Folates play an important role in the human body and a deficiency of this vitamin can cause several diseases. Therefore, a reliable analytical method is crucial for the determination of folate vitamers in strawberries and other dietary folate sources. A stable isotope dilution LC-MS/MS method for analyzing folates in food was developed and validated. The folate vitamers Pteroylmonoglutamic acid, tetrahydrofolate, 5-methyltetrahydrofolate, and 5-formyltetrahydrofolate were quantified using 13C-labeled internal standards. Validation of the assay was accomplished by determining linearity, precision, recovery, limit of detection, and limit of quantification and revealed to be a precise, sensitive, and accurate method to determine folate vitamers. Strawberries are worldwide consumed and known to be a good dietary source of nutritive compounds. Using this method, folate concentrations in selected commercial strawberry cultivars and experimental breeding lines grown in Germany and Australia were investigated. Total folates varied from 59 to 153 μg/100 g on fresh weight basis. Furthermore, folate content after lyophilizing or freezing did not show any significant differences compared to fresh strawberries. However, significant losses of total folates in pureed strawberries could be observed after 5 days of storage with only 16% of the original concentration retained. In summary, some of the investigated strawberry cultivars/breeding lines can be considered as rich dietary sources of natural folates.

This study evaluates the potential mechanism of action and bioactivity of black tea and black tea pomace for type 2 diabetes prevention via inhibition of carbohydrate hydrolyzing enzymes. Black tea leaves were extracted in hot water and black tea pomace was extracted in 70% acetone. The phenolic content of the water extract (WBT) and pomace acetone extracts (AOBT) were 5.77 and 8.9 mg/mL, respectively, both based on the same concentration of solid tea in the extract. The water extract was subjected to C18 extraction and the resulting hydrophobic fraction (HBBT) was further subjected to LH-20 extraction to recover a low molecular weight phenolic enriched fraction (LMW) and a high molecular weight enriched fraction (HMW). The phenolic content of the LMW and HMW fraction were 1.42 and 2.66 mg/mL, respectively. Among water extracts the HMW fraction was most bioactive against α-glucosidase (IC50 = 8.97 μg/mL) followed by HBBT fraction (IC50 = 14.83 μg/mL). However, the HBBT fraction was the most bioactive fraction against α-amylase (IC50 = 0.049 mg/mL). The black tea pomace (AOBT) had significant α-glucosidase inhibitory activity (IC50 = 14.72 μg/mL) but lower α-amylase inhibitory activity (IC50 = 0.21 mg/mL). The phenolic profiles for LMW and HMW fractions were evaluated using HPLC and the differences between the two profiles were identified. Further research is underway to identify and evaluate the phenolic compounds that are present in the HMW fraction. Our findings suggest that black tea and black tea pomace has potential for carbohydrate hydrolyzing enzyme inhibition and this activity depends on high molecular weight phenolic compounds.

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As the popularity of tropical fruits has been increasing consistently during the last few decades, nutritional and health-related data about these fruits have been gaining more and more interest. Therefore, we analyzed 35 samples of tropical fruits and vegetables with respect to folate content and vitamer distribution in this study. The fruits and vegetables were selected by their availability in German supermarkets and were grouped according to their plant family. All fruits and vegetables were lyophilized and analyzed by stable isotope dilution assay (SIDA) and liquid chromatography mass spectrometry (LC-MS/MS). The results vary from 7.82 ± 0.17 µg/100 g in the horned melon to 271 ± 3.64 µg/100 g in the yellow passion fruit. The yellow passion fruit is a good source for meeting the recommended requirements, as just 110 g are needed to cover the recommended daily intake of 300 µg folate for adults; however, longan fruits, okras, pete beans, papayas, mangos, jack fruits, and feijoas are also good sources of folates. In conclusion, the study gives a good overview of the total folate content in a broad range of tropical fruits and vegetables and shows that some of these fruits definitely have the potential to improve the supply of this critical vitamin.

A multitude of human nutritional supplements based on Chlorella vulgaris biomass has recently been introduced to the specialty food market. In this study, an analysis of total folate contents in Chlorella sp. and a series of marine microalgae was conducted to evaluate folate content in alternative algae-based food production strains. For the first time, total folate content and vitamer distribution in microalgae were analyzed by stable isotope dilution assay (SIDA) using LC-MS/MS, which has demonstrated its superiority with respect to folate quantification. Consistently, high folate contents were detected in all examined microalgae samples. High folate concentrations of 3,460 ± 134 µg/100 g dry biomass were detected in freshly cultivated Chlorella vulgaris, notably also in other well-researched microalgae strains. To that end, the highest folate content currently documented for any algae sample was measured in the marine microalgae Picochlorum sp. isolate with values of 6,470 ± 167 µg/100 g dry biomass. This calls for alternative products based on other algae biomass. Our data indicate that freshwater and marine microalgae provide extremely high concentrations of folates, which warrant further studies on the regulation of pteroylpolyglutamates in algae as well as on bioaccessibility, absorption, and retention in humans.

Folates are a group of B9 vitamins playing an important role in many metabolic processes such as methylation reactions, nucleotide synthesis or oxidation and reduction processes. However, humans are not able to synthesize folates de novo and thus rely on external sources thereof. Baker’s yeast (Saccharomyces cerevisiae) has been shown to produce high amounts of this vitamin but extensive identification of its folate metabolism is still lacking. Therefore, we optimized and compared different sample preparation and purification procedures applying solid phase extraction (SPE). Strong anion exchange (SAX), C18 and hydrophilic–lipophilic-balanced (HLB) materials were tested for their applicability in future metabolomics studies. SAX turned out to be the preferred material for the quantitative purification of folates. Qualification of several folate vitamers was achieved by ultra-high pressure liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-Q-ToF-MS) measurements and quantification was performed by liquid chromatography tandem mass spectrometry (LC-MS/MS) applying stable isotope dilution assays (SIDAs). The oxidation product s-pyrazino-triazine (MeFox) was included into the SIDA method for total folate determination and validation. Applying the best protocol (SAX) in regard to folate recovery, we analyzed 32 different vitamers in different polyglutamate states up to nonaglutamates, of which we could further identify 26 vitamers based on tandem-MS (MS2) spectra. Total folate quantification revealed differences in formyl folate contents depending on the cartridge chemistry used for purification. These are supposedly a result of interconversion reactions occurring during sample preparation due to variation in pH adjustments for the different purification protocols. The occurrence of interconversion and oxidation reactions should be taken into consideration in sample preparation procedures for metabolomics analyses with a focus on folates.

Spent yeast from beer manufacturing is a cost-effective and nutrient-rich starting material for the production of yeast extracts. In this study, it is shown how physiologically important ingredients in a yeast extract are influenced by the composition of the spent yeast from the brewing process. In pilot fermentations, the time of cropping (primary fermentation, lagering) of the spent yeast and the original gravity (12 • P, 16 • P, 20 • P) of the fermentation medium was varied, and four alternative non-Saccharomyces yeast strains were compared with two commercial Saccharomyces yeast strains. In addition, spent yeast was contaminated with the beer spoiler Lactobacillus brevis. The general nutrient composition (total protein, fat, ash) was investigated as well as the proteinogenic amino acid spectrum, the various folate vitamers (5-CH 3 -H 4 folate, 5-CHO-H 4 folate, 10-CHO-PteGlu, H 4 folate, PteGlu) and the biological activity (reduction, antioxidative potential) of a mechanically (ultrasonic sonotrode) and an autolytically produced yeast extract. All the investigated ingredients from the yeast extract were influenced by the composition of the spent yeast from the brewing process. The biodiversity of the spent yeast from the brewing process therefore directly affects the content of physiologically valuable ingredients of a yeast extract and should be taken into consideration in industrial manufacturing processes. cell that remains once the cell wall has been destroyed and removed [4][5][6]. The variety of different physiologically valuable substances in yeast cells offers the possibility of using them as yeast extract in different areas of the food industry [3,7]. As "yeast food", these extracts can therefore increase the free α-amino nitrogen (FAN) when fermenting beer worts with a high content of unmalted grains [8] or a high extract content (high-gravity worts) [9,10], and consequently improve the yeast's nutrient supply and fermentation performance [11,12]. Free proteinogenic amino acids supply the majority of the FAN [12]. The quantity and composition of the relevant amino acids are ultimately critical to performance during fermentation [13] and also impact the beer's aroma profile [14]. From a nutritional standpoint, yeast extracts from spent yeast supply a high concentration of essential and semi-essential amino acids for human beings [5,7]. Yeast extracts are also a good source of B vitamins [6,15]. Among these, the various naturally occurring folate vitamers play an essential role in the human diet, with the biologically active form 5-methyltetrahydrofolate (5-CH 3 -H 4 folate) fulfilling key metabolic tasks in human cells [16]. The bioactivity of yeast extracts, which is demonstrated in the form of reduction and anti-oxidative potential, also makes these extracts particularly interesting for the food industry [6,7,15,17].The majority of globally produced beer is manufactured by fermenting high-gravity worts [18]. The extract content of the wort is increased by adding sugar syrup, which modifies the nutr...