UHPLC-PAD Protocol for the Simultaneous Identification of Polyphenols and Bitter Acids: Organoleptic and Nutraceutical Fingerprinting of Bergamot-Flavored Craft Beer.

In this study, a UHPLC-PDA method for the simultaneous identification of polyphenols and bitter acids (alpha, beta, and isoalpha) in beer was developed. The resulting chemical profiles were leveraged to distinguish the characteristics of four (IPA, Lager, Blanche, ALE) bergamot-flavored beers, produced on a pilot-scale plant. In a streamlined 29 min analysis, thirty polyphenols and fourteen bitter acids were successfully identified under optimized separation conditions. Validation, encompassing parameters such as LOD (from 0.028 ppm for isorhamnetin to 0.106 for narirutin), LOQ (from 0.077 ppm for naringenin to 0.355 for narirutin), R2 (always more than 0.9992), repeatability (from 0.67% for tangeretin to 6.38% for myricetin), and reproducibility (from 0.99% for sinensetin to 6% for naringin), was conducted for polyphenol quantification using constructed calibration curves with seven levels. Exploring polyphenolic components as potential discriminators among different beer styles, a total of thirty-two polyphenolic compounds were identified and quantified, including characteristic bergamot peel polyphenols like neoeriocitrin (from 7.85 ppm for CBS2 to 11.95 ppm in CBS1); naringin (from 4.56 ppm for CBS4 to 10.96 in CBS1), and neohesperidin (from 5.93 in CBS3 to 15.95 for CBS2). The multivariate analysis provided additional insights into variations among specific beer styles, revealing discrepancies in the presence or relative concentrations of specific compounds linked to brewing ingredients and processes. This research enhances the fingerprinting of the chemistry governing beer quality through a straightforward and cost-effective analytical approach.

On-line coupling of supercritical fluid extraction with enantioselective supercritical fluid chromatography-triple quadrupole mass spectrometry for the determination of chiral pesticides in hemp seeds: A proof-of-principle study.

The present research can be considered as a proof-of-principle study focused on the determination of chiral pesticides using a supercritical fluid extraction instrument coupled on-line with an enantioselective supercritical fluid chromatography-triple quadrupole mass spectrometry. To the best of Authors' knowledge, this is the first description of an on-line approach for the extraction and determination of chiral pesticides. Metalaxyl, benalaxyl and dimethenamid were investigated in nine hemp seed samples belonging to four varieties of Cannabis sativa; only in one case a pesticide was found at levels above the method limit of quantification (LoQ), though within the EU maximum residue level value. The figures-of-merit determined were linearity, precision, limit of detection (LoD), and LoQ. Regression coefficients were between 0.9856 and 0.9973, the LoDs were in the 0.04-0.41 µg kg-1 range, the LoQs were in the 0.12-1.38 µg kg-1 range, while coefficients of variation were between 1 and 3% (10 µg kg-1 level).

Interlaboratory study of a supercritical fluid chromatography method for the determination of pharmaceutical impurities: Evaluation of multi-systems reproducibility.

Modern supercritical fluid chromatography (SFC) is now a well-established technique, especially in the field of pharmaceutical analysis. We recently demonstrated the transferability and the reproducibility of a SFC-UV method for pharmaceutical impurities by means of an inter-laboratory study. However, as this study involved only one brand of SFC instrumentation (Waters®), the present study extends the purpose to multi-instrumentation evaluation. Specifically, three instrument types, namely Agilent®, Shimadzu®, and Waters®, were included through 21 laboratories (n = 7 for each instrument). First, method transfer was performed to assess the separation quality and to set up the specific instrument parameters of Agilent® and Shimadzu® instruments. Second, the inter-laboratory study was performed following a protocol defined by the sending lab. Analytical results were examined regarding consistencies within- and between-laboratories criteria. Afterwards, the method reproducibility was estimated taking into account variances in replicates, between-days and between-laboratories. Reproducibility variance was larger than that observed during the first study involving only one single type of instrumentation. Indeed, we clearly observed an 'instrument type' effect. Moreover, the reproducibility variance was larger when considering all instruments than each type separately which can be attributed to the variability induced by the instrument configuration. Nevertheless, repeatability and reproducibility variances were found to be similar than those described for LC methods; i.e. reproducibility as %RSD was around 15 %. These results highlighted the robustness and the power of modern analytical SFC technologies to deliver accurate results for pharmaceutical quality control analysis.

Apocarotenoids profiling in different Capsicum species.

The present study report on the application of an on line supercritical fluid extraction-supercritical fluid chromatography-triple quadrupole/mass spectrometry methodology to the first apocarotenoids profiling of seventeen different chilli peppers cultivars belonging to Capsicum annuum, Capsicum baccatum and Capsicum chinense species. A total of 19 free apocarotenoids and 8 apocarotenoids fatty acid esters were identified; ß-Apo-8'-carotenal and Apo-8'-zeaxanthinal were also quantified and the ß-Apo-8'-carotenal occurrence was in the percentage ranges relative to ß-carotene of 0.03-3.87%. PCA was performed as a multivariate display method on the quantified carotenoids and apocarotenoids, in order to visualize the data structure. Moreover, different ε-apoluteinals and 4-oxo-apo-ß-carotenals were detected in Capsicum species also for the first time and, to the best of authors knowledge, in any food matrix.

Characterization of Phenolic Compounds, Vitamin E and Fatty Acids from Monovarietal Virgin Olive Oils of "Picholine marocaine" Cultivar.

Olive oil is an important product in the Mediterranean diet, due to its health benefits and sensorial characteristics. Picholine marocaine is the most cultivated variety in Morocco. The present research aims to evaluate the phenolic compounds, vitamin E and fatty acids of commercial Picholine marocaine virgin olive oils (VOOs) from five different North Moroccan provinces (Chefchaouen, Taounate, Errachidia, Beni Mellal and Taza), using HPLC-photodiode array (PDA)/electrospray ionization (ESI)-MS, normal phase (NP)-HPLC/ fluorescence detector (FLD) and GC-flame ionization detector (FID)/MS, respectively. The obtained results showed an average content of 130.0 mg kg-1 of secoiridoids (oleuropein aglycone, 10-hydroxy-oleuropein aglycone and ligstroside aglycone, oleocanthal and oleacein), 108.1 mg kg-1 of phenolic alcohols (tyrosol and hydroxytyrosol), 34.7 mg kg-1 of phenolic acids (caffeic acid, ferulic acid and elenolic acid), and 8.24 mg kg-1 of flavonoids (luteolin, luteolin glucoside, apigenin). With regard to vitamin E, α-tocopherol was the most abundant vitamin E (57.9 mg kg-1), followed by α-tocotrienol (2.5 mg kg-1), γ-tocopherol (4.5 mg kg-1) and ß-tocopherol (1.9 mg kg-1), while δ-tocopherol was not detected. Moreover, 14 fatty acids were found and, among them, oleic acid (76.1%), linoleic acid (8.1%) palmitic acid (8.7%) and stearic acid (2.5%) were the major fatty acids detected. Finally, heat map and principal component analysis allowed us to classify the studied provinces in terms of VOO chemical composition: Chefchaouen (tyrosol and hydroxytyrosol), Taounate (oleuropein aglycone), Errachidia (ferulic acid, w-3 and w-6), Beni Mellal (oleocanthal) and Taza (luteolin and oleic acid).

Concentration of Potentially Bioactive Compounds in Italian Extra Virgin Olive Oils from Various Sources by Using LC-MS and Multivariate Data Analysis.

High quality extra virgin olive oils represent an optimal source of nutraceuticals. The European Union (EU) is the world's leading olive oil producer, with the Mediterranean region as the main contributor. This makes the EU the greatest exporter and consumer of olive oil in the world. However, small olive oil producers also contribute to olive oil production. Beneficial effects on human health of extra virgin olive oil are well known, and these can be correlated to the presence of vitamin E and phenols. Together with the origin of the olives, extraction technology can influence the chemical composition of extra virgin olive oil. The aim of this study was to investigate the concentration of potentially bioactive compounds in Italian extra virgin olive oils from various sources. For this purpose, vitamin E and phenolic fractions were characterized using high-performance liquid chromatography (HPLC) coupled with fluorescence, photodiode array and mass spectrometry detection in fifty samples of oil pressed at industrial plants and sixty-six samples of oil produced in low-scale mills. Multivariate statistical data analysis was used to determine the applicability of selected phenolic compounds as potential quality indicators of extra virgin olive oils.

Determination of free apocarotenoids and apocarotenoid esters in human colostrum.

The presence of carotenoids in human colostrum has been reported in the literature, and xanthophyll esters in human colostrum were recently detected for the first time. However, no published studies have reported whether apocarotenoids, which are metabolites derived from carotenoid enzymatic or nonenzymatic oxidative cleavage, are present in human colostrum. Therefore, the purpose of the present study was to search for the possible occurrence of apocarotenoids, including apocarotenoid esters, in human colostrum for the first time by applying an online supercritical fluid extraction-supercritical fluid chromatography-tandem mass spectrometry methodology. Recent evidence related to apocarotenoid transcriptional activity has suggested that they may have beneficial health properties superior to those of their parent carotenoids. Three different apocarotenoids, namely apo-8'-ß-carotenal, apo-8'-lycopenal, and ß-citraurin, were identified in intact human colostrum samples, with average concentrations of 85 nmol L-1, 54.6 nmol L-1, and 75.4 nmol L-1, respectively. The overall detection of 16 different free apocarotenoids and 10 different apocarotenoid fatty acid esters in human colostrum was achieved here for the first time. Their occurrence in human colostrum certainly has implications for newborn health status, since colostrum is the only form of food for the newborn during the very first days of life. Graphical abstract.

Green Extraction Approaches for Carotenoids and Esters: Characterization of Native Composition from Orange Peel.

Orange peel is a by-product produced in large amounts that acts as a source of natural pigments such as carotenoids. Xanthophylls, the main carotenoid class found in citrus fruit, can be present in its free form or esterified with fatty acids, forming esters. This esterification modifies the compound's chemical properties, affecting their bioavailability in the human body, and making it important to characterize the native carotenoid composition of food matrices. We aimed to evaluate the non-saponified carotenoid extracts of orange peel (cv. Pera) obtained using alternative green approaches: extraction with ionic liquid (IL), analyzed by high performance liquid chromatography coupled to a diode array detector with atmospheric pressure chemical ionization and mass spectrometry HPLC-DAD-APCI-MS, and supercritical fluid extraction (SFE), followed by supercritical fluid chromatography with atmospheric pressure chemical ionization and triple quadrupole mass spectrometry detection (SFC-APCI/QqQ/MS) in an online system. Both alternative green methods were successfully applied, allowing the total identification of five free carotenoids, one apocarotenoid, seven monoesters, and 11 diesters in the extract obtained with IL and analyzed by HPLC-DAD-APCI-MS, and nine free carotenoids, six carotenoids esters, 19 apocarotenoids, and eight apo-esters with the SFE-SFC-APCI/QqQ/MS approach, including several free apocarotenoids and apocarotenoid esters identified for the first time in oranges, and particularly in the Pera variety, which could be used as a fruit authenticity parameter.

First Apocarotenoids Profiling of Four Microalgae Strains.

Both enzymatic or oxidative carotenoids cleavages can often occur in nature and produce a wide range of bioactive apocarotenoids. Considering that no detailed information is available in the literature regarding the occurrence of apocarotenoids in microalgae species, the aim of this study was to study the extraction and characterization of apocarotenoids in four different microalgae strains: Chlamydomonas sp. CCMP 2294, Tetraselmis chuii SAG 8-6, Nannochloropsis gaditana CCMP 526, and Chlorella sorokiniana NIVA-CHL 176. This was done for the first time using an online method coupling supercritical fluid extraction and supercritical fluid chromatography tandem mass spectrometry. A total of 29 different apocarotenoids, including various apocarotenoid fatty acid esters, were detected: apo-12'-zeaxanthinal, ß-apo-12'-carotenal, apo-12-luteinal, and apo-12'-violaxanthal. These were detected in all the investigated strains together with the two apocarotenoid esters, apo-10'-zeaxanthinal-C4:0 and apo-8'-zeaxanthinal-C8:0. The overall extraction and detection time for the apocarotenoids was less than 10 min, including apocarotenoids esters, with an overall analysis time of less than 20 min. Moreover, preliminary quantitative data showed that the ß-apo-8'-carotenal content was around 0.8% and 2.4% of the parent carotenoid, in the C. sorokiniana and T. chuii strains, respectively. This methodology could be applied as a selective and efficient method for the apocarotenoids detection.

Carotenoids and apocarotenoids determination in intact human blood samples by online supercritical fluid extraction-supercritical fluid chromatography-tandem mass spectrometry.

A direct on-line method based on the coupling of supercritical fluid extraction and supercritical fluid chromatography with triple quadrupole mass spectrometry detection (SFE-SFC-QqQ/MS) for selected carotenoids determination and apocarotenoids detection in intact human blood was developed for the first time. Carotenoids and apocarotenoids were identified by using the available standard together with full scan, selected ion monitoring (SIM), and multiple reaction monitoring (MRM) experiments. Moreover, ß-Cryptoxanthin, Zeaxanthin, ß-Carotene and Capsanthin were directly quantified by the developed methodology, using a multiple reaction monitoring (MRM) approach; the determined average content of ß-carotene was 123.8 nmol L-1 (range 18.7-485.1 nmol L-1), of ß-cryptoxanthin was 385.3 nmol L-1 (range 72.5-1920.3 nmol L-1), of zeaxanthin was 396.9 nmol L-1 (range < LoD - 1795.8 nmol L-1) and of capsanthin was 38.9 nmol L-1 (range < LoD - 188.4 nmol L-1). Analyses were carried out on 10 µL aliquots of intact blood samples without any preliminary treatment; the online extraction and chromatographic separation time was just over 20 min. The method was validated in terms of linearity, precision, limits of detection and quantification, and accuracy. Interestingly, ß-apo-12'-carotenal, apo-10'-zeaxanthinal, apo-12'-zeaxanthinal, apo-14'-zeaxanthinal, ε-apo-8-luteinal, ε-apo-12-luteinal and ε-apo-14-luteinal were detected in human blood, together with two zeaxanthin fatty acid esters, for the first time.