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Indacaterol, a β2 agonist prescribed for long-term management of patients with chronic obstructive pulmonary disease and asthma. In this study the first MISPE cartridges was developed using indacaterol as a template for its selective extraction from rat lung tissues, enabling precise pharmacokinetic evaluation at the drug's site of action. A molecular imprinting polymer was synthesized using indacaterol as a template, methacrylic acid as a functional monomer and ethylene glycol dimethacrylate as a cross-linker with a molar ratio (1: 4: 20). The polymer was characterized by a high binding capacity of 9840 ± 0.86 and high selectivity with an imprinting factor of 4.53 ± 0.12. The synthesized polymer was utilized as a sorbent in solid-phase extraction to purify and extract indacaterol from lung tissue matrix. The optimum molecularly imprinted solid-phase extraction (MISPE) conditions were 20.0 mg of molecular imprinting polymer and non-imprinting polymer, acetonitrile as the loading solvent, acetonitrile: water (20: 80; by volume) as the washing solvent, and methanol: acetic acid (90: 10; by volume) as the eluting solvent. A pharmacokinetic study was performed for indacaterol in rat lungs using the synthesized and optimized MISPE cartridge as a tool for sample purification. These parameters were determined in the lung tissues of rats emphasizing the local exposure of indacaterol to its target organ. The Cmax and Tmax were 51.020 ± 2.810 µg mL− 1 and 0.083 ± 0.001 h, respectively. The AUC 0−24 and AUC0 − inf were 175.920 ± 1.053 and 542.000 ± 5.245 µg h mL− 1, respectively. The elimination rate constant was 0.014 ± 0.00012 h− 1 and the half-life time was 48.510 ± 0.012 h. This study successfully developed and optimized MISPE cartridges using indacaterol as a template, enabling precise pharmacokinetic evaluation in rat lung tissues. The cartridges demonstrated high binding capacity and selectivity, providing crucial insights into the local exposure of indacaterol at its site of action. [ABSTRACT FROM AUTHOR]

Abstract Indacaterol, a β2 agonist prescribed for long-term management of patients with chronic obstructive pulmonary disease and asthma. In this study the first MISPE cartridges was developed using indacaterol as a template for its selective extraction from rat lung tissues, enabling precise pharmacokinetic evaluation at the drug’s site of action. A molecular imprinting polymer was synthesized using indacaterol as a template, methacrylic acid as a functional monomer and ethylene glycol dimethacrylate as a cross-linker with a molar ratio (1: 4: 20). The polymer was characterized by a high binding capacity of 9840 ± 0.86 and high selectivity with an imprinting factor of 4.53 ± 0.12. The synthesized polymer was utilized as a sorbent in solid-phase extraction to purify and extract indacaterol from lung tissue matrix. The optimum molecularly imprinted solid-phase extraction (MISPE) conditions were 20.0 mg of molecular imprinting polymer and non-imprinting polymer, acetonitrile as the loading solvent, acetonitrile: water (20: 80; by volume) as the washing solvent, and methanol: acetic acid (90: 10; by volume) as the eluting solvent. A pharmacokinetic study was performed for indacaterol in rat lungs using the synthesized and optimized MISPE cartridge as a tool for sample purification. These parameters were determined in the lung tissues of rats emphasizing the local exposure of indacaterol to its target organ. The Cmax and Tmax were 51.020 ± 2.810 µg mL− 1 and 0.083 ± 0.001 h, respectively. The AUC 0−24 and AUC0 − inf were 175.920 ± 1.053 and 542.000 ± 5.245 µg h mL− 1, respectively. The elimination rate constant was 0.014 ± 0.00012 h− 1 and the half-life time was 48.510 ± 0.012 h. This study successfully developed and optimized MISPE cartridges using indacaterol as a template, enabling precise pharmacokinetic evaluation in rat lung tissues. The cartridges demonstrated high binding capacity and selectivity, providing crucial insights into the local exposure of indacaterol at its site of action.

Relevance. Rhodiola rosea L. is a perennial herbaceous plant from the Crassulaceae family. The roots and rhizomes of R. rosea contain a range of biologically active compounds. This species is included in the State Pharmacopoeia of the Russian Federation. Medicinal preparations derived from them are widely used as adaptogens, general tonic and anti-stress agents. While the roots have been extensively studied, less attention has been given to the biologically active compounds found in the leaves of R. rosea. An introduced population of R. rosea grows on the territory of the botanical garden of medicinal plants of the All-Russian scientific research Institute of medicinal and aromatic plants (VILAR). The possibility of cultivating this species in the conditions of the Moscow region is being studied.Material and Methodology. The objective of this study was to investigate the composition of phenolic compounds in R. rosea leaves from VILAR biocollection using ultra-efficient liquid chromatography combined with mass spectrometry. Extraction was carried out with 80% acetone. An ultra-high performance liquid chromatographic system with a photodiode array detector and mass spectrometer was used for the analysis of phenolic compounds. Identification was carried out using UV- and mass spectrometry data.Results. As a result of UPLC-DD-MS analysis, peaks of 20 phenolic compounds were detected in R. rosea leaves, of which 19 compounds were tentatively identified. The leaf extract contained eight phenolic compounds from the group of galloyl-glucoses (three isomers of monogalloyl-glucose, trigalloyl-glucose, two isomers of tetragalloyl-glucose, two isomers of pentagalloyl-glucose), two gallotannins (isomers of hexagalloyl-glucose), five flavonoids (four herbacetin derivatives and one gossypetin), phenolic acids (caffeoyl-malic and ferulic acids), arbutin and esculetin. It has been shown that the biologically active substances of R. rosea provide prospects for the use of leaves as raw materials for the development of herbal preparations with anti-inflammatory, antiviral and antitumor effects.

With the escalating demand for Astragalus polysaccharides products developed from Radix Astragali (RA), the necessity for quality control of polysaccharides in RA has become increasingly urgent. In this study, a specific method for the simultaneous determination of seven monosaccharides in polysaccharides extracted from Radix Astragali (RA) has been developed and validated using ultra-performance liquid chromatography equipped with an ultraviolet detector (UHPLC-UV) for the first time. The 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatizations were separated on a C18 column (Waters ACQUITYTM, Milfor, MA, USA, 1.8 µm, 2.1 × 100 mm) using gradient elution with a binary system of 5 mm ammonium formate (0.1% formic acid)-acetonitrile for 24 min. Additionally, seven monosaccharides showed good linear relationships (R2, 0.9971–0.9995), adequate precision (RSD < 4.21%), and high recoveries (RSD < 4.70%). The established method was used to analyze 109 batches of RA. Results showed that the Astragalus polysaccharides (APSs) mainly consist of mannose (Man), rhamnose (Rha), glucose (Glu), galactose (Gal), arabinose (Ara), xylose (Xyl); and fucose (Fuc); however, their composition was different among RA samples from different growth patterns, species, growth years, and origins, and the growth mode of RA and the age of wild-simulated RA can be accurately distinguished by principal component analysis (PCA). In addition, the immunological activity of APSs were also evaluated jointly by measurement of the NO release with RAW264.7, with the results showing that APSs have a promoting effect on the release of NO and exhibit a significant correlation with Man, Glu, Xyl, and Fuc contents. Accordingly, the new established monosaccharides analytical method and APS-immune activity determination in this study can provide a reference for quality evaluation and the establishment of quality standards for RA. [ABSTRACT FROM AUTHOR]

In the present study, a comprehensive strategy based on substances profiling in vivo and network pharmacology method was applied to screen out the potential immunomodulatory activity substances and mechanism of Wu Shen Wan. As a result, a total of 121 constituents were identified or tentatively characterized, including 26 flavonoids, 16 alkaloids, 10 tanshinones, four diterpenes, seven iridoid glycosides, eight coumarin, 18 phenolic acids, 29 ginsenosides, and three others. Moreover, 57 prototypes were clearly screened out in rat plasma and urine after ingestion of Wu Shen Wan. The Wu Shen Wan exhibited immunomodulatory effects via significantly increasing the phagocytic activity of macrophages and inducing the release of nitric oxide, tumor necrosis factor‐α, and interleukin 6 in RAW264.7 cells. In addition, the network pharmacology results showed that the top 20 Kyoto Encyclopedia of Genes and Genomes clusters were enriched, such as mitogen‐activated protein kinases pathway. This is the first systematic study on the chemical constituents of Wu Shen Wan. The analytical method features a quick and comprehensive dissection of the chemical composition and provides a basis for exploring its immunomodulatory effects. [ABSTRACT FROM AUTHOR]

Tanacetum balsamita L. is a perennial rhizomatous plant of the Asteraceae family. T. balsamita is cultivated as medicinal, aromatic plants and food plant. Extract of costmary exhibits hepatoprotective, antiseptic and anthelmintic properties. The herb of T. balsamita contains essential oils and phenolic compounds. This species included in the "Biocollection of medicinal and aromatic plants of open and protected ground", Botanical garden of the VILAR. The main aim was to study the composition of phenolic compounds in the leaves of T. balsamita with application of ultra-performance liquid chromatography coupled with diode array detection and mass spectrometry. The object of the study was the leaves of T. balsamita. Samples were extracted with 1 ml of 80% aqueous acetone. An ultra-performance liquid chromatographic system with a photodiode detector and a triple quadrupole mass spectrometer was used for analysis of phenolic compounds. The UV and MS data of phenolic compounds were used for their identification or tentative characterization with application of mass spectrometry databases and data published in the literature.Results. The results obtained showed the presence in the leaves of 17 phenolic compounds. Five compounds were identified as caffeoylquinic, feruloylquinic and three dicaffeolquinic acid isomers, and two compounds as ferulic acid hexoside isomers. The leaves of T. balsamita contained also five luteolin derivatives, two chrysoeriol derivatives, and spinacetin/axillarin. Four compounds were not identified.

Purpose: L-citrulline is a non-essential and non-protein amino acid with potent antioxidant properties. In this study, the new aqueous two-phase system (ATPS) was used with and without ultrasound for the first time to isolate and purify L-citrulline from watermelon rind simultaneously. Methods: This system consists of an ionic liquid 1-Butyl-3-methylimidazolium bromide [C4mim] [Br] and dipotassium hydrogen phosphate (K2HPO4) salt. Accordingly, the impact of increasing the salt concentration, the ionic liquid, the solid–liquid ratio, and the ultrasound power and time on the extraction efficiency of L-citrulline and sugar compounds were examined. The HPLC technique Was employed to determine the concentration of L-citrulline in the sample with the highest efficiency rate. Moreover, the DLS tests were applied to determine the particle size. Results: As the concentrations of the ionic liquid, salt, and solid–liquid ratio were increased in the aqueous two-phase extraction system (ATPS) without applying the ultrasonic assistant, the L-citrulline extraction efficiency was increased initially and then decreased. Based on the results, applying the ultrasound to the sample with the highest amino acid efficiency rate, improved the extraction process and enhanced the amino acid extraction efficiency by 90.45%. The best extraction conditions obtained are related to the set, including the 1.5 M ionic liquid, 35% salt, the 1:40 solid–liquid ratio, the power of 100 W, and the ultrasonic time of 15 min. Conclusion: In general, the ionic liquid is a perfect extracting substance, and the ILUAE approach is a simple, efficient, fast, green, and environmentally friendly method for extracting ingredients. [ABSTRACT FROM AUTHOR]

Background: Maple syrup urine disease (MSUD) is a severe life-threatening metabolic disorder. Patients' poor outcomes could be prevented by early diagnosis and regular monitoring, which mainly depend on the analysis of branched amino acids (BCAAs) in plasma. The study aimed to test whether the analysis of BCAAs by ultra-performance liquid chromatography (UPLC) is an alternative to an analysis by ion-exchange chromatography (IEC) for the diagnosis and monitoring of MSUD. Methods: The two methods analyzed fifty plasma samples obtained from treated and untreated patients with MSUD. Data were analyzed using Passing-Bablok and Bland-Altman methods. Results: The slope of the regression lines was equal or close to one for the three BCAAs, indicating no significant proportional differences between the two methods. A slight positive or negative bias was found for leucine and alloisoleucine, respectively. However, for each amino acid, one or two measurement pairs were out of statistical interval of agreement. Despite small analytical differences, the two methods could be considered in clinical agreement since the differences have no impact on the diagnosis and management of patients. Conclusions: UPLC and IEC methods are in clinical agreement for plasma BCAAs analysis. The UPLC method could be used simultaneously or interchangeably with the IEC method for diagnosing and monitoring MSUD patients. However, for reasons of practicability, the alternative method should only be used when the usual method cannot be carried out. [ABSTRACT FROM AUTHOR]

Soil is commonly collected from an outdoor crime scene, and thus it is helpful in linking a suspect and a victim to a crime scene. The chemical profiles of soils can be acquired via chemical instruments such as Ultra-Performance Liquid Chromatography (UPLC). However, the UPLC chromatogram often interferes with an unstable baseline. In this paper, we compared the performance of five baseline correction (BC) algorithms, i.e. asymmetric least squares (AsLS), fill peak, iterative restricted least squares, median window (MW), and modified polynomial fitting, in discriminating 30 chromatograms of brownish soils by five locations of origin, i.e. PP, HK, KU, BL, and KB. The performances of the preprocessed sub-datasets were first visually inspected through the mean chromatograms and then further explored via scores plots of principal component analysis (PCA). Eventually, the predictive performances of the partial least squares-discriminant analysis (PLS-DA) models estimated from 1 000 pairs of training and testing samples (i.e. prepared via iterative random resampling split at 75:25) were studied to identify the best BC method. Mean raw chromatograms of the 10 soil samples were different from each other, with evident fluctuated baselines. AsLS and MW corrected chromatograms demonstrated the most significant improvement compared with the raw counterpart. Meanwhile, the scores plot of PCA revealed that most of the sub-datasets produced three separate clusters. Then, the sub-datasets were modelled via the PLS-DA technique. MW emerged as the excellent BC method based on the mean prediction accuracy estimated using 1 000 pairs of training and testing samples. In conclusion, MW outperformed the other BC methods in correcting the UPLC data of soil. Key points UPLC data of soil interfere with baseline drifts. BC can improve the quality of the pixel-level UPLC data. MW emerges as the most desired algorithm in improving the quality of UPLC data of soil. [ABSTRACT FROM AUTHOR]

The ternary mixture of doxycycline hyclate (DOX), ambroxol hydrochloride (AMB), and ambroxol synthetic precursor and process impurity (IMP) as chemically identified in the British Pharmacopeia as 2-amino-3, 5-dibromobenzaldehyde was examined for the first time using an ultra-performance liquid chromatographic technique. The presented work aimed to develop and validate a green accurate sensitive modern method for determination of this mixture. The obtained UPLC was accomplished by using an UPLC C18 column with mobile phase combination of acetonitrile: water (60:40, v/v) which was flowed at the rate of 0.22 mL per minute. Elution was in an isocratic manner and temperature control set at room temperature. UV was used to identify the effluent, which was injected at a volume of 5.00 µL; 370 nm for DOX and 240 nm for AMB and IMP. In concentration ranges; of (5–150) µg mL−1, linearity of both DOX and AMB was attained and of (0.5–40) µg mL−1, linearity of IMP was attained. Accuracy of this UPLC method was found to be 100.28, 99.66, and 100.36 for DOX, AMB, and IMP, respectively. ICH standards were followed while making the method validation. The existing combined pharmaceutical dosage form was analyzed by the obtained UPLC method for determination of the evaluated drugs successfully and the obtained results expressed as (Recovery % ± SD) were 101.02 ± 1.37 for DOX and 102.22 ± 1.56 for AMB. Also, the statistically significant correlation of our created approach to the reported HPLC method revealed no appreciable differences in terms of accuracy and precision. The developed method's greenness profile was appraised using many greenness metrics, green assessment comparison between the established columnar method and the published ones was made. [ABSTRACT FROM AUTHOR]

With the escalating demand for Astragalus polysaccharides products developed from Radix Astragali (RA), the necessity for quality control of polysaccharides in RA has become increasingly urgent. In this study, a specific method for the simultaneous determination of seven monosaccharides in polysaccharides extracted from Radix Astragali (RA) has been developed and validated using ultra-performance liquid chromatography equipped with an ultraviolet detector (UHPLC-UV) for the first time. The 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatizations were separated on a C18 column (Waters ACQUITYTM, Milfor, MA, USA, 1.8 µm, 2.1 × 100 mm) using gradient elution with a binary system of 5 mm ammonium formate (0.1% formic acid)-acetonitrile for 24 min. Additionally, seven monosaccharides showed good linear relationships (R2, 0.9971–0.9995), adequate precision (RSD < 4.21%), and high recoveries (RSD < 4.70%). The established method was used to analyze 109 batches of RA. Results showed that the Astragalus polysaccharides (APSs) mainly consist of mannose (Man), rhamnose (Rha), glucose (Glu), galactose (Gal), arabinose (Ara), xylose (Xyl); and fucose (Fuc); however, their composition was different among RA samples from different growth patterns, species, growth years, and origins, and the growth mode of RA and the age of wild-simulated RA can be accurately distinguished by principal component analysis (PCA). In addition, the immunological activity of APSs were also evaluated jointly by measurement of the NO release with RAW264.7, with the results showing that APSs have a promoting effect on the release of NO and exhibit a significant correlation with Man, Glu, Xyl, and Fuc contents. Accordingly, the new established monosaccharides analytical method and APS-immune activity determination in this study can provide a reference for quality evaluation and the establishment of quality standards for RA.

A method was established to determine 22 free amino acids in food for special medical purposes by ultra-performance liquid chromatography. After extraction with sulfosalicylic acid and derivation with 2,4-dinitrofluorobenzene (DNFB), the samples were eluted with acetonitrile water-ammonium formate as mobile phase. Amino acids were separated by HSS T3 column and detected with diode-array detector by external standard method. Results showed that, the 22 amino acids were linear in the range of 2~100 µg/mL with correlation coefficients higher than 0.99. The limits of detection (S/N≥3) were 10 mg/100 g, and the limits of quantification (S/N≥10) were 25 mg/100 g. The recoveries were in the range of 90.1% and 109.1% with relative standard deviation (RSD) from 0.8% to 6.9%. This method was simple, efficient, accurate and reproducible. It was suitable for determination of 22 free amino acids in food for special medical purposes, which could provide technical support to ensure the quality and safety.

Hugan tablet is a Chinese medicine preparation. It is composed of Bupleuri Radix, Artemisiae Scopariae Herba, Isatidis Radix, Schisandrae Chinensis Fructus, Suis Fellis Pulvis, and Vigna radiata L. It has the effects of dispersing stagnated liver qi, strengthening the spleen and eliminating food to be used for the treatment of chronic hepatitis and early cirrhosis. However, the chemical composition of Hugan tablet is complex and not fully understood, which hampers the research in pharmacology. In this study, a reliable method for the rapid analysis and identification of the chemical components in Hugan tablet by their characteristic fragments and neutral losses using ultra‐performance liquid chromatography‐quadrupole‐exactive orbitrap mass spectrometry was developed. A total of 144 chemical components were tentatively identified, including 57 organic acids, 19 flavonoids, 23 alkaloids, 18 lignans, 7 saponins, and 20 others. These components may be the active ingredients of Hugan tablet. The established method can systematically and rapidly analyze the chemical components in Hugan tablet, which provides a basis for the pharmacodynamic substance study and is meaningful for the quality control of Hugan tablet. [ABSTRACT FROM AUTHOR]

Background: Chronic renal failure (CRF) results in significant dyslipidemia and profound changes in lipid metabolism. Polyporus umbellatus (PPU) has been shown to prevent kidney injury and subsequent kidney fibrosis. Methods: Lipidomic analysis was performed to explore the intrarenal profile of lipid metabolites and further investigate the effect of PPU and its main bioactive component, ergone, on disorders of lipid metabolism in rats induced by adenine. Univariate and multivariate statistical analyses were performed for choosing intrarenal differential lipid species in CRF rats and the intervening effect of n-hexane extract of PPU and ergone on CRF rats. Results: Compared with control group, decreased creatinine clearance rate indicated declining kidney function in CRF group. Based on the lipidomics, we identified 65 lipid species that showed significant differences between CRF and control groups. The levels of 12 lipid species, especially fatty acyl lipids including docosahexaenoic acid, docosapentaenoic acid (22n-3), 10,11-Dihydro-12R-hydroxy-leukotriene C4, 3-hydroxydodecanoyl carnitine, eicosapentaenoic acid, hypogeic acid and 3-hydroxypentadecanoic acid had a strong linear correlation with creatinine clearance rate, which indicated these lipid species were associated with impaired renal function. In addition, receiver operating characteristics analysis showed that 12 lipid species had high area under the curve values with high sensitivity and specificity for differentiating CRF group from control group. These changes are related to the perturbation of fatty acyl metabolism. Treatment with PPU and ergone improved the impaired kidney function and mitigated renal fibrosis. Both chemometrics and cluster analyses showed that rats treated by PPU and ergone could be separated from CRF rats by using 12 lipid species. Intriguingly, PPU treatment could restore the levels of 12 lipid species, while treatment with ergone could only reverse the changes of six fatty acids in CRF rats. Conclusion: Altered intrarenal fatty acyl metabolites were implicated in pathogenesis of renal fibrosis. PPU and ergone administration alleviated renal fibrosis and partially improved fatty acyl metabolism. These findings suggest that PPU exerted its renoprotective effect by regulating fatty acyl metabolism as a potential biochemical mechanism. Therefore, these findings indicated that fatty acyl metabolism played an important role in renal fibrosis and could be considered as an effective therapeutic avenue against renal fibrosis.

This study dealt with determination of ochratoxin A (OTA) in roasted coffee by ultra-performance liquid chromatography and fluorescence detection (UPLC-FLD). Coffee features characteristic sensory properties, contains a wide range of beneficial substances and is among the most sold commodities worldwide. UPLC-FLD method for determination of OTA in ground roasted coffee was optimized and validated. A calibration curve of good linearity (R² > 0.999) was obtained for the range of 0.3-10 ng·ml-1, and the limit of detection and limit of quantification for OTA were 0.1 µg·kg-1 and 0.3 µg·kg-1, respectively. The recovery of certified reference material for OTA ranged between 81.3-112.8 % and the relative standard deviation under repeatibility conditions was 11.7 %. The method was applied to the analysis of OTA in coffee obtained from retail stores in Czech Republic. Sixteen samples (29 %) of roasted coffee were positive, containing OTA in the range from 0.31 µg·kg-1 to 1.06 µg·kg-1. None of the samples analysed exceeded the legislative limit according to EU Commission Regulation No. 1881/2006. [ABSTRACT FROM AUTHOR]

BACKGROUND: Sulfonamide residues in foods of animal origin are potential risks to consumer health, so it is very important to inspect them. Among the previously reported instrumental methods, the best method can only be used to determine at most 22 sulfonamides. Thus, an instrumental method capable of determining more sulfonamide species is desirable. RESULTS: In this study, sulfadoxine was used as a template to synthesize a type of magnetic molecularly imprinted polymer that could recognize 35 sulfonamides. After characterization, this composite was used to develop a dispersive solid‐phase extraction method for extraction and purification of the 35 sulfonamides in pork, followed by determination using an ultra‐performance liquid chromatography photodiode array method. This composite exhibited high adsorption capacity (11.01–19.21 μg mg−1) and high recovery (>89.01%), and could be reused at least ten times. Due to the enrichment effect during sample preparation (enrichment factor 22–66), the limits of detection for determination of the 35 drugs in pork were in the range of 0.08–0.53 ng g−1. The detection results for some real pork samples were consistent with a liquid chromatographic–tandem mass spectrometric method. After comparison, the present method showed generally better performances than the previously reported sample preparation methods and instrumental methods for detection of sulfonamides. CONCLUSION: The method developed in the present study could be used as a practical tool for routine detection of sulfonamide residues in pork samples. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Overexpression of Glu-1Bx7 via allele 1Bx7OE significantly contributes to high dough strength in some wheat varieties and is useful for improving wheat quality. However, the proportion of wheat varieties containing Bx7OE is quite low. In this study, four cultivars containing 1Bx7OE were selected, and among the selected varieties, Chisholm (1Ax2*, 1Bx7OE + 1By8*, and 1Dx5 + 1Dx10) was crossed with Keumkang, a wheat variety that contains 1Bx7 (1Ax2*, 1Bx7 + 1By8, and 1Dx5 + 1Dx10). SDS-PAGE and UPLC analyses showed that the expression of the high-molecular-weight glutenin subunit (HMW-GS) 1Bx7 was significantly higher in NILs (1Ax2*, 1Bx7OE + 1By8*, and 1Dx5 + 1Dx10) compared with that in Keumkang. Wheat quality was analyzed with near infrared reflectance spectroscopy by measuring the protein content and SDS-sedimentation of NILs. The protein content of NILs (12.94%) was 21.65% higher than that of Chisholm (10.63%) and 4.54% higher than that of Keumkang (12.37%). In addition, the SDS-sedimentation value of NILs (44.29 mL) was 14.97% and 16.44% higher than that of Keumkang (38.52 mL) and Chisholm (38.03 mL), respectively. This study predicts that the quality of domestic wheat can be improved by crossbreeding with 1Bx7OE-containing cultivars. [ABSTRACT FROM AUTHOR]

Chirality plays a vital role in the lives of humans, plants, and animals because various proteins, amino acids, hormones, enzymes are chiral in nature. It is important in different areas such as agriculture, medicine, food, and chemical industries. Chiral compounds may have varying pharmacodynamic and pharmacokinetic parameters, as well as pharmacological and toxic characteristics. Because of their advantages in terms of potency and safety, enantiomerically pure chiral compounds are increasingly being developed by the pharmaceutical industry. This is reinforced by the development of novel methods for enantioselective chiral chemical synthesis as well as the ability of enantioselective analytical methods to identify and quantify small enantiomeric impurities in the presence of a significant excess of another enantiomer. This work aims to offer a short overview on the essential principle of chiral separations on a quantitative scale through this article. In addition, this article includes various types of chiral stationary phases and their applications. Furthermore, gas and liquid chromatography are also discussed as analytical methods used for the separation of chiral compounds. [ABSTRACT FROM AUTHOR]

There is an increased need for the sensitive and accurate measurement of estradiol levels in patients with estradiol-related endocrine disorders. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a reliable and accurate method for measuring steroid hormone levels. Here, we aimed to establish an LC-MS/MS-based method to quantify estradiol levels without sample derivatization and evaluated its analytical performance. Sciex Triple Quad 6500 + LC-MS/MS was used for estradiol analysis. We evaluated its analytical performance, including linearity, precision, the lower limit of detection and quantification (LoD and LoQ, respectively), accuracy, and carryover. The estradiol results determined by LC-MS/MS were compared with those obtained using a chemiluminescent microparticle immunoassay. The LC-MS/MS output was linear for serum estradiol concentrations in the range of 0.2–10311.6 pmol/L. The intra-laboratory precision (coefficient of variation) was 3.0–10.1 %. The LoD and LoQ were 2.8 and 7.5 pmol/L, respectively. The overall accuracy was within 15 % of bias, and the carryover was within the acceptable range (<1.0 %). The results of the estradiol analysis determined by LC-MS/MS were comparable to those obtained by the chemiluminescent microparticle immunoassay (r2 = 0.9843), although there was a negative bias of − 17.82 (95 % confidence interval, −27.21 to − 8.44). A highly sensitive, derivatization-free LC-MS/MS method was successfully developed in this study. This may be beneficial for estradiol measurements in patients with estradiol-related endocrine disorders. [ABSTRACT FROM AUTHOR]

Dendrobium officinale Kimura et Migo is a famous plant with a high medicinal value which has been recorded in the Chinese Pharmacopoeia (2020 Edition). The medicinal properties of D. officinale are based on its chemical composition. However, there are no reports on how different cultivation methods affect its chemical composition. In order to reveal this issue, samples of the D. officinale were collected in this study through tree epiphytic cultivation, stone epiphytic cultivation, and greenhouse cultivation. Polysaccharides were determined by phenol sulfuric acid method and secondary metabolites were detected by the UPLC-MS technique. In addition, with regards to metabolomics, we used multivariate analyses including principal component analysis (PCA) and orthogonal partial least squares analysis (OPLS-DA) to screen for differential metabolites which met the conditions of variable importance projection values >1, fold change >4, and p < 0.05. The differential metabolites were taken further for metabolic pathway enrichment analysis, which was based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and validated by antioxidant activity. Comparing the three groups of samples according to the standards of the ChP (2020 edition), the results showed that the polysaccharide content of the samples from stony epiphytic cultivation and greenhouse cultivation was significantly higher than that of the samples from live tree epiphytic cultivation. Metabolomic analysis revealed that there were 185 differential metabolites among the 3 cultivation methods, with 99 of the differential metabolites being highest in the stone epiphytic cultivation. The results of the metabolic pathway enrichment analysis showed that the different cultivation strategies mainly effected four carbohydrate metabolic pathways, five secondary metabolite synthesis pathways, six amino acid metabolic pathways, one nucleotide metabolism pathway, three cofactor and vitamin metabolism pathways, and one translation pathway in genetic information processing. Furthermore, D. officinale from stone epiphytic cultivation which had the best antioxidant activity was implicated in differential metabolite production. This study revealed the effects of different cultivation methods on the chemical composition of D. officinale and also provided a reference for establishing the quality control standards to aid its development and utilization. [ABSTRACT FROM AUTHOR]

Quaternary benzo[c]phenanthridine alkaloids (QBAs), such as chelerythrine (CHE) and nitidine (NIT), represent the important components to exert the efficacy of Zanthoxylum nitidum (ZN). Ethoxychelerythrine (ECHE), a derivative of CHE, is currently used as a quality marker for the identification of ZN extracts in thin-layer chromatography (TLC) as per Chinese Pharmacopoeia. However, due to the active chemical nature of QBAs, ECHE may be formed as an artifact during sample processing, which leads to the wrong use of quality markers. This study was conducted to investigate whether CHE could be a more scientific and stable quality marker for ZN than ECHE. CHE and ECHE were identified on a TLC using different developing systems. Direct analysis in real-time mass spectrometry (DART-MS) was used to monitor the chemical changes of CHE in different solvents. CHE, ECHE, and corresponding bands of ZN extracts on the TLC plate were determined by TLC‒MS and ultra-performance liquid chromatography (UPLC) coupled with quadrupole time-of-flight mass spectrometry (Q-TOF–MS). The mixture of CHE and ECHE showed a single TLC band corresponding to CHE due to the rapid chemical conversation. Consistently, all the results from DART-MS, TLC‒MS and Q-TOF analysis showed an immediate conversion of ECHE to CHE on the TLC plate. This study strongly suggests that a more stable CHE on TLC plates should be considered to replace ECHE as a quality marker for ZN in the Chinese Pharmacopoeia. [ABSTRACT FROM AUTHOR]

Objective: Granules of herbal extracts are a popular medicinal preparation consumed in traditional Chinese medicine clinical practice. However, their quality and efficacy evaluation are lacking. This study aimed to compare the quality and anti-oxidant activity of Dan Shen (Salvia miltiorrhiza Radix & Rhizoma) granule extracts with their herbal extracts. Methods: Chromatographic method was used to determine the content of 7 marker compounds in the water extracts of the herb compared to that of 12 granule extracts. Agglomerative hierarchical clustering (AHC) and principal component analysis (PCA) distinguished the herbal and granule extracts based on the content of the marker compounds. The antioxidant activities of herbal and granule extracts were evaluated by 2, 2′-azino-bis (3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS), organic chemical compound 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric ion reducing antioxidant power (FRAP) assays. Results: The herbal extracts group showed significantly higher contents of salvianolic acid B, sodium danshensu and cryptotanshinone compared with that of the granule group. This corresponded to significantly higher ABTS, DPPH and FRAP (P

Introduction:Galactosemia (GAL) is a genetic disorder that results in disturbances in galactose metabolism and can lead to life-threatening complications. However, the underlying pathophysiology of long-term complications in GAL remains poorly understood.Methods: In this study, a metabolomics approach using ultra-performance liquid chromatography coupled with high-resolution mass spectrometry was used to investigate metabolomic changes in dried blood spots of 15 patients with GAL and 39 healthy individuals.Results: The study found that 2,819 metabolites underwent significant changes in patients with GAL compared to the control group. 480 human endogenous metabolites were identified, of which 209 and 271 were upregulated and downregulated, respectively. PA (8:0/LTE4) and ganglioside GT1c (d18:0/20:0) metabolites showed the most significant difference between GAL and the healthy group, with an area under the curve of 1 and 0.995, respectively. Additionally, the study identified potential biomarkers for GAL, such as 17-alpha-estradiol-3-glucuronide and 16-alpha-hydroxy DHEA 3-sulfatediphosphate.Conclusion: This metabolomics study deepened the understanding of the pathophysiology of GAL and presented potential biomarkers that might serve as prognostic biomarkers to monitor the progression or support the clinical diagnosis of GAL.

Classical and chemometric calibration strategies based‐Ultra Performance Liquid Chromatography‐Photodiode Array (UPLC‐PDA) measurements were improved to monitor the in‐vitro dissolution profiles of lamivudine (LAM) and zidovudine (ZID) in tablets and to quantify drugs. To generate UPLC‐PDA data, UPLC analysis was done using the mobile phase containing acetonitrile‐acetate buffer pH 5.0 (18 : 82, v/v). In the classical strategy, calibration curves were obtained by using the peak area ratios detected at 274 nm for LAM and 268 nm for ZID, respectively. Principal component regression (PCR) and partial least squares (PLS) strategies were used to solve the same problem using UPLC‐PDA measurements obtained at eight different wavelengths. Both classical and chemometric approaches were successfully applied for drugs assay and in‐vitro dissolution testing. Assay results showed that the proposed chemometric tools gave better results because they helped to minimize the effect of chromatographic issues that commonly occur with the classical UPLC approach, such as the higher standard deviation of the assay results. [ABSTRACT FROM AUTHOR]

The preferred test methods for control of product-related impurities in medicinal products are high-performance liquid chromatography (HPLC) with a fine sorbent, and ultra-performance liquid chromatography (UPLC), which allow for better chromatographic separation of active substances and related impurities, reduction of time costs, and saving of material resources. The aim of the study was to develop HPLC and UPLC test procedures and assess the chromatographic separation capacity and efficiency in order to improve determination of the main vancomycin component and related impurities. Materials and methods: vancomycin hydrochloride lyophilisate for oral solution and solution for injection, and vancomycin hydrochloride reference standard (USP RS) were used as test objects. Agilent 1290 Infinity liquid chromatography system, and Chromolith® Performance RP-18e, Kinetex C18, Nucleodur C18 Isis, Zorbax RRHD Eclipse Plus C18, and LiChrospher® RP-18 columns were used for the testing. Results: HPLC analysis using a Chromolith® column (100×4.6 mm) reduces the testing time by 10 minutes compared to the USP test procedure, and by 15 minutes compared to the British Pharmacopoeia procedure. The proposed test procedure requires less eluent and increases chromatographic separation efficiency. UPLC analysis using a Kinetex C18 column (50×4.6 mm, 2.6 μm) made it possible to reduce the testing time by two thirds compared to the British Pharmacopoeia procedure. The use of isocratic elution greatly simplified the testing. The testing time under the proposed chromatographic conditions was 10 minutes. Conclusions: the selected HPLC and UPLC test conditions made it possible to significantly reduce the time of testing, minimise the use of expensive reagents, and increase efficiency of chromatographic separation in the determination of vancomycin impurities and the main component Vancomycin B.

Gaucher disease is a rare inherited disorder caused by a deficiency of the lysosomal acid beta-glucocerebrosidase enzyme. Metabolomic studies by our group targeted several new potential urinary biomarkers. Apart from lyso-Gb1, these studies highlighted lyso-Gb1 analogs −28, −26, −12 (A/B), +2, +14, +16 (A/B), +30, and +32 Da, and polycyclic lyso-Gb1 analogs 362, 366, 390, and 394 Da. The main objective of the current study was to develop and validate a robust UPLC-MS/MS method to study the urine distribution of these biomarkers in patients. Method: Urine samples were purified using solid-phase extraction. A 12 min UPLC-MS/MS method was developed. Results: Validation assays revealed high precision and accuracy for creatinine and lyso-Gb1. Most lyso-Gb1 analogs had good recovery rates and high intra- and interday precision assays. Biomarker-estimated LOD and LOQ levels ranged from 56–109 pM to 186–354 pM, respectively. Comparison between GD patients and healthy controls showed significant differences in most biomarker levels. Typically, treated GD patients presented lower biomarker levels compared to untreated patients. Conclusions: These data suggest that the metabolites investigated might be interesting GD biomarkers. More studies with a larger cohort of patients will be needed to better understand the clinical significance of these GD biomarkers. [ABSTRACT FROM AUTHOR]

A graphitic carbon nitride was synthesised and nalidixic acid (NA) based molecularly imprinted microspheres (MIMs) were polymerised on its surface to create a composite material. After characterisation and evaluation of its absorption ability, the composite was used to prepare a solid-phase extraction (SPE) cartridge for purification of fluoroquinolones in chicken muscle, analysed by ultra-performance liquid chromatography. The cartridge showed high absorption capacities (378–559 μg) and high recoveries (92.1–99.4%) for eight fluoroquinolones, and could be reused at least 20 times. The limits of detection for the 8 drugs were 0.2–0.8 ng g−1, and recoveries from standard fortified blank chicken muscle samples were 71.9–96.8%. This is the first study reporting the use of molecularly imprinted graphitic carbon nitride composite to determine the residue of veterinary drug in foods of animal origin. [ABSTRACT FROM AUTHOR]

Background & aimsMicrobes may increase susceptibility to inflammatory bowel disease (IBD) by producing bioactive metabolites that affect immune activity and epithelial function. We undertook a family based study to identify microbial and metabolic features of IBD that may represent a predisease risk state when found in healthy first-degree relatives.MethodsTwenty-one families with pediatric IBD were recruited, comprising 26 Crohn's disease patients in clinical remission, 10 ulcerative colitis patients in clinical remission, and 54 healthy siblings/parents. Fecal samples were collected for 16S ribosomal RNA gene sequencing, untargeted liquid chromatography-mass spectrometry metabolomics, and calprotectin measurement. Individuals were grouped into microbial and metabolomics states using Dirichlet multinomial models. Multivariate models were used to identify microbes and metabolites associated with these states.ResultsIndividuals were classified into 2 microbial community types. One was associated with IBD but irrespective of disease status, had lower microbial diversity, and characteristic shifts in microbial composition including increased Enterobacteriaceae, consistent with dysbiosis. This microbial community type was associated similarly with IBD and reduced microbial diversity in an independent pediatric cohort. Individuals also clustered bioinformatically into 2 subsets with shared fecal metabolomics signatures. One metabotype was associated with IBD and was characterized by increased bile acids, taurine, and tryptophan. The IBD-associated microbial and metabolomics states were highly correlated, suggesting that they represented an integrated ecosystem. Healthy relatives with the IBD-associated microbial community type had an increased incidence of elevated fecal calprotectin.ConclusionsHealthy first-degree relatives can have dysbiosis associated with an altered intestinal metabolome that may signify a predisease microbial susceptibility state or subclinical inflammation. Longitudinal prospective studies are required to determine whether these individuals have a clinically significant increased risk for developing IBD.

Pruni Semen, the dried ripe seed of Prunus humilis , P. japonica , or P. pedunculata as recorded in the Chinese Pharmacopoeia, has been widely used in pharmaceutical and food industries. The adulteration of the marketed product with morphologically similar plants of the same genus has led to variable product quality and clinical effectiveness. This study systematically investigated the phylogenetic relationships, morphological traits, and chemical profiles of 37 Pruni Semen samples from planting bases, markets, and fields. DNA barcoding could successfully distinguish the genuine and counterfeit Pruni Semen, and the results indicated that there was almost no authentic Pruni Semen available in the market. The samples were divided into "big seed" (P. pedunculata and P. salicina seeds) and "small seed" (P. humilis , P. japonica, P. tomentosa, and P. avium seeds) categories based on morphology results. The notable discrepancy in the chemical characteristics of "big seed" and "small seed" was that "small seeds" were rich in flavonoids and low in amygdalin, whereas "big seeds" were the opposite. Furthermore, principal component analysis and clustered heatmap analysis verified the distinguishing features of "big seed" and "small seed" based on morphological and chemical characteristics. This study suggested that a combination of DNA barcoding and morphological and chemical characteristics can aid in the identification and quality evaluation of authentic and adulterated Pruni Semen. These findings may help standardize Pruni Semen available in the market and protect the rights and interests of customers. [Display omitted] • Pruni Semen is often adulterated with morphologically similar plants. • Authentic samples were identified and evaluated using DNA barcoding, morphology, and chemical analysis. • No authentic Pruni Semen could be identified in the market. • P. humilis and P. japonica were rich in flavonoids and low in amygdalin, whereas P. pedunculata were the opposite. • Multiforin A was suggested to be used as a quality control indicator for Pruni Semen. [ABSTRACT FROM AUTHOR]

Climate change significantly affects the suitability distributions of natural populations , especially relict species such as Eucommia ulmoides Oliv. (E. ulmoides). However, the driving eco-factors of the distribution of E. ulmoides and the relationship between geo-distribution patterns and quality are still unclear. Here, a new approach was established by combining machine learning models (MaxEnt, Random Forest, and Biomod2 model), ultra-performance liquid chromatography (UPLC) and spatial analysis to address this question. The results reveal that temperature is the most critical variable influencing the distribution of E. ulmoides including Bio1, Bio10, and Bio11, followed by precipitation. Multi-model predictions of suitable habitats for E. ulmoides cover an area ranging from 113.33 × 104 km2 to 285.98 × 104 km2, mainly distributed in the mountainous areas of southwestern China, including the eastern regions of the Dalou, Wuling, and Qinling-Daba Mountains. Under different future scenarios, suitable habitat shifted northward by 184.90–424.88 km, and the contraction area is greater than the expansion area, with the difference ranging from 49.28 × 104 km2 to 98.39 × 104 km2. Eight common components (e.g., chlorogenic acid and pinoresinol doglucoside) and two endemic components (caffeic acid and geniposidic acid) were found in the leaves of the northern and southern medicinal districts (SDM and NMD). The contents of eight key secondary metabolites of E. ulmoides leaves were better distributed in SDM than in NDM. Among these, Xinyang in Henan, Lu'an and Fuyang in Anhui, and the southeastern region of Zhejiang are suitable growing areas for the high-quality cultivation of E. ulmoides , which deserve special attention. The seasonal precipitation (Bio15) was negatively correlated with chlorogenic acid and pinoresinol diglucoside while longitude significantly positively correlated with pinoresinol diglucoside. Overall, this study provides a novel assessment pattern for quality assessment and effective protection of E. ulmoides. [Display omitted] • Assessing the regularity of E. ulmoides geographic and quality distributions using machine learning models and UPLC methods. • Discovering 8 common medicinal components and 2 unique compounds in E.ulmoides leaves from SMD and NMD. • The precipitation seasonality and longitude are vital environmental variables affecting the accumulation of active components in leaves. [ABSTRACT FROM AUTHOR]

Dendrobium officinale Kimura et Migo is a famous plant with a high medicinal value which has been recorded in the Chinese Pharmacopoeia (2020 Edition). The medicinal properties of D. officinale are based on its chemical composition. However, there are no reports on how different cultivation methods affect its chemical composition. In order to reveal this issue, samples of the D. officinale were collected in this study through tree epiphytic cultivation, stone epiphytic cultivation, and greenhouse cultivation. Polysaccharides were determined by phenol sulfuric acid method and secondary metabolites were detected by the UPLC-MS technique. In addition, with regards to metabolomics, we used multivariate analyses including principal component analysis (PCA) and orthogonal partial least squares analysis (OPLS-DA) to screen for differential metabolites which met the conditions of variable importance projection values >1, fold change >4, and p < 0.05. The differential metabolites were taken further for metabolic pathway enrichment analysis, which was based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and validated by antioxidant activity. Comparing the three groups of samples according to the standards of the ChP (2020 edition), the results showed that the polysaccharide content of the samples from stony epiphytic cultivation and greenhouse cultivation was significantly higher than that of the samples from live tree epiphytic cultivation. Metabolomic analysis revealed that there were 185 differential metabolites among the 3 cultivation methods, with 99 of the differential metabolites being highest in the stone epiphytic cultivation. The results of the metabolic pathway enrichment analysis showed that the different cultivation strategies mainly effected four carbohydrate metabolic pathways, five secondary metabolite synthesis pathways, six amino acid metabolic pathways, one nucleotide metabolism pathway, three cofactor and vitamin metabolism pathways, and one translation pathway in genetic information processing. Furthermore, D. officinale from stone epiphytic cultivation which had the best antioxidant activity was implicated in differential metabolite production. This study revealed the effects of different cultivation methods on the chemical composition of D. officinale and also provided a reference for establishing the quality control standards to aid its development and utilization.

Overexpression of Glu-1Bx7 via allele 1Bx7OE significantly contributes to high dough strength in some wheat varieties and is useful for improving wheat quality. However, the proportion of wheat varieties containing Bx7OE is quite low. In this study, four cultivars containing 1Bx7OE were selected, and among the selected varieties, Chisholm (1Ax2*, 1Bx7OE + 1By8*, and 1Dx5 + 1Dx10) was crossed with Keumkang, a wheat variety that contains 1Bx7 (1Ax2*, 1Bx7 + 1By8, and 1Dx5 + 1Dx10). SDS-PAGE and UPLC analyses showed that the expression of the high-molecular-weight glutenin subunit (HMW-GS) 1Bx7 was significantly higher in NILs (1Ax2*, 1Bx7OE + 1By8*, and 1Dx5 + 1Dx10) compared with that in Keumkang. Wheat quality was analyzed with near infrared reflectance spectroscopy by measuring the protein content and SDS-sedimentation of NILs. The protein content of NILs (12.94%) was 21.65% higher than that of Chisholm (10.63%) and 4.54% higher than that of Keumkang (12.37%). In addition, the SDS-sedimentation value of NILs (44.29 mL) was 14.97% and 16.44% higher than that of Keumkang (38.52 mL) and Chisholm (38.03 mL), respectively. This study predicts that the quality of domestic wheat can be improved by crossbreeding with 1Bx7OE-containing cultivars.

Antigenic peptide-loaded cationic liposomes have shown promise as cancer vaccines. Quantification of both peptides and lipids is critical for quality control of such vaccines for clinical translation. In this work we describe a reversed phase ultra-performance liquid chromatography (RP-UPLC) method that separates lipids (DOTAP, DOPC and their degradation products) and two physicochemically different peptides within 12 min. Samples were prepared by dilution in a 1:1 (v/v) mixture of methanol and water. Peptide quantification was done via UV detection and lipids were quantified by an evaporative light scattering detector (ELSD), both coupled to the RP-UPLC system, with high precision (RSD < 3.5%). We showed that the presence of lipids and peptides did not mutually influence their quantification. Limit of detection (LOD) and limit of quantification (LOQ), as determined in the ICH guidelines, were 6 and 20 ng for DOTAP, 12 ng and 40 ng for DOPC, 3.0 ng and 8.0 ng for peptide A and 2.4 ng and 7.2 ng for the more hydrophobic peptide B. Finally, lipid degradation of DOTAP and DOPC was monitored in peptide loaded DOTAP:DOPC liposomes upon storage at 4 °C and 40 °C. [ABSTRACT FROM AUTHOR]

Soil is useful physical evidence for linking a suspect/victim to a particular crime scene. However, the discrimination of soils from neighboring locations can be a very challenging task. In this work, we evaluated four row-wise manipulations (RWMs) on chromatographic data of soils for improving the discriminative ability of the data. Five red, brown, and yellowish-brown soils were collected from five nearby locations in Bangi, Malaysia. Then, the non-volatile organic compounds of soils extracted via acetonitrile were determined using an ultra-performance liquid chromatography (UPLC) system. Eventually, three sets of UPLC data were obtained at 254 nm and independently pre-processed using the four RWMs, respectively. The improvements caused by the RWMs were first derived from visual inspection on the mean chromatograms and scores plot of principal component analysis. Next, the two most desired principal components were determined using a one-way ANOVA-least significant difference test and inspected via composite box plots. Results showed that standard normal variate is the only one that caused the positive impacts though not always. Normalization to sum appeared to reduce the quality of the raw data. Hence, in conclusion, the use of RWMs on the UPLC chromatogram must be carefully evaluated. [ABSTRACT FROM AUTHOR]

Poor chromatographic resolution is one of the main challenges in chromatographic analysis. Partially separated chromatographic peaks frequently occur, due to the nature of analytes and the demand for fast analysis using high flow rates and shorter columns. Modeling of chromatographic three‐way data using suitable chemometric tools enables determining co‐eluted peaks without using additional experimental efforts. In this paper, parallel factor analysis (PARAFAC) was applied to chromatographic data for the quantitative resolution of a quaternary mixture at the co‐elution condition of acetaminophen, aspirin, ascorbic acid, and guaifenesin in a spectrochromatogram. The spectrochromatograms of the calibration set, validation set, and real samples were arranged as a three‐way array. In the next step, the PARAFAC model was implemented to decompose the spectrochromatographic array into trilinear components, corresponding to spectral, chromatographic, and relative concentration profiles of the analytes. The chromatographic and spectral modes were used for the qualitative analysis of components, whereas the analytes in commercial tablets were quantified from their individual profiles in their concentration mode. This study indicated that the application of the PARAFAC model provided a novel strategy for determining overlapping peaks in a chromatogram to perform the analysis of multicomponent mixtures with reduced runtime and without additional efforts. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop a new simple and cost-effective spectrophotometric method to be used as an alternative to chromatography for the analysis of paracetamol and methocarbamol (PAR-MET) combination. In this regard, a comparative study on the quantitative resolution of PAR and MET mixture using continuous wavelet transform (CWT) and ultra-performance liquid chromatography (UPLC) techniques was performed. In interpretation of the spectrophotometric data, three different wavelet families Biorthogonal3.5 (Bior3.5), Mexican Hat Function (Mexh), and Symlets5 (Sym5) were found suitable to get better transform of zero-order spectra of the samples containing PAR and MET. The concentrations of PAR and MET in the working range of 4.0 – 20.0 μg/mL and 5.0 – 45.0 μg/mL, respectively, were proportional to the wavelet amplitudes. This mathematical relationship was used to construct calibration curves for each compound. After validation of the CWT method, analyses of commercial tablets were successfully performed. A new UPLC method was developed and validated in order to compare the results with those obtained by the CWT method and the data were found comparable. The analysis of results obtained from experimental trials showed that the proposed CWTmethod is suitable for the quality control and quantitative assessment of both active substances in tablet formulations. [ABSTRACT FROM AUTHOR]

A method for analyzing fluoroquinolone antibiotics, viz. enrofloxacin and ciprofloxacin, in chicken meat has been developed with the aim of reducing the total analysis time. Certified reference materials (CRMs) for the chicken meat powder, i.e. KRISS CRM 108-03-003 and KRISS CRM 108-03-004, were used for method development. Isotope dilution ultra-performance liquid chromatography–mass spectrometry (ID-UPLC-MS) was employed for sample analysis. The liquid chromatography (LC) conditions were optimized with an "in-sample addition" method using a trace amount of ethylenediaminetetraacetic acid (EDTA), in which EDTA was not added to the mobile phase but to the sample alone. The optimal EDTA amount was determined, and its influence on the separation of the target analyte was investigated. After the feasibility of the "in sample-addition" approach was confirmed, we attempted to improve the previously established sample pretreatment method by varying the conditions for liquid–liquid extraction and solid phase extraction. The developed method was validated using raw chicken meat for recovery evaluation. The "in-sample addition" method afforded highly improved peak shapes without adversely affecting the LC/MS method performance, and the quantification values of enrofloxacin and ciprofloxacin corresponded well with those obtained using the established method. Consequently, the total analysis time for the determination of enrofloxacin and ciprofloxacin in chicken meat CRM could be reduced by simplifying the sample pretreatment procedure. The developed method delivered better reliability compared to the previously established analytical method. The method will be used for stability monitoring of the chicken meat CRM and the development of other meat CRMs for fluoroquinolone analysis and improved food safety applications. [ABSTRACT FROM AUTHOR]

[Display omitted] • Establishment of marker compound ((E)-2-decenedioic acid) that can distinguish natural honey and sugar-fed honey. • Establishment of an analytical method for (E)-2-decenedioic acid in honeys by UPLC. • Validation of the UPLC method by linearity, limit of detection, limit of quantification, precision and accuracy tests. • More amount in sugar-fed honey compared to natural honey contend (E)-2-decenedioic acid. Natural honey is a highly sought-after product owing to its biological uniqueness. We developed a suitable analytical method for the quantitative analysis of ( E )-2-decenedioic acid in Korean natural and sugar-fed honey samples. A total of 258 honey samples were screened using ultra-performance liquid chromatography (UPLC) for the detection and quantification of ( E )-2-decenedioic acid. ( E )-2-decenedioic acid was present in high concentrations (122–127 mg/kg) in sugar-fed honey but only in trace amounts (10.9–14.8 mg/kg) in natural honey. Findings indicate that the UPLC method is suitable and reliable for the quantitative evaluation of ( E )-2-decenedioic acid in honey. We expect that this method will be applicable for detecting honey adulteration. [ABSTRACT FROM AUTHOR]

Background: Therapeutic drug monitoring (TDM) of voriconazole is recommended for personalizing doses. The objective of this study was to compare the enzyme immunoassay developed by ARKTM Diagnostics Inc. for the quantification of voriconazole adapted to the Architect C4000 autoanalyzer (Abbott®) with ultra-performance liquid chromatography using ultraviolet detector (UPLC-UV) method. Materials and Methods: Linearity, precision and accuracy of both methods were validated according to the Food and Drug Administration (FDA) and European Medicines Agency guidelines. The limit of quantification (LOQ) of the UPLC-UV method was determined experimentally. Both methods were applied to the analysis of 62 samples from patients. Correlation was evaluated by Passing-Bablok analysis and the concordance by the Bland–Altman method. Dosage recommendations were generated; the discordances according to the technique were evaluated. Results: All validation parameters determined for UPLC-UV met the criteria set out and LOQ of 0.1 μg/mL was established. However, when the enzyme immunoassay was used to determine concentrations ≤1 μg/ml, CVs were >20%. A linear correlation between both methods was found. However, an overestimation of immunoassay (systematic error of 0.39 μg/mL) was detected. In 11.3% of the samples, the differences in concentrations when they were determined by different techniques would imply a different therapeutic regime. These samples had concentrations close to 1 μg/mL. Conclusion: Although both techniques can be used for TDM of voriconazole, when a value close to the lower limit of the therapeutic range is determined by the ARKTM immunoassay, it would be better to verify the result by a non-automated technique to avoid possible underdosing. [ABSTRACT FROM AUTHOR]

The purpose of this research was to develop a simple, sensitive, and accurate method for simultaneous determination of 35 free amino acids using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Tea samples were extracted with boiling water bath, and then separated by XBridge BEH Amide column by gradient elution. The exact mass and MS/MS spectra of the target compound was detected under the TOF–MS and Information dependent acquisition (IDA)–MS/MS mode. The results demonstrated good linearity (R2 > 0.9980) in the range of 0.5–1,000 ng/mL. The limits of detection (LODs) were 0.13–25.00 mg/kg and the limits of quantitation (LOQs) were 0.25–50.00 mg/kg. The recovery rate ranged from 70.1 to 105.1% with relative standard deviations (RSDs)

Background: Hepatocellular carcinoma (HCC) is the most dangerous complication of chronic liver disease. It is a multifactorial complicated disease. Hepatitis C and hepatitis B viruses (HCV and HBV, respectively) represent the main causes of HCC in Egypt. Early diagnosis is very important to aid in early intervention. Objectives: The goal of this research is to evaluate the metabolic role of different amino acids as non-invasive biomarkers over the course of HCC. Methods: This study included 302 participants with 97 diagnosed, untreated HCC patients, 81 chronic HCV patients, 56 chronic HBV patients, 18 co-infected patients, and a control group of 50 normal age and gender-matched individuals. All participants provided complete medical histories and underwent complete clinical examinations, abdominal ultrasonography and/or computed tomography, routine laboratory investigations, estimation of serum α-fetoprotein, and determination of amino acid levels using ultra-performance liquid chromatography (UPLC MS/MS). Results: This work revealed a decline in branched chain amino acids (BCAA) and increase in aromatic amino acids (AAA) among infected groups (HCC, HBV, HCV, and co-infected patients) compared to control subjects and a marked change in Fisher’s and the BCAAs/tyrosine molar concentration ratios (BTR) between controls and infected groups. Conclusion: Different amino acids could be used as non-invasive markers to discriminate and follow chronic hepatitis patients to predict the course of HCC.

Endogenous metabolites are a class of molecules playing diverse and significant roles in many metabolic pathways for disease. Honokiol (HNK), an active poly-phenolic compound, has shown potent anticancer activities. However, the detailed crucial mechanism regulated by HNK in colorectal cancer remains unclear. In the present study, we investigated the therapeutic effects and the underlying molecular mechanisms of HNK on colorectal cancer in a mouse model (ApcMin/+) by analyzing the urine metabolic profile based on metabolomics, which is a powerful tool for characterizing metabolic disturbances. We found that potential urine biomarkers were involved in the metabolism of compounds such as purines, tyrosines, tryptophans, etc. Moreover, we showed that a total of 27 metabolites were the most contribution biomarkers for intestinal tumors, and we found that the citrate cycle (TCA cycle) was regulated by HNK. In addition, it was suggested that the efficacy of HNK was achieved by affecting the multi-pathway system via influencing relevant metabolic pathways and regulating metabolic function. Our work also showed that high-throughput metabolomics can characterize the regulation of metabolic disorders as a therapeutic strategy to prevent colorectal cancer. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is caused by Mycobacterium tuberculosis and is one of the primary causes of death worldwide. Rapid and accurate diagnosis of TB is one of the most direct means to reduce the incidence of TB. In this study, urinary proteomic profiling of TB patients and non‐TB individual controls (HCs) was performed, and differentially expressed urinary proteins between TB and HCs were compared and exclusively expressed proteins in TB patients were selected to establish a clinically useful disease marker panel. In total, these top 11 targeted proteins with 265 peptides were scheduled for multiple reaction monitoring validation analysis by using urine samples from 52 TB patients and 52 HCs. The result demonstrated that a three‐protein combination out of the five‐protein panel (namely P22352, Q9P121, P15151, Q13291, and Q8NDA2) exhibited sensitivity rate of 82.7% in the diagnosis of TB. Furthermore, the three‐protein combination could differentiate TB from the latent tuberculosis (LTB) effectively, which exhibited specificity rate of 92.3% for the diagnosis of TB from the LTB category. Although more numbers of clinical samples are required for further verification, the results provided preliminary evidence that this "three‐protein combination" out of the five‐protein panel could probably be a novel TB diagnostic biomarker in clinical application. [ABSTRACT FROM AUTHOR]

Hyperlipidemia is a major risk factor for coronary heart disease and has emerged as an important public health problem. Lipidomics is a powerful technology for assessment of global lipid metabolites in a biological system and for biomarker discovery. In the present study, hyperlipidemia was induced by feeding rats a high fat diet. A sensitive ultra-performance liquid chromatography coupled with quadrupole time-of-flight synapt high-definition mass spectrometry method was used for the analysis of plasma lipids. Orthogonal partial least squares-discriminant analysis, correlation analysis and heatmap analysis were performed to investigate the metabolic changes in rats with diet-induced hyperlipidemia. Potential biomarkers were detected using S-plot and were identified by accurate mass data, isotopic pattern and MS(E) fragments information. Significantly increased total cholesterol, triglycerides and low-density lipoprotein cholesterol as well as decreased high-density lipoprotein cholesterol were observed in diet-induced hyperlipidemic rats. Combined with standard serum biochemical results, significant differences in plasma lipid compounds including eleven glycerophospholipids, six fatty acids, two sphingolipids, one eicosanoid, one sterol lipid and one glycerolipid were observed, highlighting the perturbation of lipid metabolism in diet-induced hyperlipidemia. These findings provide further insights into the lipid profile across a wide range of biochemical pathways in diet-induced hyperlipidemia.

Endogenous metabolites are a class of molecules playing diverse and significant roles in many metabolic pathways for disease. Honokiol (HNK), an active poly-phenolic compound, has shown potent anticancer activities. However, the detailed crucial mechanism regulated by HNK in colorectal cancer remains unclear. In the present study, we investigated the therapeutic effects and the underlying molecular mechanisms of HNK on colorectal cancer in a mouse model (ApcMin/+) by analyzing the urine metabolic profile based on metabolomics, which is a powerful tool for characterizing metabolic disturbances. We found that potential urine biomarkers were involved in the metabolism of compounds such as purines, tyrosines, tryptophans, etc. Moreover, we showed that a total of 27 metabolites were the most contribution biomarkers for intestinal tumors, and we found that the citrate cycle (TCA cycle) was regulated by HNK. In addition, it was suggested that the efficacy of HNK was achieved by affecting the multi-pathway system via influencing relevant metabolic pathways and regulating metabolic function. Our work also showed that high-throughput metabolomics can characterize the regulation of metabolic disorders as a therapeutic strategy to prevent colorectal cancer.

As chitin is gaining an increased attention as feedstock for industry, quantification thereof is becoming increasingly important. While gravimetric procedures are long, not specific and highly labour-intensive, acidic hydrolysis of chitin into glucosamine followed by quantification of the latter is more performant. Even though several quantification procedures for the determination of chitin can be found in the literature, they give inconsistent results and their accuracy was not assessed due to the lack of certified analytical standards. Therefore, in the present study, commercially available chitin from practical grade was characterised in detail, allowing the assessment of method accuracy. The procedure for the hydrolysis of chitin into glucosamine and subsequent quantification via UPLC-MS was investigated in detail as well. Using 9-fluorenylmethyl chloroformate (FMOC-Cl) as derivatisation reagent, glucosamine was quantified using reversed-phase chromatography. For the chitin hydrolysis, the highest glucosamine recovery was obtained with 8.0 M HCl for 2 h at 100 °C. The entire procedure for chitin quantification, including the hydrolysis, was characterised by high interday and intraday precision and accuracy. The specificity of the procedure was assessed as well by analysing different mixtures of cellulose and chitin. Chitin recoveries from these analyses ranged from 98.8 to 105.8% while no signal was observed for 100% cellulose, indicating the high specificity of the procedure. It was also concluded that the procedure is much faster and less labour-intensive compared to the gravimetric procedure. [ABSTRACT FROM AUTHOR]

A method is proposed for the identification of surfactants by ultra-performance liquid chromatography (UPLC) with high-resolution mass spectrometry detection after screening water and food samples for the total concentration of cationic and anionic surfactants by microextraction–fluorimetry. The method is based on the use of dispersive liquid–liquid microextraction with chloroform of surfactant ion pairs with organic reagents (eosin and acridine yellow), measuring the fluorescence of the obtained adducts using a smartphone, obtaining RGB colorimetric characteristics, and determining the total surfactant concentration. The main analytical characteristics of the identification of cationic surfactants (alkylpyrdinium, alkyltrimethylammonium, alkyldimethylbenzylammonium (benzalkonium), alkylmethylethylbenzylammonium, didecyldimethylammonium, benzyldimethyl[3-(myristoylamino)propyl]ammonium, N,N-bis(3-aminopropyl)dodecylamine chlorides) and anionic surfactants (alkyl benzene sulfonates (sulfonol), alkyl sulfates, laureth sulfates, alkyl sulfonates, and sodium alkyl carboxylates) by chromatography–mass spectrometry under the selected conditions of chromatographic separation and mass spectrometric detection are found. The features of the chromatographic behavior of the surfactant polymerhomologs under the conditions of UPLC and gradient elution are considered. [ABSTRACT FROM AUTHOR]

Over the past decade, the number of known eicosanoids has expanded immensely and we have now developed an ultra-performance liquid chromatography-electrospray ionization triple quadrupole mass spectrometric (UPLC-QTRAP/MS/MS) method to monitor and quantify numerous eicosanoids. The UPLC-QTRAP/MS/MS approach utilizes scheduled multiple reaction monitoring (MRM) to optimize sensitivity, number of metabolites that can be analyzed and the time requirement of the analysis. A total of 184 eicosanoids including 26 deuterated internal standards can be separated and monitored in a single 5min UPLC run. To demonstrate a practical application, human plasma samples were analyzed following solid-phase extraction (SPE) and the recovery rate and matrix effects were determined for the 26 deuterated internal standards added to the plasma. The method was validated and shown to be sensitive with the limit of quantitation at pg levels for most compounds, accurate with recovery rates of 70-120%, and precise with a CV