INFOGEST inter-laboratory recommendations for assaying gastric and pancreatic lipases activities prior to in vitro digestion studies
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INFOGEST inter-laboratory recommendations for assaying gastric and pancreatic lipases activities prior to in vitro digestion studies. / Grundy, Myriam M.L.; Abrahamse, Evan; Almgren, Annette; Alminger, Marie; Andres, Ana; Ariëns, Renata M.C.; Bastiaan-Net, Shanna; Bourlieu-Lacanal, Claire; Brodkorb, André; Bronze, Maria R.; Comi, Irene; Couëdelo, Leslie; Dupont, Didier; Durand, Annie; El, Sedef N.; Grauwet, Tara; Heerup, Christine; Heredia, Ana; Infantes Garcia, Marcos R.; Jungnickel, Christian; Kłosowska-Chomiczewska, Ilona E.; Létisse, Marion; Macierzanka, Adam; Mackie, Alan R.; McClements, David J.; Menard, Olivia; Meynier, Anne; Michalski, Marie Caroline; Mulet-Cabero, Ana Isabel; Mullertz, Anette; Payeras Perelló, Francina M.; Peinado, Irene; Robert, Mélina; Secouard, Sébastien; Serra, Ana T.; Silva, Sandra D.; Thomassen, Gabriel; Tullberg, Cecilia; Undeland, Ingrid; Vaysse, Carole; Vegarud, Gerd E.; Verkempinck, Sarah H.E.; Viau, Michelle; Zahir, Mostafa; Zhang, Ruojie; Carrière, Frédéric.
I: Journal of Functional Foods, Bind 82, 104497, 2021.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - INFOGEST inter-laboratory recommendations for assaying gastric and pancreatic lipases activities prior to in vitro digestion studies
AU - Grundy, Myriam M.L.
AU - Abrahamse, Evan
AU - Almgren, Annette
AU - Alminger, Marie
AU - Andres, Ana
AU - Ariëns, Renata M.C.
AU - Bastiaan-Net, Shanna
AU - Bourlieu-Lacanal, Claire
AU - Brodkorb, André
AU - Bronze, Maria R.
AU - Comi, Irene
AU - Couëdelo, Leslie
AU - Dupont, Didier
AU - Durand, Annie
AU - El, Sedef N.
AU - Grauwet, Tara
AU - Heerup, Christine
AU - Heredia, Ana
AU - Infantes Garcia, Marcos R.
AU - Jungnickel, Christian
AU - Kłosowska-Chomiczewska, Ilona E.
AU - Létisse, Marion
AU - Macierzanka, Adam
AU - Mackie, Alan R.
AU - McClements, David J.
AU - Menard, Olivia
AU - Meynier, Anne
AU - Michalski, Marie Caroline
AU - Mulet-Cabero, Ana Isabel
AU - Mullertz, Anette
AU - Payeras Perelló, Francina M.
AU - Peinado, Irene
AU - Robert, Mélina
AU - Secouard, Sébastien
AU - Serra, Ana T.
AU - Silva, Sandra D.
AU - Thomassen, Gabriel
AU - Tullberg, Cecilia
AU - Undeland, Ingrid
AU - Vaysse, Carole
AU - Vegarud, Gerd E.
AU - Verkempinck, Sarah H.E.
AU - Viau, Michelle
AU - Zahir, Mostafa
AU - Zhang, Ruojie
AU - Carrière, Frédéric
N1 - Publisher Copyright: © 2021
PY - 2021
Y1 - 2021
N2 - In vitro digestion studies often use animal digestive enzyme extracts as substitutes of human gastric and pancreatic secretions. Pancreatin from porcine origin is thus commonly used to provide relevant pancreatic enzymes such as proteases, amylase and lipase. Rabbit gastric extracts (RGE) have been recently introduced to provide gastric lipase in addition to pepsin. Before preparing simulated gastric and pancreatic extracts with targeted enzyme activities as described in in vitro digestion protocols, it is important to determine the activities of enzyme preparations using validated methods. The purpose of this inter-laboratory study within the INFOGEST network was to test the repeatability and reproducibility of lipase assays using the pH-stat technique for measuring the activities of gastric and pancreatic lipases from various sources. Twenty-one laboratories having different pH-stat devices received the same protocol with identical batches of RGE and two pancreatin sources. Lipase assays were performed using tributyrin as a substrate and three different amounts (50, 100 and 200 µg) of each enzyme preparation. The repeatability results within individual laboratories were satisfactory with coefficients of variation (CVs) ranging from 4 to 8% regardless of the enzyme amount tested. However, the inter-laboratory variability was high (CV > 15%) compared to existing standards for bioanalytical assays. We identified and weighted the contributions to inter-laboratory variability of several parameters associated with the various pH-stat equipment used in this study (e.g. reaction vessel volume and shape, stirring mode and rate, burette volume for the automated delivery of sodium hydroxide). Based on this, we established recommendations for improving the reproducibility of lipase assays using the pH-stat technique. Defining accurate and complete recommendations on how to correctly quantify activity levels of enzyme preparations is a gateway to promising comparison of in vitro data obtained from different laboratories following the same in vitro digestion protocol.
AB - In vitro digestion studies often use animal digestive enzyme extracts as substitutes of human gastric and pancreatic secretions. Pancreatin from porcine origin is thus commonly used to provide relevant pancreatic enzymes such as proteases, amylase and lipase. Rabbit gastric extracts (RGE) have been recently introduced to provide gastric lipase in addition to pepsin. Before preparing simulated gastric and pancreatic extracts with targeted enzyme activities as described in in vitro digestion protocols, it is important to determine the activities of enzyme preparations using validated methods. The purpose of this inter-laboratory study within the INFOGEST network was to test the repeatability and reproducibility of lipase assays using the pH-stat technique for measuring the activities of gastric and pancreatic lipases from various sources. Twenty-one laboratories having different pH-stat devices received the same protocol with identical batches of RGE and two pancreatin sources. Lipase assays were performed using tributyrin as a substrate and three different amounts (50, 100 and 200 µg) of each enzyme preparation. The repeatability results within individual laboratories were satisfactory with coefficients of variation (CVs) ranging from 4 to 8% regardless of the enzyme amount tested. However, the inter-laboratory variability was high (CV > 15%) compared to existing standards for bioanalytical assays. We identified and weighted the contributions to inter-laboratory variability of several parameters associated with the various pH-stat equipment used in this study (e.g. reaction vessel volume and shape, stirring mode and rate, burette volume for the automated delivery of sodium hydroxide). Based on this, we established recommendations for improving the reproducibility of lipase assays using the pH-stat technique. Defining accurate and complete recommendations on how to correctly quantify activity levels of enzyme preparations is a gateway to promising comparison of in vitro data obtained from different laboratories following the same in vitro digestion protocol.
KW - Enzyme activity
KW - INFOGEST
KW - Inhibitor
KW - Lipases
KW - Lipolysis
KW - Titration method
U2 - 10.1016/j.jff.2021.104497
DO - 10.1016/j.jff.2021.104497
M3 - Journal article
AN - SCOPUS:85105584594
VL - 82
JO - Journal of Functional Foods
JF - Journal of Functional Foods
SN - 1756-4646
M1 - 104497
ER -
ID: 273634235