Development of a reliable pH-STAT <i>in-vitro</i> model for gastrointestinal digestion of lipids and application for infant formula

WANG, Mengxun; XU, Shixiang; CHEONG, Ling-Zhi; XU, Xuebing; BI, Yanlan; ZHANG, Hong

doi:10.1590/fst.115221

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Food Science and Technology

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Original Article • Food Sci. Technol 42 • 2022 • https://doi.org/10.1590/fst.115221 copy

Development of a reliable pH-STAT in-vitro model for gastrointestinal digestion of lipids and application for infant formula

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Present study aims to establish a reliable infant digestion model using pH-STAT method and the model is also applied to static in-vitro digestion of different infant formulas. Model-3 which consisted of NaCl, KCl and CaCl₂ as gastrointestinal digestive fluids demonstrated better repeatability than model-1 was selected for the evaluation of static in-vitro digestion of different infant formulas (different fatty acids, protein composition and with/without milk fat globule membrane) using pH-STAT method. Three infant formulas (IF) with different fatty acids composition were evaluated and IF2 which contained the highest amounts of medium-chain saturated fatty acids (8.02 ± 0.47%) demonstrated the highest in-vitro gastrointestinal lipolysis rate (65.88 ± 0.24%). Infant formulas containing peptides and amino acids were more easily hydrolyzed by protease, which probably enhanced the release of FFA (IF1, 50.60 ± 0.87%; IF4, 60.17 ± 2.97%; IF5, 62.21 ± 2.27%; IF6, 66.51 ± 2.38%). In addition, the presence of milk fat globule membrane (MFGM, IF7) was found to accelerate in-vitro gastrointestinal lipolysis (IF1, 50.60 ± 0.87%; IF7, 58.05 ± 1.77%). In summary, the new developed model is suitable for investigating in-vitro digestion of different infant formulas.

Keywords:
pH-STAT; simulated digestive fluids; in-vitro gastrointestinal lipolysis; fatty acid composition; protein composition; MFGM

Infant Formulas	FAC¹ 1 FAC: Fatty acid composition; MFGM: Milk fat globular membrane.	Protein composition	MFGM protein
IF1	Cow	Standard	None
IF2	Goat	Standard	None
IF3	Soybean	Standard	None
IF4	Cow	Partially	None
IF5	Cow	Extensively	None
IF6	Cow	Amino acid	None
IF7	Cow	Standard	MFGM

	IF1	IF2	IF3	IF4	IF5	IF6	IF7
Energy (KJ)	269	268	277	284	250	283	284
Fat (g)	3.25	3.13	3.5	3.7	3.18	3.51	3.65
Carbohydrate (g)	7.25	7.43	6.70	7.10	6.18	7.16	7.29
Protein (g)	1.47	1.50	1.7	1.54	1.68	1.88	1.43

Model-1 (Cheong et al., 2018Cheong, L.-Z., Jiang, C., He, X., Song, S., & Lai, O.-M. (2018). Lipid profiling, particle size determination, and in vitro simulated gastrointestinal lipolysis of mature human milk and infant formula. Journal of Agricultural and Food Chemistry, 66(45), 12042-12050. http://dx.doi.org/10.1021/acs.jafc.8b03998. PMid:30362342. http://dx.doi.org/10.1021/acs.jafc.8b039... )	Simulated gastric fluid	NaTC	NaCl	Tris	Maleic acid	phospholipid
	Simulated gastric fluid	80 μM	68 mM	2 mM	2 mM	2 μM
	Simulated intestinal fluid	NaTC	NaCl	Tris	Maleic acid	phospholipid
	Simulated intestinal fluid	2 mM	150 mM	2 mM	2 mM	0.18 mM
Model-2 (Ménard et al., 2018Ménard, O., Bourlieu, C., De Oliveira, S. C., Dellarosa, N., Laghi, L., Carrière, F., Capozzi, F., Dupont, D., & Deglaire, A. (2018). A first step towards a consensus static in vitro model for simulating full-term infant digestion. Food Chemistry, 240, 338-345. http://dx.doi.org/10.1016/j.foodchem.2017.07.145. PMid:28946281. http://dx.doi.org/10.1016/j.foodchem.201... )	Simulated gastric fluid	NaCl	KCl
	Simulated gastric fluid	94 mM	13 mM
	Simulated intestinal fluid	NaCl	KCl	NaHCO₃	CaCl₂
	Simulated intestinal fluid	164 mM	10 mM	85 mM	3.1 mM
Model-3 (Mat et al., 2016Mat, D., Le Feunteun, S., Michon, C., & Souchon, I. (2016). In vitro digestion of foods using pH-stat and the INFOGEST protocol: Impact of matrix structure on digestion kinetics of macronutrients, proteins and lipids. Food Research International, 88, 226-233. http://dx.doi.org/10.1016/j.foodres.2015.12.002. http://dx.doi.org/10.1016/j.foodres.2015... ; Ménard et al., 2018Ménard, O., Bourlieu, C., De Oliveira, S. C., Dellarosa, N., Laghi, L., Carrière, F., Capozzi, F., Dupont, D., & Deglaire, A. (2018). A first step towards a consensus static in vitro model for simulating full-term infant digestion. Food Chemistry, 240, 338-345. http://dx.doi.org/10.1016/j.foodchem.2017.07.145. PMid:28946281. http://dx.doi.org/10.1016/j.foodchem.201... )	Simulated gastric fluid	NaCl	KCl
	Simulated gastric fluid	94 mM	13 mM
	Simulated intestinal fluid	NaCl	KCl	NaCl	CaCl₂
	Simulated intestinal fluid	164 mM	10 mM	85 mM	3.1 mM

Model	The Volume of Digestive fluids	Enzyme concentrations
Model-1	12% of meal, 28% of gastric secretion and 60% of intestinal secretion	Gastric lipase: 60U/mL
		Pepsin: 450 U/mL
		Pancreatic: 300 USP/mL
Model-2	39% of meal, 23% of gastric secretion and 38% of intestinal secretion	Gastric lipase: 19U/mL
Model-2		Pepsin: 268 U/mL
Model-3		Pancreatic lipase: 90/mL

	IF1	IF2	IF3
C8:0	0.65 ± 0.18^a	5.48 ± 0.37^b	1.44 ± 0.16^a
C10:0	1.20 ± 0.06^a	2.55 ± 0.09^b	1.21 ± 0.01^a
C12:0	5.06 ± 0.19^a	4.74 ± 0.13^a	10.5 ± 0.41^b
C14:0	4.93 ± 0.01^a	4.39 ± 0.12^b	4.31 ± 0.01^b
C14:1	0.40 ± 0.02^a	0.16 ± 0.01^b	ND
C15:0	0.35 ± 0.01^a	0.25 ± 0.01^b	ND
C16:0	19.75 ± 0.05^a	18.40 ± 0.53^b	17.21 ± 0.09^c
C16:1	0.46 ± 0.01^a	0.31 ± 0.01^b	0.12 ± 0.01^c
C17:0	0.21 ± 0.01^a	0.21 ± 0.01^a	ND
C17:1	0.12 ± 0.01^a	0.12 ± 0.01^a	ND
C18:0	4.79 ± 0.05^a	5.06 ± 0.16^a	3.66 ± 0.04^b
C18:1	32.63 ± 0.07^a	32.33 ± 0.96^a	40.61 ± 0.28^b
C18:1t	3.66 ± 0.18^a	2.98 ± 0.05^b	ND
C18:2	22.10 ± 0.13^a	21.07 ± 0.65^a	18.38 ± 0.12^b
C18:2t	ND	0.13 ± 0.01	ND
C18:3t	3.02 ± 0.04^a	2.80 ± 0.09^b	1.61 ± 0.06^c
C20:1	0.58 ± 0.02^a	0.60 ± 0.04^a	0.15 ± 0.01^b
C20:3n3	ND	0.59 ± 0.02^a	0.53 ± 0.1^a
C22:6n3	0.19 ± 0.01^a	0.17 ± 0.01^b	0.24 ± 0.01^c
MC-SFA(C6-10)	1.84 ± 0.24^a	8.02 ± 0.47^b	2.65 ± 0.16^a
SFA	36.92 ± 0.35^a	41.08 ± 1.45^b	38.32 ± 0.44^a
MUFA	37.83 ± 0.25^a	36.5 ± 1.05^a	40.87 ± 0.28^b
PUFA	25.3 ± 0.17^a	24.68 ± 0.69^a	20.76 ± 0.08^b
ratio of saturated to unsaturated fatty acids	0.59 ± 0.007	0.67 ± 0.05	0.62 ± 0.01

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[1] *Corresponding author: zhanghongsh@cn.wilmar-intl.com