Quantification Of Fatty Acids Research Articles

Unraveling the Double Bond Position of Fatty Acids by GC-MS Using Electron Capture APCI and In-Source Fragmentation Patterns.

The position of double bonds in unsaturated fatty acids is strongly connected to their biological effects, but their analytical characterization is still challenging. However, the ionization of unsaturated fatty acids by a GC-APCI leads to regiospecific in-source fragment ions, which can be used to identify the double bond position. The fragment ions are oxidized species that occur mostly at the double bond closest to the carboxylic acid group. This effect can be further promoted by using benzaldehyde as a gas-phase reactant. This allows the identification of the Δ-notation of the fatty acid, and based on additional information such as m/z and retention time, it is possible to annotate the corresponding fatty acid. The developed method also enables the quantification of fatty acids in one step with high selectivity and sensitivity. Moreover, rare fatty acids can be identified in suspected target approaches that are often not available as standards. This was demonstrated by analyzing fish oil samples that provide a complex mixture of highly unsaturated fatty acids and by identifying rare fatty acids such as hexadecatetraenoic acid (FA 16:4 Δ6).

Strategy for characterization and quantification of fatty acids in plasma by parallel d0/d6-dansylhydrazine derivatization combined with UPLC–MS/MS

Fatty acids (FAs) play a vital physiological role in lipid metabolism, which is reported as potential diagnostic biomarker for various diseases. Thus, it is urgent to develop a credible method that can profile FA metabolism with a holistic view. Here, a targeted strategy to screen FAs was developed by parallel labeling with d0/d6-dansylhydrazine (d0/d6-DnsHz) and using ultra-high performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC–MS/MS) in data-dependent MS/MS (ddMS2) mode. The simple and mild derivatization procedure within 3 h allowed for a significant improvement in sensitivity. Additionally, the characteristic product ions introduced by the derivatization reagent assist to identify the unknown FA species. A quantitation method was established by multiple reaction monitoring (MRM) and the d6-DnsHz tagged standards for each analyte were used as internal standards to overcome the matrix effects. By applying the method to determine FA levels in plasma collected from the esophageal squamous cell carcinoma (ESCC) patients and healthy controls, 65 FA metabolites were characterized and six FAs were found to be altered by the invasion of tumors. The parallel derivatization strategy provides insights into the identification of unknown FAs and paves a new way for targeted metabolomics. Also, this novel method is a powerful tool for characterization and quantification of FAs in biological samples, which shows a great potential application in clinical diagnosis and investigation of disease mechanisms.

Insect-based aquafeeds modulate the fatty acid profile of zebrafish: A comparison on the different life stages

Insects are considered as an alternative and sustainable ingredient for feed production. In this study, Hermetia illucens (Hi) prepupae (fed on coffee roasted by-product added with 10 % Schyzochytrium sp., a marine protist rich in polyunsaturated fatty acids) was processed into meal and included at graded levels in five experimental diets to replace dietary fish meal (0, 25, 50, 75, 100 %) offered to zebrafish (Danio rerio) as experimental model. The fatty acids (FAs) profile of fish larvae (20 days), juveniles (2 months, deprived of the viscera) and adults (6 months, male and female, deprived of the viscera) specimens was investigated to evaluate the impact of dietary FM replacement with full-fat Hi prepupae meal. For the first time, the quantification of FAs in Danio rerio, performed by gas chromatography-mass spectrometry, was computed in absolute terms (mg 100 g−1 dw), identifying the real variation in the content of any single FA. In fish, quantified total FAs in larvae, juvenile, adult male and adult female were 134 ± 2, 235 ± 8, 266 ± 3 and 266 ± 8 mg g−1 dw, respectively. With respect to zebrafish fed the control diet, specimens fed diet with increasing level of Hi prepupae meal showed significantly higher content of saturated and omega6 FAs, and significantly lower content of poly-unsaturated and omega3 FAs, reflecting partially the FAs composition of the administered diets. At the same time, the docosahexaenoic/eicosapentaenoic acid ratio increased significantly. Moreover, adult female showed a higher content of PUFA with respect to adult male, which could be justified by the needs of physiological reproductive processes. The principal component analysis demonstrated that the FAs composition in Danio rerio depends on both the life stage and the diet.

Development and validation of derivatization-based LC-MS/MS method for quantification of short-chain fatty acids in human, rat, and mouse plasma

Short-chain fatty acids (SCFAs), the end products of gut microbial fermentation of dietary fibers and non-digestible polysaccharides, act as a link between the microbiome, immune system, and inflammatory processes. The importance of accurately quantifying SCFAs in plasma has recently emerged to understand their biological role. In this work, a sensitive and reproducible LC-MS/MS method is reported for SCFAs quantification in three different matrices such as human, rat and mouse plasma via derivatization, using as derivatizing agent O-benzylhydroxylamine (O-BHA), coupled with liquid-liquid extraction. First, the instrumental parameters of the mass spectrometer and then the chromatographic conditions were optimized using previously SCFAs derivatives synthetized and used as standards. After that, the best conditions for derivatization and extraction from plasma were studied and a series of determinations were performed on human, rat, and mouse plasma aliquots to validate the overall method (derivatization, extraction, and LC-MS/MS determination). The method showed good performance in terms of recovery (> 80%), precision (RSD <14%), accuracy (RE < ± 10%) and sensitivity (LOQ of 0.01 µM for acetic, butyric, propionic and isobutyric acid) in all plasma samples. The method thus developed and validated was applied to the quantification of major SCFAs in adult and aged mice, germ-free mice and in germ-free recipient mice subjected to fecal transplant from adult and aged donors. Results highlighted how plasma concentrations of SCFAs are correlated with age further highlighting the importance of developing a method that is reliable for the quantification of SCFAs to study their biological role.

Biochemical Changes Induced by the Administration of Cannabis sativa Seeds in Diabetic Wistar Rats.

The present pilot study investigates the blood biochemical changes induced by hemp seeds in rats with diabetes. The composition of industrial hemp seeds, antioxidant activity, identification and quantification of phenols and fatty acids from hemp oil were determined. The Wistar adult rats used in the experiment were divided into three groups (n = 6) and kept under standard conditions. Group one, the control group (individuals without diabetes), and group two (diabetic individuals) received water and normal food ad libitum, while the third group, also including diabetic individuals, received specific food (hemp seeds) and water ad libitum. Subsequent blood biochemical parameters were determined. Hemp seeds had higher phenol (14 compounds), flavonoids and PUFA contents compared to other plants seeds. In addition, the antioxidant activity in Cannabis sativa was also increased. Moreover, the ratio between n-6 and n-3 was 4.41, ideal for different diseases. Additionally, all biochemical parameters showed significant changes following the treatment. It was shown that high doses of hemp seeds decreased diabetes-induced biochemical damage in rats most probably due to the high content of active compounds. In order to use these seeds in humans, it is essential to find out which hemp compounds are particularly responsible for these effects. Moreover, for the objective investigation of their effects, longer-term studies are needed.

Low-diversity microbiota and an increased metabolism of arginine and aromatic amino acids: a hallmark of hepatic encephalopathy in western Mexican patients with alcohol-associated cirrhosis.

To evaluate the composition and functions of the gut microbiota in patients with decompensated alcohol-associated cirrhosis, with and without hepatic encephalopathy (HE). Faecal samples from 31 inpatients (20 with HE, 11 without HE), and from 18 age-balanced healthy controls (HC), were included. Microbial composition was determined by 16S rRNA amplicon sequencing and analysed using QIIME2. Metabolic pathways were inferred by PICRUSt2, and short-chain fatty acids (SCFAs) quantification was performed by gas chromatography. The gut microbiota of patients with HE was characterized by a diminished α-diversity, compared to no-HE (P<0.01) and HC (P<0.001) groups; β-diversity also differed between HE vs no-HE patients (P<0.05), and between HE vs HC (P<0.001). In patients with HE, Escherichia/Shigella, Burkholderiales and Lactobacillales taxa predominated. In contrast, patients without HE were characterized by Veillonella and Bacteroides. Reduced levels of faecal SCFAs in both groups correlated with a depletion of beneficial taxa, such as Ruminococcus or Faecalibacterium. PICRUSt2 analysis showed both an enhanced catabolism of arginine through ammonia-producing pathways and chorismate biosynthesis in HE patients, a key precursor of aromatic amino acids. The gut microbiota of HE patients exhibits a proinflammatory dysbiotic profile, plus metabolic pathways that produce potentially neurotoxic byproducts.

GC/MS quantification of individual fatty acids of selected green leafy vegetable foliage and their biodiesel attributes

The current demand for edible vegetable oil is increasing worldwide, and the development of new sources of high-quality vegetable edible oil is an essential task. There is also a huge demand for biodiesel in domestic and industrial applications, and foliage oils could be a good source for diesel applications. The current study aimed at the identification and quantification of fatty acids from commonly consumed green leafy vegetables (GLVs) viz., Hibiscus cannabinus, Hibiscus sabdariffa, Basella alba, Basella rubra, and Rumex vesicarius and to calculate the biodiesel attributes of the oil. The total oil content was ascertained as the highest in R. vesicarius foliage (3.91 ± 0.27 g/100 g dry leaf powder). GC/MS chromatographic investigation identified 9,12,15-octadecatrienoic acid as a significant compound followed by hexadecanoic acid. In Hibiscus spp. C18:3 (49.3 µmol % and 50.4 µmol %) was recorded to be the most noteworthy followed by C16:0 (23.2 µmol % and 21 µmol %) in H. cannabinus and H. sabdariffa, respectively. The GLVs foliage-fatty acid biodiesel attributes were additionally assessed through an empirical formula. Consequently, the overall examined results will be helpful for the investigation of these oils as vegetable oil for human consumption and biodiesel applications.

Perturbations in fatty acid metabolism and collagen production infer pathogenicity of a novel MBTPS2 variant in Osteogenesis imperfecta.

Osteogenesis imperfecta (OI) is a heritable and chronically debilitating skeletal dysplasia. Patients with OI typically present with reduced bone mass, tendency for recurrent fractures, short stature and bowing deformities of the long bones. Mutations causative of OI have been identified in over 20 genes involved in collagen folding, posttranslational modification and processing, and in bone mineralization and osteoblast development. In 2016, we described the first X-linked recessive form of OI caused by MBTPS2 missense variants in patients with moderate to severe phenotypes. MBTPS2 encodes site-2 protease, a Golgi transmembrane protein that activates membrane-tethered transcription factors. These transcription factors regulate genes involved in lipid metabolism, bone and cartilage development, and ER stress response. The interpretation of genetic variants in MBTPS2 is complicated by the gene's pleiotropic properties; MBTPS2 variants can also cause the dermatological conditions Ichthyosis Follicularis, Atrichia and Photophobia (IFAP), Keratosis Follicularis Spinulosa Decalvans (KFSD) and Olmsted syndrome (OS) without skeletal abnormalities typical of OI. Using control and patient-derived fibroblasts, we previously identified gene expression signatures that distinguish MBTPS2-OI from MBTPS2-IFAP/KFSD and observed stronger suppression of genes involved in fatty acid metabolism in MBTPS2-OI than in MBTPS2-IFAP/KFSD; this was coupled with alterations in the relative abundance of fatty acids in MBTPS2-OI. Furthermore, we observed a reduction in collagen deposition in the extracellular matrix by MBTPS2-OI fibroblasts. Here, we extrapolate our observations in the molecular signature unique to MBTPS2-OI to infer the pathogenicity of a novel MBTPS2 c.516A>C (p.Glu172Asp) variant of unknown significance in a male proband. The pregnancy was terminated at gestational week 21 after ultrasound scans showed bowing of femurs and tibiae and shortening of long bones particularly of the lower extremity; these were further confirmed by autopsy. By performing transcriptional analyses, gas chromatography-tandem mass spectrometry-based quantification of fatty acids and immunocytochemistry on fibroblasts derived from the umbilical cord of the proband, we observed perturbations in fatty acid metabolism and collagen production similar to what we previously described in MBTPS2-OI. These findings support pathogenicity of the MBTPS2 variant p.Glu172Asp as OI-causative and highlights the value of extrapolating molecular signatures identified in multiomics studies to characterize novel genetic variants.

A chemical study of yoghurt produced under isostatic pressure during storage

Yoghurt fermented under sub-lethal high pressure (10, 20, 30 and 40 MPa at 43 °C), and afterward placed under refrigeration (4 °C for 23 days) was studied and compared with yoghurt fermented at atmospheric pressure (0.1 MPa). For a deeper analysis, metabolite fingerprinting by nuclear magnetic resonance (NMR), sugars and organic acids assessment by high performance liquid chromatography (HPLC), total fatty acids (TFA) determination and quantification by gas chromatography with a flame ionization detector (GC-FID) were performed. Metabolomic analyses revealed that only 2,3-butanediol, acetoin, diacetyl and formate vary with the increase of pressure and probable relation with pressure influenced diacetyl reductase, acetoin reductase and acetolactate decarboxylase. Yoghurts fermented at 40 MPa had the lowest content in lactose (39.7 % of total sugar reduction) and the less content in TFA (56.1 %). Further research is of interest to understand more about fermentation processes under sub-lethal high pressure.

HPLC-DAD Development and Validation Method for Short-Chain Fatty Acids Quantification from Chicken Feces by Solid-Phase Extraction

A solid-phase extraction (SPE) process, followed by an HPLC-DAD method, was developed and validated to quantify short-chain fatty acids (SCFAs) and applied to analyze chicken feces samples. This study aimed to report the use of the STRATATM-X-A 96 Well Plate SPE cartridge method as the first step in quantifying SCFAs. A stationary reverse-phase Luna Omega C18 column kept at 40 °C was used, with a gradient elution mobile phase (H2SO4 and Acetonitrile, 98:2), a flow rate of 1.2 mL/min, and detection at 210 nm. A mixture of acetic, propionic, and butyric acid was used as thestandard. The method showed a linear relationship, with a coefficient of determination of R2 = 0.9987, R2 = 0.9985, and R2 = 0.9966 for acetic, propionic, and butyric acid, respectively. Concerning sensitivity, an LOD and LOQ of 0.14, 0.14, 0.14 mg/mL and 0.44, 0.45, 0.43 mg/mL were obtained for acetic, propionic, and butyric acid, respectively. According to the sample analysis, the accuracy was 76.05, 95.60, and 81.56% for acetic, propionic, and butyric acid, respectively. The developed method is simple, fast, linear, sensitive, accurate, precise, and robust for the quantification of SCFAs. This could serve as an alternative to conventional methodologies for the determination of these critical components in the intestinal health of chicken feces.

Simultaneous quantification of fatty acids in serum with dual derivatization by liquid chromatography-tandem mass spectrometry.

Fatty acids have multitudinous biological functions and play a crucial role in many biological processes, but due to poor ionization efficiency and lack of appropriate internal standards, the comprehensive quantification of fatty acids by liquid chromatography-tandem mass spectrometry is still challenging. In this study, a new, accurate, and reliable method for quantifying 30 fatty acids in serum using dual derivatization was proposed. Indole-3-acetic acid hydrazide derivants of fatty acids were used as the internal standard and indole-3-carboxylic acid hydrazide derivants of them were used to quantify. The derivatization conditions were systematically optimized and the method validation results showed good linearity with R2 >0.9942, low detection limit (0.03-0.6nM), precision (1.6%-9.8% for intra-day and 4.6%-14.1% for inter-day), recovery (88.2%-107.2% with relative standard deviation<10.5%), matrix effect (88.3%-105.2% with the relative standard deviation <9.9%) and stability (3.4%-13.8% for fatty acids derivants in 24h at 4°C and 4.2%-13.8% for three freeze-thaw cycles). Finally, this method was successfully applied to quantify fatty acids in serum samples of Alzheimer's patients. In contrast to the healthy control group, nine fatty acids showed a significant increase in the Alzheimer's disease group.

Fusion-based strategy of CSA and mobile NIR for the quantification of free fatty acid in wheat varieties coupled with chemometrics

The use of sensor fusion, a novel method of combining artificial senses, has become increasingly popular in the assessment of food quality. This study employed a combination of the colorimetric sensor array (CSA) and mobile near-infrared (NIR) spectroscopy to predict free fatty acids in wheat flour. In conjunction with a partial least squares model, Low- and mid-level fusion strategies were used for quantification. Accordingly, performance of the built model was evaluated based on higher correlation coefficients between calibration and prediction (RC and RP), lower root mean square error of prediction (RMSEP), and a higher residual predictive deviation (RPD). The mid-level fusion coupled PLS model produced superior data fusion findings, with RC = 0.8793, RMSECV = 7.91 mg/100 g, RP = 0.8747, RMSEP = 6.99 mg/100 g, and RPD = 2.27. The findings of the study suggest that the NIR-CSA fusion approach could be effectively applied to the prediction of free fatty acids in wheat flour.

A fast and sensitive high-throughput assay to assess polysorbate-degrading hydrolytic activity in biopharmaceuticals

Hydrolysis of polysorbate in biopharmaceutical products has been ascribed to the enzymatic activity from trace levels of residual host cell proteins. In recent years, significant efforts to identify the causative enzymes typically used elaborate, material-intensive and time-consuming approaches. Therefore, the lack of fast and sensitive assays to monitor their activity remains a major bottleneck for supporting process optimization and troubleshooting activities where time and sample throughput are crucial constraints. To address this bottleneck, we developed a novel Electrochemiluminescence-based Polysorbase Activity (EPA) assay to measure hydrolytic activities in biotherapeutics throughout the drug substance manufacturing process. By combining the favorable features of an in-house designed surrogate substrate with a well-established detection platform, the method yields fast (∼36 h turnaround time) and highly sensitive readouts compatible with high-throughput testing. The assay capability for detecting substrate conversion in a precise and reliable manner was demonstrated by extensive qualification studies and by employing a number of recombinant hydrolases associated with polysorbate hydrolysis. In addition, high assay sensitivity and wide applicability were confirmed for in-process pool samples of three different antibody products by performing a head-to-head comparison between this method and an established liquid chromatography - mass spectrometry based assay for the quantification of free fatty acids. Overall, our results suggest that this new approach is well-suited to resolve differences in hydrolytic activity through all stages of purification.

From In Vitro to In Vivo: A Rational Flowchart for the Selection and Characterization of Candidate Probiotic Strains in Intestinal Disorders.

Experimental and clinical evidence has demonstrated the potential of probiotic strains in the prevention or treatment of inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS). However, there is little data on what the methodology leading to the identification of such strains should be. In this work, we propose a new flowchart to identify strains with probiotic potential for the management of IBS and IBD, which we tested on a collection of 39 lactic acid bacteria and Bifidobacteria strains. This flowchart included in vitro tests of immunomodulatory properties on intestinal and peripheral blood mononuclear cells (PBMCs), assessment of the barrier-strengthening effect by measuring transepithelial electric resistance (TEER) and quantification of short-chain fatty acids (SCFAs) and aryl hydrocarbon receptor (AhR) agonists produced by the strains. The in vitro results were then combined in a principal component analysis (PCA) to identify strains associated with an anti-inflammatory profile. To validate our flowchart, we tested the two most promising strains identified in the PCA in mouse models of post-infectious IBS or chemically induced colitis to mimic IBD. Our results show that this screening strategy allows the identification of strains with potential beneficial effects on colonic inflammation and colonic hypersensitivity.

Quantification of Free Short-Chain Fatty Acids in Raw Cow Milk by Gas Chromatography-Mass Spectrometry

Free short-chain fatty acids (FSCFAs) are a momentous contributor to the flavor of the raw cow milk. Hence, the purpose of this research was to build an approach for the quantification of 10 FSCFAs in raw cow milk. Raw cow milk samples are acidified by hydrochloric acid ethanol (0.5%) solution pretreatment and then processed on the gas chromatography-mass spectrometry. With the exception of iso C5:0 and anteiso C5:0 co-flux, the remaining eight FSCFAs were effectively separated by chromatography. The methodological validation data revealed that the linear relationship satisfied the assay requirements (coefficient of determination >0.999), the limits of quantification were 0.167 to 1.250 μg mL-1, the recoveries ranged from 85.62% to 126.42%, the coefficients of variation were 1.40~12.15%, and no SCFAs in the triglyceride form were potential degradation, and the precision ranging from 0.56% to 9.09%. Our easy, fast, and robust method successfully determined three FSCFAs in raw cow milk without derivatization. Some characteristic features of FSCFAs have been discovered in raw cow milk such as its higher percentages of C4:0 and C6:0. Our research has provided a very valuable method for the future quality and safety control of raw milk and nutritional studies.

HPLC-MS-MS quantification of short-chain fatty acids actively secreted by probiotic strains.

Short-chain fatty acids (SCFAs) are the main by-products of microbial fermentations occurring in the human intestine and are directly involved in the host's physiological balance. As impaired gut concentrations of acetic, propionic, and butyric acids are often associated with systemic disorders, the administration of SCFA-producing microorganisms has been suggested as attractive approach to solve symptoms related to SCFA deficiency. In this research, nine probiotic strains (Bacillus clausii NR, OC, SIN, and T, Bacillus coagulans ATCC 7050, Bifidobacterium breve DSM 16604, Limosilactobacillus reuteri DSM 17938, Lacticaseibacillus rhamnosus ATCC 53103, and Saccharomyces boulardii CNCM I-745) commonly included in commercial formulations were tested for their ability to secrete SCFAs by using an improved protocol in high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS-MS). The developed method was highly sensitive and specific, showing excellent limits of detection and quantification of secreted SCFAs. All tested microorganisms were shown to secrete acetic acid, with only B. clausii and S. boulardii additionally able to produce propionic and butyric acids. Quantitative differences in the secretion of SCFAs were also evidenced. The experimental approach described in this study may contribute to the characterization of probiotics as SCFA-producing organisms, a crucial stage toward their application to improve SCFA deficiency.

Quantification of short-chain fatty acids in human stool samples by LC-MS/MS following derivatization with aniline analogues

The gut microbiome produces a range of short chain fatty acids (SCFA) crucially linked with diet and nutrition, metabolism, gastrointestinal health and homeostasis. SCFA are primarily measured using gas or liquid chromatography-mass spectrometry (LC/MS) after undergoing chemical derivatization. Here we assess the merits of a derivatization protocol using aniline and two aniline analogues (3-phenoxyaniline and 4-(benzyloxy)aniline) for the targeted LC-MS/MS quantification of nine SCFA (acetic, propionic, butyric, valeric, caproic acid, isobutyric, isovaleric, 2-methylbutyric, and 2-ethylbutyric acid). Evaluation of product ion spectra and optimization of MS detection conditions, provided superior detection sensitivity for 3-phenoxyaniline and 4-(benzyloxy)aniline compared to aniline. We developed a facile SCFA derivatization method using 3-phenoxyaniline under mild reaction conditions which allows for the simultaneous quantification of these SCFA in human stool samples in under eleven minutes using multiple reaction monitoring LC-MS/MS. The method was successfully validated and demonstrates intra- and inter-day accuracy (88.5–103% and 86.0–109%) and precision (CV of 0.55–7.00% and 0.33–9.55%) with recoveries (80.1–87.2% for LLOQ, 88.5–93.0% for ULOQ) and carry-over of (2.68–17.9%). Selectivity, stability and matrix effects were also assessed and satisfied validation criteria. Method applicability was demonstrated by analysing SCFA profiles in DNA-stabilized human stool samples from newly diagnosed colorectal cancer patients prior to surgery. The development of this improved method and its compatibility to measure SCFAs from DNA-stabilized stool will facilitate studies investigating the gut microbiome in health and disease.

High throughput FAMS – A fatty acid mass spectrometry method for monitoring polysorbate hydrolysis in QC

Surfactant degradation in biopharmaceuticals has recently gained significant attention in the pharmaceutical industry. Specifically, hydrolytic degradation of polysorbates, leading to the release of free fatty acids potentially forming visible particles, is a key theme in technical development. To address this emerging topic, we present the development of a fully automated liquid-chromatography single quad mass detector method for the quantification of free fatty acids in biopharmaceuticals. For the first time, we have quantified the longer chain fatty acid degradation products of polysorbate, palmitic and stearic acid, allowing reliable detection and early critical insights for process improvements. This high-throughput method was validated underlining its robust performance in an interlaboratory trial as well as high flexibility allowing different robotic platforms and preparation techniques. The combination of automated sample preparation, separation by liquid chromatography and single quad mass detection makes the validated fatty acid mass spectrometry assay ready for routine use in a regulated environment.

Different gut microbiota in U.S. formula-fed infants consuming a meat vs. dairy-based complementary foods: A randomized controlled trial.

This project aimed to evaluate the impact of meat- vs. dairy-based complementary foods on gut microbiota and whether it relates to growth. Full-term, formula-fed infants were recruited from the metro Denver area (Colorado, US) and randomized to a meat- or dairy-based complementary diet from 5 to 12 months of age. Infant's length and weight were measured, and stool samples were collected at 5, 10, and 12 months for 16S rRNA gene sequencing and short-chain fatty acids (SCFAs) quantification. Sixty-four infants completed the dietary intervention (n = 32/group). Weight-for-age Z (WAZ) scores increased in both groups and length-for-age Z scores (LAZ) increased in the meat group only, which led to a significant group-by-time interaction (P = 0.02) of weight-for-length Z (WLZ) score. Microbiota composition (Beta-diversity) differed between groups at 12 months (weighted PERMANOVA P = 0.01) and had a group-by-time interaction of P = 0.09. Microbial community richness (Chao1) increased in the meat group only. Genus Akkermansia had a significant group-by-time interaction and increased in the dairy group and decreased in the meat group. A significant fold change of butyric acid from 5 to 12 months was found in the meat group (+1.75, P = 0.011) but not in the dairy group. Regression analysis showed that Chao1 had a negative association with WLZ and WAZ. Several genera also had significant associations with all growth Z scores. Complementary feeding not only impacts infant growth but also affects gut microbiota maturation. Complementary food choices can affect both the gut microbiota diversity and structures and these changes in gut microbiota are associated with infant growth.

Identification and quantification of branched-chain fatty acids and odd-chain fatty acids of mammalian milk, dairy products, and vegetable oils using GC/TOF-MS

Branched-chain fatty acids (BCFAs) and odd-chain fatty acids (OCFAs) in mammalian milk, dairy products, and vegetable oils were investigated; 72 fatty acids, including 16 BCFAs and 10 OCFAs, were identified and quantified using GC/TOF-MS. Human milk showed the lowest content of BCFA (0.4% of total FA) and the content in other milk ranged from 2 to 6%, of which ai17:0, i17:0, ai15:0 and i15:0 were most abundant. Differences in the content of OCFA in mammalian milk trended similarly to differences in BCFA content. The molecular species of BCFA in cow milk-derived dairy products were identical, but the content was different, with higher content in fermented dairy products. Apparent differences in BCFA and OCFA composition were found in infant formulas, strongly related to their fat sources. Vegetable oils contained no BCFA and a small amount of OCFA. A scientific basis for further investigation of high-BCFA and OCFA dairy products is provided.