Search Results (12)

In France, sickle cell disease newborn screening (SCD NBS) has been targeted to at-risk regions since 1984, but generalization to the whole population will be implemented from November 2024. Although tandem mass spectrometry (MS/MS) is already used for the NBS of several inherited metabolic diseases, its application for SCD NBS has not been widely adopted worldwide. The aim of this study was to evaluate a dedicated MS/MS kit (Targeted MS/MS Hemo, ZenTech, LaCAR Company, Liege, Belgium) for SCD NBS and to compare the results obtained with those from an NBS reference center using matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) and cation-exchange high-performance liquid chromatography (CE-HPLC, Variant NBS, Biorad Laboratories, Inc., Hercules, CA, USA) as confirmatory method. The MS/MS Hemo kit was used according to the manufacturer’s instructions and performed on a Waters Xevo TQ-D (Waters Corporation, USA). The software provided by the manufacturer was used for the calculation and analysis of peptide signal ratios. Among the 1333 samples, the results of 1324 samples were consistent with the HPLC and/or MALDI-TOF results (1263 FA, 50 FAS, 7 FAC, 1 FAO-Arab, and 3 FS). All the discordant results (one FAS on MS/MS vs. FA in CE-HPLC, one FA on MS/MS vs. FAS in CE-HPLC, seven FS on MS/MS vs. FAS in CE-HPLC) were corrected after modifying the peptide signal ratios thresholds, allowing the MS/MS Hemo kit to achieve near-100% sensitivity and specificity for SCD NBS. In conclusion, the MS/MS Hemo kit appears to be an effective method for SCD NBS, particularly for laboratories already equipped with MS/MS technology. However, these results should be confirmed in a larger cohort including a greater number of positive samples for SCD. Full article

In the process of considering the documented health benefits of Viburnum opulus L. (V. opulus), including its anti-inflammatory and antioxidant activities, the present study was designed to qualitatively and quantitatively evaluate the biochemical profile and antimicrobial potency of four commercially available V. opulus extracts. These extracts were obtained from its flowers, bark, berries, and a mixture thereof by cold ultrasound-assisted extraction. An examination of the V. opulus extracts indicated a relative abundance of group compounds, such as phenolics, flavonoids, tannins, and anthocyanins, which are responsible for antioxidant activity (AOA). The widest range in all of the four group compounds was detected in the V. opulus extract sourced from berries, whereas the narrowest range was found in those obtained from flowers. The HPLC-ESI-TQ-MS/MS technique displayed relative fluctuations in the concentrations of individual amino acids (AAs) over the four V. opulus extracts. The prevalence of proline was marked in the flower-derived extract, which made up 63.3% of the total AAs, while aspartic and glutamic acids dominated in the berry-derived extract by contributing up to 29.2 and 24.4% to the total AA content, respectively. Profiling of the individual phenolic compounds disclosed the superiority of chlorogenic acid (up to 90.3%) in the berry and mixed extracts, as well as catechin (up to 57.7%) and neochlorogenic acid (11.1%) in the bark extract, which conveyed a remarkable contribution toward antimicrobial activity. The lowest content of individual phenolics was found in the flower extract. Owing to its substantially denser bioactive composition, the V. opulus berries and bark extracts exhibited markedly better AOA, which was pinpointed by three independent methods, i.e., DPPH^•, FRAP, and ABTS^•+, than those obtained from flowers or a mixture of V. opulus morphological parts. As part of the antimicrobial activity testing, the V. opulus extracts exhibited outstanding inhibitory activity and a homeopathic mode of action. The V. opulus extracts obtained from a mixture, bark, and berries were more active against 8 out of 19 selected test microorganisms at minimum inhibitory concentration (MIC) values that ranged from 0.24 to 0.49 µL mL⁻¹. Overall, the extracts of V. opulus were found to be effective against Gram-positive and Gram-negative bacteria. However, their conceivable exploitation as functional or pharmaceutical ingredients must be further clarified within in vivo models. Full article

The profile of amino acids and mono- and disaccharides in conventional polyfloral honey originated from Latvia and Tajikistan and less found in nature bumblebee honey from Russia was investigated. The analysis of free amino acids (FAAs) accomplished by multiple reaction monitoring (MRM) using triple quadrupole mass selective detection (HPLC-ESI-TQ-MS/MS) revealed the presence of 17 FAAs. The concentration of FAAs varied in the range of 0.02–44.41 mg 100 g⁻¹ FW. Proline was the main representative of FAAs, contributing to the total amount of FAAs from 41.7% to 80.52%. The highest concentration of proline was found in bumblebee and buckwheat honey, corresponding to 44.41 and 41.02 mg 100 g⁻¹, respectively. The concentration of essential amino acids (AAs), i.e., leucine, and isoleucine was found to be the highest in buckwheat honey contributing up to 12.5% to the total amount of FAAs. While, the concentration of branched-chain AAs fluctuated within the range of 1.08–31.13 mg 100 g⁻¹ FW, with buckwheat honey having the highest content and polyfloral honey the lowest, respectively. The results of this study confirmed the abundance of FAAs both in honeybee and bumblebee honey. However, the concentration of individual FAAs, such as proline, aspartic acid, leucine, and isoleucine in bumblebee honey was many folds higher than observed in honeybee polyfloral honey. Full article

The present work was undertaken to elucidate the potential contribution of biosynthetically produced ferulic acid (FA) via enzymatic hydrolysis (EH) of rye bran (RB) to the formation of silver nanoparticles (AgNPs) during green synthesis. An analytical approach accomplished by multiple reaction monitoring (MRM) using triple quadrupole mass selective detection (HPLC-ESI-TQ-MS/MS) of the obtained hydrolysate revealed a relative abundance of two isomeric forms of FA, i.e., trans-FA (t-FA) and trans-iso-FA (t-iso-FA). Further analysis utilizing high-performance liquid chromatography with refractive index (HPLC-RID) detection confirmed the effectiveness of RB EH, indicating the presence of cellulose and hemicellulose degradation products in the hydrolysate, i.e., xylose, arabinose, and glucose. The purification process by solid-phase extraction with styrene-divinylbenzene-based reversed-phase sorbent ensured up to 116.02 and 126.21 mg g⁻¹ of t-FA and t-iso-FA in the final eluate fraction, respectively. In the green synthesis of AgNPs using synthetic t-FA, the formation of NPs with an average size of 56.8 nm was confirmed by scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) techniques. The inclusion of polyvinylpyrrolidone (PVP-40) in the composition of NPs during synthesis favorably affected the morphological features, i.e., the size and shape of AgNPs, in which as big as 22.4 nm NPs were engineered. Meanwhile, nearly homogeneous round-shaped AgNPs with an average size of 16.5 nm were engineered using biosynthetically produced a mixture of t-FA and t-iso-FA and PVP-40 as a capping agent. The antimicrobial activity of AgNPs against Gram-positive and Gram-negative bacteria, including Pseudomonas aeruginosa, E. coli, Enterococcus faecalis, Bacillus subtilis, and Staphylococcus aureus was confirmed by the disk diffusion method and additionally supported by values of minimum inhibitory (MIC) and bactericidal (MBC) concentrations. Given the need to reduce problems of environmental pollution with cereal processing by-products, this study demonstrated a technological solution of RB rational use in the sustainable production of AgNPs during green synthesis. The AgNPs can be considered as active pharmaceutical ingredients (APIs) to be used for developing new antimicrobial agents and modifying therapies in treating multi-drug resistant (MDR) pathogens. Full article

A novel stability-indicating, reversed-phase, high-performance liquid chromatography (RP-HPLC) method was developed and validated for the determination of favipiravir in an oral suspension. The effective separation of favipiravir and its degradation products was achieved on a Zorbax Eclipse Plus C18 column (5 μm particle size, 150 mm length × 4.6 mm diameter). The mobile phase was prepared by mixing 5 mM of phosphate buffer (pH 3.5) and methanol in a 75:25 v/v ratio delivered at a 1.0 mL/min flow rate. The eluents were monitored using a photodiode array detector at a wavelength of 322 nm. The stability-indicating nature of this method was evaluated by performing force degradation studies under various stress conditions, such as acidic, alkali, oxidative, thermal, and photolytic degradation. Significant degradation was observed during the alkali stress degradation condition. The degradation products generated during various stress conditions were well separated from the favipiravir peak. In addition, the major degradation product formed under alkali stress conditions was identified using UPLC-ESI-TQ-MS/MS and NMR. Method validation was performed according to the ICH Q2 (R1) guideline requirements. The developed method is simple, accurate, robust, and reliable for routine quality control analysis of favipiravir oral suspensions. Full article

Horse chestnut (Aesculus hippocastanum) flower is a traditional medicine applied to alleviate symptoms of chronic venous insufficiency (CVI). However, its flavonoid-based composition has not been sufficiently recognized, and the data supporting its traditional application are lacking. In the work, 43 constituents were detected by UHPLC–PDA–ESI–TQ–MS/MS (flavonoids, phenolic acids, flavanols, and coumarins), including 31 reported in the flower for the first time. The quantitative HPLC–PDA study (developed and validated for quality control purposes) indicated the fractionated extraction as an efficient method for enhancing the total polyphenol content (TPHC) in the extracts (up to 414.06 mg/g) and kaempferol glycosides as their dominant constituents (75.05–82.14% TPHC). The activity studies showed significant scavenging properties of the extracts and their constituents towards reactive oxygen species (especially against highly reactive hydroxyl radical, with capacities up to 7.85 mmol ascorbic acid equivalents/g). Moreover, the analytes relevantly protected human plasma biomolecules from peroxynitrite-induced oxidative/nitrative damage; at 1–50 µg/mL, they hindered the protein nitration and lipid peroxidation, decreasing the levels of 3-nitrotyrosine (by up to 50%) and thiobarbituric acid reactive substances (by up to 70%), respectively. The extracts also averted the depletion of plasma thiols (by up to 67%) and improved the non-enzymatic antioxidant capacity of plasma. The demonstrated mechanisms might be partly responsible for the efficacy of the flower in CVI. Additionally, the anti-aggregatory and anticoagulant properties of the extracts were found only mild or negligible, which suggests that they may be safely applied with drugs impacting the coagulation process. Full article

Arbutus unedo L. berries have traditionally been used as edible and medicinal fruits in folk medicine for the treatment of some pathologies. Besides their good nutritional properties, these berries are rich in bioactive compounds. Accordingly, a HPLC-triple quadrupole (TQ)-MS/MS method was developed and validated for the first time to characterize the changes produced in the phenolic composition profile of A. unedo L. fruits through three ripening stages. Several hydroxycinnamic and hydroxybenzoic acids were detected, gallic acid being the highest phenolic acid quantified. Quercetin, quercetin-3-β-glucoside, rutin, and kaempferol were found in similar amounts at the different maturity stages. High amounts of (+)-catechin, procyanidin B2, epigallocatechin, and (−)-epigallocatechin gallate were observed in the unripe fruits. Naringin and vitexin were also quantified. Stilbenes, such as trans-resveratrol and trans-piceid, were identified for the first time in these fruits. Total phenolic compounds, total anthocyanin compounds, and radical scavenging activity were also determined in the fruits harvested at different years in several locations of the Iberian Peninsula at different ripening stages. The results confirmed that these fruits are an excellent source of these bioactive compounds (mainly flavanols) with high antioxidant activity, which could be used as a new source for preparation of nutraceutical or functional foods. Full article

Antimicrobials premixes are the presentation of choice to administer drugs simultaneously to groups of animals in intensive husbandry systems that require treatment for pathologies of bacterial origin. Among the premixes available for use in poultry, florfenicol and oxytetracycline are commonly administered via food or water. However, their actual concentration in premixes must meet on-label statements to ensure plasma concentrations reach effective therapeutic levels. Hence, this work was designed for the purpose of verifying whether the concentration of antimicrobial present in five premixes matched their on-label statement. Three oxytetracycline premixes, and two of florfenicol, were analysed using a Xevo TQ-S micro UPLC-MS/MS, and an ABSciex API4000 HPLC-MS/MS, respectively. Analytical methodologies were implemented and validated, showing an R² ≥ 0.99 for the calibration curves. Oxytetracycline was detected in these premixes at concentrations exceeding on-label statements by 13.28%, 21.54%, and 29.68%, whereas florfenicol concentrations detected in premixes were 13.06% and 14.75% lower than expected. Consequently, this work shows that the concentration of active ingredients that are present in commercial formulations effectively differ from those stated on premix labels, and it also highlights how unpredictable their range of variability might be. This must be addressed through solid and updated laws that guarantee an effective pharmaceutical product. Full article

Uncaria tomentosa, which is widely commercialized as an herbal medicine, constitutes an important source of secondary metabolites with diverse biological activities. For instance, we have previously reported, for the first time, of a polyphenolic profile rich in proanthocyanidins from extracts of U. tomentosa plants, as well as their antioxidant capacity, antimicrobial activity on aerial bacteria, and cytotoxicity on cancer cell lines. These promising results prompted this research to evaluate the polyphenolic contents of U. tomentosa commercial products. We report a detailed study on the polyphenolic composition of extracts from U. tomentosa bark products (n = 18) commercialized in Costa Rica and Spain. Using HPLC-DAD/TQ-ESI-MS, a total of 25 polyphenolic compounds were identified, including hydroxybenzoic and hydroxycinnamic acids, flavan-3-ol monomers, procyanidin dimers, procyanidin trimers, as well as propelargonidin dimers. Our findings on the polyphenolic profile for all commercial samples show analogous composition to previous reports on U. tomentosa bark material, for instance a 41–49% content of procyanidin dimers and the presence of propelargonidin dimers (8–15%). However, most of the 18 commercial samples exhibit low proanthocyanidin contents (254.8–602.8 µg/g), more similar to previous U. tomentosa inner bark reports, while some exhibit better results, with one sample (SP-2) showing the highest contents (2386.5 µg/g) representing twice the average value of all 18 commercial products. This sample also exhibits the highest total phenolics (TP) and total proanthocyanidins (PRO) contents, as well as the highest Oxygen Radical Absorbance Capacity (ORAC) value (1.31 µg TE/g). One-way Analysis of Variance (ANOVA) with a Tukey post hoc test indicated significant difference (p < 0.05) between products from Costa Rica and Spain for TP and PRO findings, with samples from Spain exhibiting a higher average value. In addition, Pearson correlation analysis results showed a positive correlation (p < 0.05) between TP, PRO, and ORAC results, and an especially important correlation between ORAC antioxidant values and procyanidin dimers (r = 0.843, p < 0.05), procyanidin trimers (r = 0.847, p < 0.05), and propelargonidin dimers (r = 0.851, p < 0.05) contents. Finally, Principal Component Analysis (PCA) results indicated some variability in the composition regardless of their origin. However, only one sample (SP-2) stands out significatively, showing the highest PC1 because of its particularly high proanthocyanidins contents, which could be attributed to the 15% bark polyphenolic extract labeled in this commercial product, which differentiate this sample from all other 17 commercial samples. Therefore, our findings confirmed previous results on the value of extracts in the elaboration of potential commercial products from U. tomentosa, rich in proanthocyanidins and exhibiting high antioxidant activity. Full article

Uncaria tomentosa constitutes an important source of secondary metabolites with diverse biological activities mainly attributed until recently to alkaloids and triterpenes. We have previously reported for the first-time the polyphenolic profile of extracts from U. tomentosa, using a multi-step process involving organic solvents, as well as their antioxidant capacity, antimicrobial activity on aerial bacteria, and cytotoxicity on cancer cell lines. These promising results prompted the present study using food grade solvents suitable for the elaboration of commercial extracts. We report a detailed study on the polyphenolic composition of aqueous and ethanolic extracts of U. tomentosa bark and leaves (n = 16), using High Performance Liquid Chromatography coupled with Mass Spectrometry (HPLC-DAD/TQ-ESI-MS). A total of 32 compounds were identified, including hydroxybenzoic and hydroxycinnamic acids, flavan-3-ols monomers, procyanidin dimers and trimers, flavalignans–cinchonains and propelargonidin dimers. Our findings showed that the leaves were the richest source of total phenolics and proanthocyanidins, in particular propelargonidin dimers. Two-way Analysis of Variance (ANOVA) indicated that the contents of procyanidin and propelargonidin dimers were significantly different (p < 0.05) in function of the plant part, and leaves extracts showed higher contents. Oxygen Radical Absorbance Capacity (ORAC) and 2,2-diphenyl-1-picrylhidrazyl (DPPH) values indicated higher antioxidant capacity for the leaves (p < 0.05). Further, correlation between both methods and procyanidin dimers was found, particularly between ORAC and propelargonidin dimers. Finally, Principal Component Analysis (PCA) analysis results clearly indicated that the leaves are the richest plant part in proanthocyanidins and a very homogenous material, regardless of their origin. Therefore, our findings revealed that both ethanol and water extraction processes are adequate for the elaboration of potential commercial extracts from U. tomentosa leaves rich in proanthocyanidins and exhibiting high antioxidant activity. Full article

Salvia miltiorrhiza is a traditional Chinese herbal medicine containing multiple components that contribute to its notable bioactivities. This article investigated the distribution and dynamic changes of chemical constituents in various parts of S. miltiorrhiza from different growth periods. An ultra-high performance liquid chromatography-triple quadrupole mass spectrometer (UPLC-TQ-MS/MS) and high-performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) methods were developed for accurate determination of 24 compounds (including phenolic acids, flavonoids, triterpenes, and saccharides) in S. miltiorrhiza. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. Results indicated that there were category and quantity discrepancies in different parts of the plant, for the roots mainly contained salvianolic acids and tanshinones, and most of the saccharides are stachyose. In the aerial parts, salvianolic acids, flavonoids, and triterpenes, except the tanshinones, were detected, and the saccharides were mainly monosaccharides. Dynamic accumulation analysis suggested the proper harvest time for S. miltiorrhiza Bunge was the seedling stage in spring, and for the aerial parts was July to August. This study provided valuable information for the development and utilization value of the aerial parts of S. miltiorrhiza and was useful for determining the optimal harvest time of the plant. Full article

Detection of sulfur-fumigated Paeoniae Alba Radix (PAR) in different complex preparations is challenging due to the relatively lower content of PAR and interference from more complicated components in complex preparations with different multiple constituent herbs. In this study, a high performance liquid chromatography- triple-quadrupole tandem mass spectrometry method was developed for detecting sulfur-fumigated PAR in different complex preparations. Paeoniflorin, the major component of PAR, and paeoniflorin sulfonate, the characteristic artifact transformed from paeoniflorin during sulfur-fumigation of PAR, were used as chemical markers. Multiple reaction monitoring (MRM) scan was employed to maximize sensitivity and selectivity. Through optimizing full mass scan and daughter ion scan conditions, two mass transitions were selected and employed respectively for unequivocal identification of paeoniflorin and paeoniflorin sulfonate. The detection limits for paeoniflorin and paeoniflorin sulfonate using MRM were much lower than those detected with UV 270 nm. Paeoniflorin and paeoniflorin sulfonate could be simultaneously detected in different commercial PAR-containing complex preparations without interference of other components using the established method, indicating that the newly established method was selective and sensitive enough for screening sulfur-fumigated PAR in commercial complex preparations. Full article