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RECIPROCAL INTERACTIONS BETWEEN RED RASPBERRY POLYPHENOLS AND GUT MICROBIOME COMPOSITION AND METABOLIC HEALTH
Title
RECIPROCAL INTERACTIONS BETWEEN RED RASPBERRY POLYPHENOLS AND GUT MICROBIOME COMPOSITION AND METABOLIC HEALTH
Creator
Zhang, Xuhuiqun
Date
2020
Description
Red raspberries (RRB) and fructo-oligosaccharides (FOS) have been associated with reduced risk of developing cardio-metabolic diseases. RRB...
Show moreRed raspberries (RRB) and fructo-oligosaccharides (FOS) have been associated with reduced risk of developing cardio-metabolic diseases. RRB are uniquely high in anthocyanin- and ellagitannin- type (poly)phenols, however, these (poly)phenols have low bioavailability. Gut microbiota can improve (poly)phenol bioavailability through fermentation processes generating absorbable metabolites and altering gut microbiota structure. However, clinical evidence on the effects of RRB intake on the gut microbiome, (poly)phenolic metabolites and metabolic health is lacking. Further, fermentable carbohydrates (FOS) that selectively stimulate gut microbiota growth may enhance metabolite generation. Therefore, the aim of this research is to investigate the interactions between the gut microbiome and RRB, and explore added effects of FOS as a possible nutritional strategy for improving metabolic health of at risk individuals with prediabetes and insulin resistance (PreDM-IR). Through a series of investigations drawn from a randomized clinical trial (RCT), the following hypotheses were tested: 1) Individuals with PreDM-IR will have a distinctive gut microbiome, and lower capacity to metabolize (poly)phenols compared to healthy individuals; 2) RRB intake for 4-week will increase microbial-derived (poly)phenolic metabolites and adding FOS will augment the effect; 3) RRB intake will improve metabolic risk factors in PreDM-IR and adding FOS will augment the RRB effect; 4) RRB and RRB+FOS supplementations will alter the structure of the gut microbiome explaining variances observed in metabolites and metabolic outcomes. In this single-blinded, crossover RCT, adults with PreDM-IR (n=26) and a healthy group (n=10) consumed 1 cup RRB (fresh weight equivalence) per day or RRB with 8g FOS per day for 4 weeks in random order separated by 4-week washout. Metabolic risk factors, (poly)phenolic metabolites and metagenomic profile were assessed before and after supplementation. Baseline characterization before supplementation revealed distinctive metabolites and metagenomics profiles related to metabolic status. After 4-week RRB, microbial (poly)phenolic metabolites, metabolic health indices and gut microbiome structure beneficially shifted in PreDM-IR group. Adding FOS increased specific microbial species and phenolic metabolites that correlated with β-cell function in PreDM-IR. Overall, nutritional strategies incorporating RRB and FOS may improve metabolic health of individuals with PreDM-IR through modulating gut microbiome composition and the capacity to metabolize RRB (poly)phenols.
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CHARACTERIZATION OF HERBS AND SPICES PHYTOCHEMICALS AND PHARMACOKINETIC PROFILE OVER 24-HOUR AFTER CONSUMPTION IN OVERWEIGHT/OBESE ADULTS
Title
CHARACTERIZATION OF HERBS AND SPICES PHYTOCHEMICALS AND PHARMACOKINETIC PROFILE OVER 24-HOUR AFTER CONSUMPTION IN OVERWEIGHT/OBESE ADULTS
Creator
Huang, Yudai
Date
2022
Description
The health benefits of herbs and spices (H/S) have been known since ancient times. They are a rich source of phytochemicals, such as phenolic...
Show moreThe health benefits of herbs and spices (H/S) have been known since ancient times. They are a rich source of phytochemicals, such as phenolic compounds and terpenoids. However, there is limited information on their absorption and metabolism in humans. Therefore, the primary objective of this study is to identify and characterize phytochemical compounds in H/S mixtures and their absorption and metabolism in the human body over 24 h. H/S and plasma samples used in this study were from a randomized, single-blinded, 4-arm, 24 h, multi-sampling, single-center crossover clinical trial (Clincaltrials.gov NCT03926442) conducted in obese or overweight adults (n=24, aged 37 ± 3 years, BMI=28.4 ± 0.6 kg/m2). Plasma samples were collected at baseline (t=0 h), 0.5, 1, 2, 4, 5.5, 7, and 24 h after consuming a high-fat high-carbohydrate (HFHC) meal with salt and pepper (control) or the control meal with 6 g of three different H/S mixtures (Italian herb: rosemary, basil, thyme, oregano, and parsley in the same ratio; cinnamon; and pumpkin pie spice containing cinnamon, ginger, nutmeg and allspice, the ratio unknown). The phytochemical compounds in the H/S mixtures and their metabolites in human plasma were tentatively identified and quantified by dynamic multiple reaction monitoring transitions on UHPLC-QQQ-MS/MS. Statistical analysis was conducted on SAS-PC 9.4 using non-parametric test via NPAR1WAY procedure. A total of 79 phytochemical compounds were quantified from samples of three H/S mixtures and pepper, of which 36 were flavonoids conpounds, 8 were terpenoids, 27 phenolic acids, and 9 were identified as other compounds. Acetone showed the highest extraction ability for both (poly)phenols and terpenoids in H/S compared to other organic solvents (50% and 80% methanol, ethyl acetate and chloroform). Italian herb contains 763.1 mg/100 g flavonoids, 879 mg/100 g phenolic acids, and 498.6 mg/100g terpenoids; cinnamon contains 981 mg/100 g flavonoids, 11.2 mg/100g phenolic acids, 292.3 mg/100g coumarin, and 1977.1 mg/100 g cinnamaldehyde; pumpkin pie spice contains 655.8 mg/100 g flavonoids, 17.1 mg/100 g phenolic acids, 226.5 mg/100 g coumarin, and 1633 mg/100 g cinnamaldehyde. A total of 47 metabolites were tentatively identified and quantified in plasma samples after H/S consumption over 24 h. Plasma concentrations of carnosic acid derivatives and the glucuronidation products increased after intake of Italian herb, and the Area under the curve (AUC0-24h) was significantly different from control (all P < 0.05) except carnosol glucuronide. Carnosic acid and carnosol had Tmax at 3.4±1.1 and 1.8±0.3 h, respectively, while both of their conjugated glucuronides kept increasing until 24 h. Coumarin glucuronide was increased by cinnamon and pumpkin pie spice consumption with peak concentrations reached at between 1.5-1.6 h. The AUC0-24h after both meals were significantly different from control meal, both P < 0.05. Our results suggest that H/S contain diverse categories of phytochemical compounds that are absorbed and metabolized in the human body into various metabolites in response to 3 different H/S test meals and their appearance in the blood starts as early as around 0.5 h and extends to as long as 24 h for select metabolites.
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AN IMPROVED VALIDATED METHOD FOR THE DETERMINATION OF SHORT-CHAIN FATTY ACIDS IN HUMAN FECAL SAMPLES BY GC-FID
Title
AN IMPROVED VALIDATED METHOD FOR THE DETERMINATION OF SHORT-CHAIN FATTY ACIDS IN HUMAN FECAL SAMPLES BY GC-FID
Creator
Freeman, Morganne M
Date
2022
Description
Short-chain fatty acids (SCFAs) are metabolites produced by the gut microbiota through the fermentation of non-digestible carbohydrates....
Show moreShort-chain fatty acids (SCFAs) are metabolites produced by the gut microbiota through the fermentation of non-digestible carbohydrates. Recent studies suggest that gut microbiota composition, diet and metabolic status play an important role in the production of SCFAs. Current methods for the analysis of SCFAs are complex and inconsistent between research studies. The primary objective of this study was to develop a simplified method for standardized SCFA analysis in human fecal samples by gas chromatography with flame ionization detection (GC-FID). A secondary objective was to apply the method to fecal samples from a previous randomized, crossover clinical trial comparing participants with pre-diabetes mellitus and insulin resistance (IR-group, n=20) to a metabolically healthy reference group (R-group, n=9) after daily consumption of a red raspberry smoothie (RRB, 1 cup fresh-weight equivalent) with or without fructo-oligosaccharide (RRB + FOS, 1 cup RRB + 8g FOS) over a 4-week intervention period. Extraction parameters, including solvent selection and water content of the sample, were investigated before finalizing the method. Freeze-dried fecal samples (0.5 g) were suspended in 5 mL of milli-Q water, vortexed and centrifuged at 3,214 x g for 10 minutes. The supernatant was transferred to a clean tube, acidified with 5.0 M HCl and centrifuged again at 12,857 x g for 5 minutes. The resulting supernatant was transferred to a GC vial for analysis by GC-FID. Linear regression data for standards at concentrations 5-2000 ppm ranged from 0.99994-0.99998. Limit of detection (LOD) ranged from 0.02-0.23 µg/mL. Limit of quantification (LOQ) ranged from 0.08-0.78 µg/mL. The validated method was then applied to fecal samples collected from a previously conducted study. Nine SCFAs were identified and quantified (acetic, propionic, iso-butyric, butyric, iso-valeric, valeric, 4-methyl valeric, hexanoic and heptanoic acids). Statistical analysis (Student’s t-test, ANCOVA) was performed on PC-SAS 9.4 (SAS Institute). Acetic acid was significantly lower in the IR-group compared to the R-group before starting intervention (baseline, Week 0, IR v R-group, p=0.014). Intervention analysis comparing RRB to RRB + FOS at 4 weeks (WK4) showed a significant difference in 4-methyl valeric acid (p = 0.040) in the R-group. Trends of decreased SCFA content after 4-weeks of RRB and RRB + FOS compared to baseline were observed in both groups, though changes were not significantly different between dietary interventions at 4 weeks (p>0.05). Metabolic status and dietary intervention are discussed in relation to their impact on SCFA content in fecal samples and mechanisms of biological use as a metabolite. Limitations of the study include sample size and using only feces and not other biological samples for SCFAs analysis, which may be considered for future research.
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