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Polyphenols in Swedish berries and pulses: Characterization and in vivo effects of fractions from lingonberries and brown beans in an Apoe-/- mouse model
Linnéuniversitetet, Fakulteten för Hälso- och livsvetenskap (FHL), Institutionen för kemi och biomedicin (KOB). Linnéuniversitetet, Kunskapsmiljöer Linné, Hållbar hälsa.ORCID-id: 0000-0001-8097-3483
2021 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Consumption of berries and pulses can protect against atherosclerosis and other metabolic diseases, by modulating plasma lipid profile, gut microbiota, and microbiota-dependent metabolites. These nutritional benefits are often attributed to high content of polyphenols (flavonoids and phenolic acids) and dietary fiber. In this thesis, methods for quantification of flavonoids in berry and pulse matrices were developed. The methods were used for characterization of polyphenols in Swedish berries and pulses. The nutritional benefits of lingonberries, brown beans, and their polyphenol and fiber fractions, were then investigated in an Apoe-/- mouse model. 

The method developed for flavonoid analysis, based on reversed phase HPLC-UV/MS, and extraction procedures were optimized for the berry and pulse matrices, and used to characterize the flavonoids in four Swedish berry species and polyphenols in eight Swedish pulse species (raw and cooked). The fiber and flavonoid fractions from lingonberries and the fiber fraction from brown beans were isolated and evaluated for their in vivo effects on atherosclerosis, compared with whole lingonberries and brown beans. Lingonberries, brown beans, and the isolated fractions were added to a high-fat diet and fed to Apoe-/- mice for 10.5 weeks, to determine their effects on atherosclerotic plaque amount, plasma lipids, gut microbiota, cecal short-chain fatty acids (SCFAs), and plasma methylamines.

The results showed that the optimized analytical methods were capable of quantifying flavonoids in berries and pulses with high precision and recovery. Swedish berries and pulses were found to be good sources of polyphenols. Hydrothermal processing decreased the polyphenol content in pulses by 50-85%, mainly due to leaching into the processing water. In the study on mice, the whole lingonberry diet resulted in 50% lower atherosclerotic plaque amount, but not positively affect the lipid profile. The lingonberry fiber fraction diet doubled the butyric acid concentration in cecum contents and decreased growth of Akkermansia in gut microbiota. Compared with the high-fat control diet, whole bean and its fiber fraction stimulated growth of Bifidobacterium and formation of cecal SCFAs. The diets with whole bean, its fiber fraction, and the two fractions from lingonberry, but not the diet containing whole lingonberry, resulted in higher plasma trimethylamine N-oxide (TMAO) concentrations in the Apoe-/- mice. 

On comparing the diets, it was concluded that the whole lingonberry, but not the lingonberry flavonoid and fiber fractions, resulted in less atherosclerotic plaques. Whole brown and the bean fiber fraction showed a tendency for less atherosclerotic plaques. The effects on plasma lipids, gut microbiota, and microbiota-dependent metabolites in the Apoe-/- mouse model were mainly caused by flavonoids in the case of lingonberries and by fiber in the case of brown beans.

Abstract [sv]

Konsumtionen av bär och baljväxter har visat sig motverka ateroskleros (åderförkalkning) och andra metabola sjukdomar genom att påverka plasmalipidprofilen, tarmens mikrobiota och mikrobiota-beroende metaboliter. De näringsmässiga fördelarna hos bär och baljväxter tillskrivs ofta det höga innehållet av polyfenoler (flavonoider och fenolsyror) och kostfiber. Syftet med denna avhandling var att utveckla metoder för kvantifiering och karaktärisering av polyfenoler i bär och baljväxter, samt att utvärdera effekten på Apoe-/- möss när de utfordras med en kost berikad med lingon och bruna bönor samt polyfenol- respektive fiberfraktioner från dessa livsmedel.

Metoder för analys av vissa flavonoider baserade på omvänd-fas HPLC-UV-MS anpassades, och extraktionsförfarandena optimerades för bär- och baljväxter. Flavonoiderna i svenska bär (lingon, hallon, blåbär och jordgubbar) karaktäriserades och sammansättningen av både flavonoider och fenolsyror studerades i åtta svenska bönor och ärtor (torkade respektive kokta). Vidare isolerades fiber- och flavonoidfraktionerna från lingon och fiberfraktionen från bruna bönor för att utvärderas - i jämförelse med hela lingon och bruna bönor - beträffande deras effekt in vivo på ateroskleros. Tillsatserna kombinerades med en fettrik kost till Apoe-/- möss i 10,5 veckor för att bestämma deras effekt på aterosklerotisk plackmängd, plasmalipider, tarm-mikrobiota, cekala korta fettsyror (SCFAs), plasmakreatinin och plasmametylaminer.

Resultaten visade att de optimerade analysmetoderna var lämpliga för kvantifiering av polyfenoler i bär och baljväxter med hög precision och högt utbyte. Svenska bär och baljväxter visade sig vara bra källor till polyfenoler. Kokning minskade polyfenolhalten i baljväxter med 50–85 %, främst på grund av urlakning i processvattnet. Data från djurstudien visade att en diet berikad med lingon resulterade i 50% lägre aterosklerotisk plackmängd, men att lipidprofilen inte påverkades. Lingonfiberfraktionen fördubblade halten smörsyra i caecum och minskade tillväxten av Akkermansia i tarm-mikrobiotan. Hela bruna bönor och dess fiberfraktion stimulerade både tillväxten av Bifidobacterium och bildandet av cecal SCFA (korta fettsyror). Alla berikningar, förutom hela lingon, resulterade i förhöjda nivåer av trimetylamin N-oxid (TMAO) i plasma hos Apoe-/-mössen, jämfört med en kontrolldiet utan berikning.

Vid jämförelse av dieterna visade det sig att hela lingon, men inte lingonflavonoidfraktion och fiberfraktion, resulterade i mindre aterosklerotiska plack. Hela bruna bönor och bönfiberfraktionen visade en tendens till mindre aterosklerotiska plack. Effekterna på plasmalipider, tarm-mikrobiota och mikrobiotaberoende metaboliter i Apoe-/- mössen orsakades främst av flavonoider när det gäller lingon och av fibrer när det gäller bruna bönor.

sted, utgiver, år, opplag, sider
Växjö: Linnaeus University Press, 2021. , s. 74
Serie
Linnaeus University Dissertations ; 432
Emneord [en]
lingonberry, brown bean, flavonoids, phenolic acids, dietary fiber, HPLC-UV/MS, atherosclerotic plaque, plasma lipid, gut microbiota, short-chain fatty acids, trimethylamine N-oxide, Apoe-/- mice
Emneord [sv]
lingon, bruna bönor, flavonoider, fenolsyror, kostfiber, HPLCUV/ MS, aterosklerotisk plaketter, plasmalipid, tarm-mikrobiota, kortkedjiga fettsyror, trimetylamin N-oxid, Apoe-/- möss
HSV kategori
Forskningsprogram
Naturvetenskap, Livsmedelsvetenskap
Identifikatorer
URN: urn:nbn:se:lnu:diva-108786ISBN: 9789189460447 (tryckt)ISBN: 9789189460454 (digital)OAI: oai:DiVA.org:lnu-108786DiVA, id: diva2:1624343
Disputas
2021-12-17, Lapis, building Vita, Kalmar, 09:00 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2022-01-04 Laget: 2022-01-04 Sist oppdatert: 2024-03-05bibliografisk kontrollert
Delarbeid
1. Characterization of Flavonoid Compounds in Common Swedish Berry Species
Åpne denne publikasjonen i ny fane eller vindu >>Characterization of Flavonoid Compounds in Common Swedish Berry Species
2020 (engelsk)Inngår i: Foods, E-ISSN 2304-8158, Vol. 9, nr 3, s. 1-13, artikkel-id 358Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Berries are considered an ideal source of polyphenols, especially from the flavonoid group. In this study, we examined the flavonoid content in 16 varieties of Swedish lingonberry, raspberry, blueberry, and strawberry. Nineteen flavonoids were simultaneously quantified using external standards. An additional 29 flavonoids were tentatively identified using MS as no standards were available. Quantification was done using HPLC-UV after optimization of chromatographic and extraction procedures. The method showed high linearity within the range of 2–100 µg/mL (correlation co-efficient >0.999), intra- and inter-day precision of 1.7–7.3% and average recovery above 84% for all compounds. Blueberries and lingonberries were found to contain higher contents of flavonoids (1100 mg/100 g dry weight) than raspberries and strawberries (500 mg/100 g dry weight). Anthocyanins were the dominant flavonoids in all berries. The tentatively characterized compounds contribute 18%, 29%, 61%, and 67% of the total flavonoid content in strawberries, lingonberries, raspberries, and blueberries, respectively. Overall, Swedish berries were shown to be good sources of polyphenols.

sted, utgiver, år, opplag, sider
MDPI, 2020
HSV kategori
Forskningsprogram
Naturvetenskap, Livsmedelsvetenskap
Identifikatorer
urn:nbn:se:lnu:diva-93494 (URN)10.3390/foods9030358 (DOI)000526529800045 ()32204535 (PubMedID)2-s2.0-85082777471 (Scopus ID)
Tilgjengelig fra: 2020-04-16 Laget: 2020-04-16 Sist oppdatert: 2023-04-20bibliografisk kontrollert
2. Phenolic compounds in Swedish dried pulses: Characterization, retention and distribution during hydrothermal treatment processes
Åpne denne publikasjonen i ny fane eller vindu >>Phenolic compounds in Swedish dried pulses: Characterization, retention and distribution during hydrothermal treatment processes
2021 (engelsk)Inngår i: Journal of Food Composition and Analysis, ISSN 0889-1575, E-ISSN 1096-0481, Vol. 102, artikkel-id 104056Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Pulses are an important crop for the human diet. The aim of this work was to characterize phenolic compounds (flavonoids and phenolic acids) in common Swedish dried pulses and to investigate the retention and distribution of phenolic compounds during hydrothermal treatment processes, including soaking, draining, and boiling. An ultrasound-assisted extraction method was optimized for flavonoid extraction. The phenolic compounds in the raw and cooked pulses, and the processing (soaking and boiling) water were analyzed using HPLC-UV/MS. Optimized extraction conditions for flavonoids were: 70 % methanol, pH 2, and sonication for 60 min. A total of 33 flavonoids and 6 phenolic acids were directly or tentatively identified and quantified. The profile of phenolic compounds largely depends on the pulse type with the total content ranging between 38-1860 mu g/g dry matter in the raw material. In most pulses, hydrothermal treatment led to a decrease of 50-85 % of total phenolic content, mainly due to leaching. The distribution of major phenolic compound subgroups in the soaking water, boiling water, and cooked pulse varied. The rate of retention of individual phenolic compounds depends possibly on both their structure and the pulse matrix.

sted, utgiver, år, opplag, sider
Elsevier, 2021
Emneord
Swedish pulses, Hydrothermal treatment, Phenolic acids, Flavonoids, Extraction, HPLC-UV, MS
HSV kategori
Forskningsprogram
Naturvetenskap, Livsmedelsvetenskap
Identifikatorer
urn:nbn:se:lnu:diva-106777 (URN)10.1016/j.jfca.2021.104056 (DOI)000687397700004 ()2-s2.0-85111008118 (Scopus ID)2021 (Lokal ID)2021 (Arkivnummer)2021 (OAI)
Tilgjengelig fra: 2021-09-03 Laget: 2021-09-03 Sist oppdatert: 2023-04-20bibliografisk kontrollert

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