Introduction: Isoflavones and lignans are considered phytoestrogens, a group of secondary plant metabolites that possess some estrogenic and antiestrogenic activities [1]. The aim of our work was to develop a new analytical method for the extraction and quantification of three lignans and six isoflavones in green coffee by using HPLC-MS/MS triple quadrupole. Methods: Several extraction procedures were evaluated, including acid, basic and enzymatic hydrolysis and methanolic extractions. Two different enzymes, i.e. taka-diastase and clara-diastase, were tested and after a series of attempts, we tried to optimize the procedure combining acid or basic hydrolysis with enzymatic digestion. As analytical instrument, we chose an HPLC-MS/MS equipped with an ESI source. Results: The most performing extraction procedure was the alkaline hydrolysis in methanol plus enzymatic hydrolysis with clara-Diastase (the recoveries were from 74 to 94%). Then, our analytical method was validated using the following criteria: linearity (R2>0.995 for all compounds), inter-day and intra-day repeatability (RSD % in the range of 0.6-9.3 for inter-day repeatability and 0.4-4.2 for intra-day repeatability) and sensitivity (LODs in the range of 0.1-5 µg L-1, LOQs in the range of 1-16 µg L-1). Our analytical method was fast (the separation of the nine target compounds happened in less than 7 minutes) and avoided long sample preparation and purification procedures. Conclusions: For the first time, a new analytical method for the simultaneous quantification of isoflavones and lignans in green coffee beans has been developed. This method will be applied to various green coffee samples in order to prove the presence or the lack of these molecules in raw material of coffee. References 1. Kuhnle, G. G., Dell’Aquila, C., Aspinall, S. M., Runswick, S. A., Mulligan, A. A., Bingham, S. A., Journal of Agricultural and Food Chemistry, 56, 7311-7315, 2008.
A new extraction method to quantify lignans and isoflavones in green coffee using HPLC-MS/MS triple quadrupole.
Simone Angeloni;Giovanni Caprioli;Gianni Sagratini;Sauro Vittori.
2018-01-01
Abstract
Introduction: Isoflavones and lignans are considered phytoestrogens, a group of secondary plant metabolites that possess some estrogenic and antiestrogenic activities [1]. The aim of our work was to develop a new analytical method for the extraction and quantification of three lignans and six isoflavones in green coffee by using HPLC-MS/MS triple quadrupole. Methods: Several extraction procedures were evaluated, including acid, basic and enzymatic hydrolysis and methanolic extractions. Two different enzymes, i.e. taka-diastase and clara-diastase, were tested and after a series of attempts, we tried to optimize the procedure combining acid or basic hydrolysis with enzymatic digestion. As analytical instrument, we chose an HPLC-MS/MS equipped with an ESI source. Results: The most performing extraction procedure was the alkaline hydrolysis in methanol plus enzymatic hydrolysis with clara-Diastase (the recoveries were from 74 to 94%). Then, our analytical method was validated using the following criteria: linearity (R2>0.995 for all compounds), inter-day and intra-day repeatability (RSD % in the range of 0.6-9.3 for inter-day repeatability and 0.4-4.2 for intra-day repeatability) and sensitivity (LODs in the range of 0.1-5 µg L-1, LOQs in the range of 1-16 µg L-1). Our analytical method was fast (the separation of the nine target compounds happened in less than 7 minutes) and avoided long sample preparation and purification procedures. Conclusions: For the first time, a new analytical method for the simultaneous quantification of isoflavones and lignans in green coffee beans has been developed. This method will be applied to various green coffee samples in order to prove the presence or the lack of these molecules in raw material of coffee. References 1. Kuhnle, G. G., Dell’Aquila, C., Aspinall, S. M., Runswick, S. A., Mulligan, A. A., Bingham, S. A., Journal of Agricultural and Food Chemistry, 56, 7311-7315, 2008.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
