3. S. Ndaw, M. Bergaentzlè, D. Aoudè-Werner, C. Hasselmann. Enzymatic extraction procedure for the liquid chromatographic determination of niacin in foodstuffs. Food Chemistry, 78 (2002), pp. 129-134

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  • . Simultaneous quantification of 30 different bioactive compounds including polyphenols in spent coffee ground and coffee silverskin by HPLC-MS/MS triple quadrupole.
    Sociedade Portuguesa de Química, 2019
    Co-Authors: Caprioli Giovanni, Angeloni Simone, Kamgang Nzekoue, Astride Franks, Navarini Luciano, Sagratini Gianni, Vittori Sauro
    Abstract:

    Coffee is one of the most consumed beverages worldwide and according to the latest statistics the global consumption is more than 150 million of 60 kg coffee bags per year. As a consequence, large amount of coffee residues need to be disposed of [1]. Although many studies related to coffee and health have been carried out in green, ground and espresso coffee, two components of the coffee processing chain have been less investigated, i.e. coffee silverskin (CS) and spent coffee ground (SCG), the two main by-products presenting a huge potential to be valorized [2]. It is well known that coffee is a rich source of polyphenols, phytochemicals associated with many biological activities, and they are partially extracted from the beans during brewing; so, the resulting spent coffee is still a rich source of polyphenolic compounds. Similarly, silverskin is also considered to be a potential source of polyphenolic compounds because this tegument is in direct contact with the beans; therefore, it keeps part of the polyphenolic compounds constituents present in coffee beans [3]. On these bases, general aim of this project was to deepen the knowledge on CS and SCG, in the future perspective of their re-use as nutraceuticals by a) developing an efficient and low-cost method to extract different bioactive compounds including polyphenols from these by-products b) quantifying these bioactive compounds (chlorogenic acids, caffeine and other important coffee polyphenols such as flavonoids and phenolic acids) from different CS and SCG extracts by developing new HPLC-MS/MS analytical method. SCG and CS samples were extracted by using different solvents and extraction procedures (i.e. magnetic stirrer, ultrasound extraction). From an analytical point of view, UHPLC–MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology equipped with an ESI source operating in negative ionization mode. The separation was achieved using a Kinetex C18 analytical column (50 mm × 2.10 mm i.d., 2.6 μm) using a binary gradient of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The method was sensitive (LOQs for all compounds were in the range 1-100 μg kg-1), linear (R2 for all analytes was higher than 0.9907), accurate and robust. The best extraction procedure in term of amount of bioactive compounds extracted was ultrasound extraction with a mixture of ethanol:water 70:30 v/v; in this case, the highest amount of total bioactive compounds concentration was found. In the future, the most promising extracts in terms of bioactive compounds will be tested to evaluate theirs prebiotic and antimicrobial activity, in the perspective of their re-use as nutraceuticals

  • A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples
    Divisione di Spettrometria di massa e Università degli Studi di Camerino, 2019
    Co-Authors: Angeloni Simone, Caprioli Giovanni, Navarini Luciano, Sagratini Gianni, Khamitova Gulzhan, Vittori Sauro
    Abstract:

    A new HPLC-MS/MS analytical method for isoflavone and lignan quantification in 25 green coffee samples Simone Angeloni1,2, Giovanni Caprioli1, Gulzhan Khamitova1,2, Luciano Navarini3, Gianni Sagratini1, Sauro Vittori1 1 School of Pharmacy, University of Camerino, Camerino, Italy; 2 International Hub for Coffee Research and Innovation, Belforte del Chienti (MC), Italy; 3 illycaffè S.p.A., Trieste, Italy Summary: an analytical method for quantification of lignans (lariciresinol, matairesinol and secoisolariciresinol) and isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee has been developed by using HPLC-MS/MS. Results showed that the best extraction process was a base hydrolysis followed by enzymatic digestion and lignans were more abundant than isoflavones. Keywords: green coffee, phytoestrogen, HPLC-MS/MS 1 Introduction Coffee is one of the most important agricultural products in the international trade and last year 68 million 60 kg bags of green coffee were produced [1]. Green coffee beans, the starting raw material for roasted coffee and coffee beverages, are constituted by carbohydrates (55-65.5%), lipids (10-18%), nitrogen containing compounds (11-15%), purine alkaloids (0.8-4.0%), chlorogenic acids (6.7-9.2%) and minerals (3-5.4%). Other molecules that are found in lower percentages are non-volatile aliphatic acids (citric, malic and quinic acids) and phenols such as phytoestrogens [2]. The most studied and best-known phytoestrogens in foodstuffs are isoflavones and lignans. Both classes have been investigated in coffee powder and beverages but to the best of our knowledge, none have quantified them in green coffee. Hence, we sought to develop a simply and fast method to quantify three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) in green coffee beans by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). For that purpose, firstly, we evaluated different extraction processes and the best one was chosen for lignan and isoflavone extraction. Secondly, we set up an efficient and fast HPLC-MS/MS method for simultaneous quantification of target molecules in green coffee. Finally, after validation the selected method was applied to 25 green coffee samples. 2 Experimental HPLC-MS/MS studies were performed using an Agilent 1290 Infinity series and a Triple Quadrupole 6420 from Agilent Technology (Santa Clara, CA) equipped with an electrospray ionization (ESI) source operating in negative and positive ionization mode. The separation of target compounds was achieved on a Kinetex C18 analytical column (50 mm x 2.10 mm i.d., 2.6 μm) from Phenomenex (Castel Maggiore, Bologna, Italy). The mobile phase for HPLC-MS/MS analyses was a mixture of 85% water (A) and 15% HPLC-grade acetonitrile (B), both with 0.1% formic acid. The separation was obtained by flowing at 0.4 mL/min with gradient elution. Detection was performed in the “multiple reaction monitoring” (MRM) mode. Different extraction processes such as acid and base hydrolysis, enzymatic digestions, organic solvent extraction and a combination of these, were tested and recovery and quantitative data were examined as well. The best one was chosen and applied to 24 Coffea arabica samples having different geographical origins and 1 Coffeea canephora. Chemical differences among the various coffee samples, in terms of lignan and isoflavone contents, were analysed by PCA using the statistical software STATISTICA v.7.1 (Stat Soft Italia S.r.l., Vigonza, Italy). 3 Results A new analytical method for simultaneous quantitation of three lignans and six isoflavones was developed by using HPLC-MS/MS triple quadrupole. For isoflavones in glycosidic form, the precursor ions were protonated molecules [M+H]+ in positive polarity, whereas for the aglyconic form and lignans the precursor ions were deprotonated molecules [M-H]- in negative polarity. The method showed good linearity (R2 for all target compounds were equal to or higher than 0.9952), sensitivity (LODs for isoflavones and lignans ranged from 0.1 to 15 µg L-1) and the separation of studied molecules was obtained within 6 min. The best performing process was a double extraction composed of base hydrolysis in methanol and enzymatic digestion with clara-diastase, since it showed good recovery levels, ranging from 74 to 94%, and the highest total concentration of all the compounds studied (1193.4 μg kg-1). Therefore, this process was chosen and applied to 25 green coffee samples. Results showed that lignans (286.5-8131.8 μg kg-1) were more abundant than isoflavones (3.4-300.0 μg kg-1) and secoisolariciresinol (172.6-5714.1 μg kg-1) and lariciresinol (113.9-2417.7 μg kg-1) were the most abundant compounds followed by genistin (12.6-204.8 μg kg-1). After PCA analysis we found that Ethiopian samples differed significantly from the other origins for the higher content of secoisolariciresinol and lariciresinol suggesting that they are a good source of lignans. 4 Conclusions For the first time, a new analytical method for the simultaneous quantitation in green coffee of three lignans (lariciresinol, matairesinol and secoisolariciresinol) and six isoflavones (biochanin A, daidzein, daidzin, genistein, genistin and formononetin) has been developed, validated and then, applied to 25 green coffee samples. This work provided new knowledge into two important phytonutrients (isoflavones and lignans) in green coffee, one of the most important agricultural products in the international trade. References 1. International Coffee Organization (ICO). Statistics. Trade Statistic Tables. (2018). http://www.ico.org/prices/po-production.pdf. Accessed 25 August 2019. 2. S. I. Mussatto, E. M. Machado, S. Martins, J. A. Teixeira; Food and Bioprocess Technology, 4 (2011), pp 661-672