Liquid Chromatography-photodiode Array Detection Research Articles

Development of amphiphilic hypercrosslinked porous polymers for magnetic extraction of multiple environmental pollutants in water

Under the principle of similar compatibility, researchers have developed various polarity extractants corresponding to a class of chemicals. Separating different polarities chemicals with one extractant effectively has become a novel research trend in separation science. Given the complexity of environmental sample matrices and the significant differences in polarity and solubility of various compounds, the introduction of hydrophilic groups to hydrophobic material skeletons can lead to sorbents with hydrophilic-lipophilic balance (HLB) property and thus improve their extraction performance for substances with different polarities. In this work, a hypercrosslinked polymer (HCPPz-TPB), designated as HLB, was synthesized by incorporating polar pyrazine and nonpolar triphenylbenzene molecules within each other. Subsequently, a core-shell magnetic composite material was obtained by encapsulating magnetic Fe3O4 nanoparticles in HCPPz-TPB. The material was applied as an adsorbent for magnetic solid phase extraction (MSPE) and combined with a high-performance liquid chromatography-photodiode array detector (HPLC-PDA) to enrich, separate, and detect seven polar contaminants in environmental water samples. The proposed approach, Fe3O4@SiO2@HCPPz-TPB−MSPE-HPLC-PDA, is characterized by its outstanding high sensitivity, low detection limits, wide linear range, and good reproducibility. The method demonstrated satisfactory linearity in the range of 0.05–2 μg mL-1 with R2 values between 0.9969 and 0.9997; the limits of detection (LOD) were observed to be within the range of 0.0019–0.016 μg L-1, and limits of quantification (LOQ) was observed to be within the range of 0.0064–0.054 μg L-1 range with good precision. The recoveries of the different contaminants in the environmental samples ranged from 83.61 to 116.46% (RSD≤10.56, n = 5). The new hydrophilic-lipophilic balance extractant is highly efficient, sensitive, and precise for extracting different polar pollutants. The findings demonstrate that the Fe3O4@SiO2@HCPPz-TPB display a remarkable affinity for multiple targets, driven by complex interactions including multi-stackings and hydrogen bonding as a sorbent. The synthesized Fe3O4@SiO2@HCPPz-TPB may be employed in diverse applications, including extraction, removal, and determination of diverse trace multi-target analytes in complex media.

Circulation of COVID-19-Related Medicines on Japanese Websites during the COVID-19 Pandemic and Their Quality and Authenticity.

Substandard and falsified medical products for treating COVID-19 have spread worldwide. These medicines have entered Japan through personal importation of products purchased via the Internet. In this study, we investigated the circulation of 19 COVID-19-related medicines on the Internet in Japan and evaluated the pharmaceutical quality and authenticity of 2 medicines (dexamethasone tablets and ivermectin tablets) obtained online. We purchased 23 samples of 0.5-mg dexamethasone tablets and 13 samples of 3-mg ivermectin tablets from the Internet in December 2020 and July 2022. We investigated the quality and authenticity of the obtained samples through visual observation and tested their authenticity. We conducted pharmacopoeia compliance testing (quantitative assay, content uniformity tests, and dissolution tests) using the high-performance liquid chromatography-photodiode array detector method. No prescription was ever required at the time of purchase. Visual observation revealed that most samples lacked a package insert and some samples had packaging deficiencies. In terms of authenticity, eight ivermectin samples were genuine; the authenticity of the other samples remained uncertain. Four dexamethasone samples and three ivermectin samples failed quality testing based on pharmacopeia validation standards. Our findings illustrate that dexamethasone and ivermectin tablets of poor quality are available online. It is important to increase consumer awareness and provide information about these medicines to prevent the purchase of substandard medicines via the Internet.

Application of a kosmotrope (Na2CO3) and chaotrope (NaCl) in chemometric optimization of aqueous two-phase extraction of bioactive compounds in Hypoxis iridifolia.

Enterolactone, coumaric acid and vitexin are polyphenolic compounds present in a variety of fruits, vegetables, cereals and plants. These bioactive compounds are in high demand due to their antioxidant property in various tissues and organs. The purpose of this study was to develop a simultaneous extraction method, an aqueous two-phase extraction (ATPE) method, that would enable the extraction of these compounds from Hypoxis iridifolia. This environmentally friendly extraction method only applied water and ethanol as extraction solvents for these analytes from the plant matrix. After phase separation, the analytes were salted-out from the aqueous phase into the organic phase with the aid of a chaotrope (NaCl) or kosmotrope (Na2CO3). Thereafter, the analytes were withdrawn by a micro-pipette for analysis on the high-performance liquid chromatography-photodiode array detector. Optimization was conducted using a central composite design, where three parameters were examined which involved percentage ethanol, centrifugation time and salt type. Generally, the optimized conditions for extraction were an ethanol percentage of 100% and a centrifugation time of 10min, which yielded concentrations of 2942, 23,823 and 8881mg kg-1 for enterolactone, vitexin and coumaric acid, respectively, in the presence of a kosmotrope. The optimized conditions of extraction in the presence of chaotrope were an ethanol percentage of 66% and a centrifugation time of 10min with concentrations of 6727, 20,833 and 8618mg kg-1 for enterolactone, vitexin and coumaric acid, respectively. The ATPE method involving Na2CO3 was a better extractant of all the compounds studied relative to that of NaCl. The superior extraction capability of Na2CO3 in ATPE could serve as a prototype for the development of efficient extraction methods to meet the high demand for medicinal compounds derived from natural products.

On-site sensing for aflatoxicosis poisoning via ultraviolet excitable aptasensor based on fluorinated ethylene propylene strip: a promising forensic tool

The environmental contamination by extremophile Aspergillus species, i.e., Aflatoxin B1, is hardly controllable in Southeast Asia and Sub-Saharan Africa, which lack handling resources and controlled storage facilities. Acute aflatoxicosis poisoning from aflatoxin-prone dietary staples could cause acute hepatic necrosis, acute liver failure, and death. Here, as the cheaper, more straightforward, and facile on-site diagnostic kit is needed, we report an ultraviolet-excitable optical aptasensor based on a fluorinated ethylene propylene film strip. Molecular dynamics on the aptamer.AFB1 complex revealed that the AFB1 to the aptamer increases the overall structural stability, suggesting that the aptamer design is suitable for the intended application. Under various influencing factors, the proposed label-free strategy offers a fast 20-min on-site fabrication simplicity and 19-day shelf-life. The one-pot incubation provides an alternative to catalytic detection and exhibited 4 times reusability. The recovery of crude brown sugar, processed peanuts, and long-grain rice were 102.74 ± 0.41 (n = 3), 86.90 ± 3.38 (n = 3), and 98.50 ± 0.42 (n = 3), comparable to High-Performance Liquid Chromatography-Photodiode Array Detector results. This study is novel owing to the peculiar UV-active spectrum fingerprint and the convenient use of hydrophobic film strips that could promote breakthrough innovations and new frontiers for on-site/forensic detection of environmental pollutants.

Pharmacokinetics of oxytetracycline in hybrid catfish (Clarias macrocephalus x C. gariepinus) after intravascular and oral administrations.

Over the past decade, catfish farming has increased in Southeast Asia. However, there has been no existing for pharmacokinetic data in the hybrid catfish (Clarias macrocephalus x C. gariepinus). This study was designed to evaluate the pharmacokinetic characteristics of oxytetracycline (OTC) in the hybrid catfish, following single intravascular (IV) or oral (PO) administration at a single dosage of 50 mg/kg body weight (BW). In total, 140 catfish (each about 100-120 g BW) were divided into two groups (n = 70). Blood samples (0.6-0.8 mL) were collected from ventral caudal vein at pre-assigned times up to 144 h (sparse samples design). OTC plasma concentrations were analyzed using high-performance liquid chromatography-photodiode array detector. The pharmacokinetic parameter of OTC was evaluated using a non-compartment model. OTC plasma concentrations were detectable for up to 144 and 120 h after IV and PO, respectively. The elimination half-life value of OTC was long with slow clearance after IV administration in hybrid catfish. The average maximum concentration value of OTC was 2.72 µg/mL with a time at the maximum concentration of 8 h. The absolute PO bioavailability was low (2.47%). These results showed that PO administration of OTC at a dosage of 50 mg/kg BW was unlikely to be effective for clinical use in catfish. The pharmacodynamic properties and clinical efficacy of OTC after multiple medicated feed are warranted.

Simultaneous Analysis of Thirteen Compounds in Yeokwisan Using High-Performance Liquid Chromatography-Photodiode Array Detection and Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry and Their Antioxidant Effects.

Yeokwisan (YWS) is an herbal medicine prescription consisting of six oriental herbal medicines, developed to treat reflux esophagitis. We focused on developing an analytical method capable of simultaneously quantifying 13 compounds in YWS samples using high-performance liquid chromatography-photodiode array detection (HPLC-PDA) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and exploring their antioxidant effects. All compounds examined in both analytical systems were chromatographically separated on a SunFireTM C18 (4.6 × 250 mm, 5 μm) column and an Acquity UPLC BEH C18 (2.1 × 100 mm, 1.7 μm) column using gradient elution of a water-acetonitrile mobile phase. Antioxidant effects were evaluated based on radical scavenging activity (DPPH and ABTS tests) and ferrous ion chelating activity. In two analytical methods, the coefficient of determination of the regression equation was ≥0.9965, the recovery range was 81.11-108.21% (relative standard deviation (RSD) ≤ 9.33%), and the precision was RSD ≤ 11.10%. Application of the optimized analysis conditions gave quantitative analysis results for YWS samples of 0.02-100.36 mg/g. Evaluation of the antioxidant effects revealed that baicalein and baicalin exhibit significant antioxidant activity, suggesting that they play an important role in the antioxidant effects of YWS.

First analytical confirmation of drug-induced crystal nephropathy in felines caused by GS-441524, the active metabolite of Remdesivir

GS-441524 is an adenosine nucleoside antiviral demonstrating significant efficacy in the treatment of feline infectious peritonitis (FIP), an otherwise fatal illness, resulting from infection with feline coronavirus. However, following the emergence of COVID-19, veterinary development was halted, and Gilead pursued clinical development of a GS-441524 pro-drug, resulting in the approval of Remdesivir under an FDA emergency use authorization. Despite lack of regulatory approval, GS-441524 is available without a prescription through various unlicensed online distributors and is commonly purchased by pet owners for the treatment of FIP. Herein, we report data obtained from the analytical characterization of two feline renal calculi, demonstrating the propensity for GS-441524 to cause renal toxicity through drug-induced crystal nephropathy in vivo. As definitive diagnosis of drug-induced crystal nephropathy requires confirmation of the lithogenic material to accurately attribute a mechanism of toxicity, renal stone composition and crystalline matrix were characterized using ultra-performance liquid chromatography photodiode array detection (UPLC-PDA), ultra-performance liquid chromatography mass spectrometry (LCMS), nuclear magnetic resonance (NMR) spectroscopy, X-ray powder diffraction (XRD), and Fourier-transform infrared spectroscopy (FTIR). This work serves to provide the first analytical confirmation of GS-441524-induced crystal nephropathy in an effort to support toxicologic identification of adverse renal effects caused by administration of GS-441524 or any pro-drug thereof.

Simultaneous Quantification of Nine Target Compounds in Traditional Korean Medicine, Bopyeo-Tang, Using High-Performance Liquid Chromatography-Photodiode Array Detector and Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Bopyeo-tang (BPT) is composed of six medicinal herbs (Morus alba L., Rehmannia glutinosa (Gaertn.) DC., Panax ginseng C.A.Mey., Aster tataricus L.f., Astragalus propinquus Schischkin, and Schisandra chinensis (Turcz.) Baill.) and has been used for the treatment of lung diseases. This study focused on establishing an analytical method that can simultaneously quantify nine target compounds (i.e., hydroxymethylfurfural, mulberroside A, chlorogenic acid, calycosin-7-O-glucoside, 3,5-dicaffeoylquinic acid, quercetin, kaempferol, schizandrin, and gomisin A) from a BPT sample using high-performance liquid chromatography with a photodiode array detector (HPLC-PDA) and ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). The separation of compounds in both analyses was performed on a C18 reversed-phase column using the gradient elution of water-acetonitrile as the mobile phase. In particular, the multiple reaction monitoring mode was applied for quick and accurate detection in UPLC-MS/MS analysis. As a result of analyzing the two methods, HPLC-PDA and UPLC-MS/MS, the coefficient of determination of the regression equation for each compound was ≥0.9952, and recovery was 85.99-106.40% (relative standard deviation (RSD) < 9.58%). Precision testing of the nine compounds was verified (RSD < 10.0%). The application of these analytical assays under optimized conditions for quantitative analysis of the BPT sample gave 0.01-4.70 mg/g. Therefore, these two assays could be used successfully to gather basic data for clinical research and the quality control of BPT.

A UPLC method development and validation study of Upadacitinib and its impurities in extended – release oral tablet dosage forms

The primary objective was to develop a concomitant isocratic ultra-performance liquid chromatographic photo-diode array detection method to estimate Upadacitinib and its process-related impurities: impurity-1 and impurity-2. Further validation was conducted and studied for possible degradants under stress environments. All the chemicals and reagents used were of HPLC (acetonitrile, methanol) and analytical grade (trifluoro acetic acid). The ultra-performance liquid chromatography (Agilent 1290 Infinity II LC system) consists of a quaternary pump, a BEH C18 (50×2.1mm, 1.7μ) column, and photo-diode array detector. The method was developed with acetonitrile: methanol: 0.1% v/v trifluoro acetic acid (50:20:30 v/v/v) mobile phase at 0.2mL/min flow rate within a run time of 5.5min The detection was carried at 231.2nm. The respective retention times achieved were 2.289min (Upadacitinib), 0.972min (Upadacitinib impurity-1), and 3.508min (Upadacitinib impurity-2). The optimized method was validated further, and the linearity range was best fit at 15.0-180.0μg/mL for Upadacitinib and 1.0-12.0μg/mL for both Upadacitinib impurity-1 and 2 respectively. The detection and quantification limits were 4.50μg/mL, 15.00μg/mL (Upadacitinib) and 0.30μg/mL, 1.0μg/mL (Upadacitinib impurity-1 and 2). A fast, isocratic, specific, and reproducible ultra-performance liquid chromatographic method was developed and validated for various parameters according to the ICH Q2 (R1) guidelines studies. Stress studies were conducted exposing the sample dilution to various treatments (acid, alkali, peroxide, HPLC water, heat, and UV light). The degradants were well-separated apart from the peaks of the active substance. The stability indicating nature was observed during the degradation. The optimized method can be applied for the separation and estimation of Upadacitinib and its process-related impurities in pharma sector in tablet dosage forms.

Titania-based fabric phase sorptive extraction approach for the determination of antiepileptic drugs, levetiracetam and lamotrigine in urine samples using high-performance liquid chromatography-photo diode array detection

A new fabric phase sorptive extraction (FPSE) based separation and enrichment method was developed for sensitive determination of two antiepileptic drug molecules, Levetiracetam (LEV) and Lamotrigine (LTG). The analysis of these drug molecules was performed with high-performance liquid chromatography equipped with photodiode array detector (HPLC-PDA) after FPSE. HPLC analysis was carried out by using phenyl hexyl column, under isocratic conditions with the mobile phase composed of pH 3.0 buffer−acetonitrile (77:23 v: v). All parameters affecting the separation and enrichment process were studied and optimized step by step. The linear working range of the developed method was calculated in the range of 10.0–1000.0 ng mL−1 for both the drug molecules (LEV and LTG). The limits of detection of the method (LODs) were calculated as 2.72 and 3.64 ng mL−1, respectively. The relative standard deviation (%RSD) values of the developed method as an indicator of precision were varied between 4.0 and 7.3. The accuracy of the optimized FPSE method was determined by the recovery tests utilizing spiked samples and results were assessed in the range from 94.6 to 106.3%. This is the first application of sol-gel Titania polycaprolactone-polydimethylsiloxane-polycaprolactone (Ti-PCAP-PDMS-PCAP) based FPSE membrane in the determination of antiepileptic drug molecules.

Dispersive micro solid phase extraction of glibenclamide from plasma, urine, and wastewater using a magnetic molecularly imprinted polymer followed by its determination by a high-performance liquid chromatography-photodiode array detector.

The present study describes the development of a simple and selective analytical method for dispersive micro solid phase extraction and determination of glibenclamide (GLB) using magnetic molecularly imprinted polymer (MMIP) as a sorbent. MMIP was fabricated by the non-covalent method on the surface of silicated Fe3O4 and had a high affinity for glibenclamide; dual monomers, itaconic acid and allylamine, were used for this. Polymerization was achieved by the precipitation method in the presence of glibenclamide as the template and ethylene glycol dimethacrylate as the cross-linker. The morphology and structural properties of the MMIP were characterized by different analytical methods. To achieve maximum extraction efficiency, influencing parameters were optimized. The linearity range was 1-2000 and 12-2000 μg L-1 by high-performance liquid chromatography-photodiode array detector (HPLC-PDA) and UV-vis spectroscopy, respectively. The detection and quantification limits with UV-vis and HPLC-PDA analyses were 4 and 12 μg L-1 and 0.3 and 1 μg L-1, respectively. Under optimized conditions, recovery of glibenclamide spiked in plasma, human urine, and wastewater was between 89.4 and 102.9% at the concentration levels of 25, 250, and 500 μg L-1; relative standard deviations were below 3.7% by HPLC-PDA. The developed method has a favorable pre-concentration factor of 140.0. Equilibrium data and sorption isotherms fitted well with the Langmuir model. A maximum sorption capacity of 24.260 mg g-1 was acquired based on the Langmuir model. The synthesized sorbent with high selectivity was used to separate GLB from complex biological systems and wastewater before measurement with UV-vis or HPLC-PDA.

Analysis of polyphenols and anti-oxidant activity of Punarnava (Boerhavia diffusa L.) from Himachal Pradesh-A North-Western State of India

Boerhavia diffusa L. (family Nyctaginaceae) is also known as Punarnava in Indian medicine and has been used as rasayana herb in Ayurvedic medicine since ancient times. Plant has anti-aging, rejuvenating, cell-strengthening, mind-enhancing and preventative properties. The present study aims to investigate the different plant parts of the species B. diffusa for phytochemicals and antioxidants characterization for the first time from Bilaspur (31.33 oN 76.75 oE), Himachal Pradesh of North-Western India. The methanol: water extracts (leaf, stem and root) of B. diffusa were tested for in vitro antioxidant activity by various assays such as DPPH (1, 1-diphenyl-2-picrylhydrazyl), ABTS [2, 2’-azino-bis (3-ethyl benzothiazoline-6-sulphonic acid)], FRAP (Ferric reducing antioxidant power assay) and RP-HPLC-PDA (Reverse Phase-High Performance Liquid Chromatography-Photo Diode Array Detector), which showed that B. diffusa root had the highest antioxidant activity i.e. DPPH (93.11±0.152g %), ABTS (89.70±0.207g %) and FRAP (88.67±0.251g %). This suggests that plant-derived polyphenols are good sources of natural antioxidants. Further, in the present study, three polyphenols, such as rutin, syringic acid, and epicatechin, were also determined through RP-HPLC-PDA method and found to be maximum in root extract. The results indicated that the methanolic extract of B. diffusa could be a potential herbal medicine against free radicals.

Screening of lupeol, mangiferin and β-carotene contents in pulp of mango (Mangifera indica L.) varieties at edible ripe stage

Mango fruits are an amazing source of numerous bioactive phytonutrients like lupeol (a novel anti-inflammatory and anticancer dietary triterpene), mangiferin (an antidiabetic, anti-HIV, anticancer, immunomodulatory and anti-inflammatory agent) and β-carotene (a significant carotenoid that functions as a precursor to vitamin A, the conscientious reason for our vision). In the present study, ripe pulp of 23 mango varieties was examined for their contents of lupeol, mangiferin and β-carotene using high performance liquid chromatography-photodiode array detector (HPLC-PAD). Mulgoa had the highest lupeol concentration, measuring 42.52 µg/g and Langra stands second with 36.33 µg/g followed by Pairi (33.56 µg/g), Sensation (28.69 µg/g) and Dashehari with 28.22 µg/g. Mangiferin content was highest in variety Arunika (49.58 µg/g) followed by Ambika (34.80 µg/g), Dashehari (33.31 µg/g), Sensation (29.66 µg/g) and Neelum (27.93 µg/g). Sensation's mature pulp has the highest quantity of β-carotene (109.58 µg/g), followed by Kesar (96.87 µg/g), Dashehari (82.13 µg/g), Mulgoa (79.99 µg/g), Arunika (74.26 µg/g), and Amrapali (70.12 µg/g). The pulp of Dashehari, Sensation, Mulgoa, Arunika, Kesar and Amrapali possessed good to moderate amount of these nutraceuticals and are beneficial for consumption at ripe stage. This study has showed the importance of nutraceutical components present in mango; meanwhile it also encourages mango growers to grow these varieties for better profitability.

Influence of industrial blanching, cutting, and freezing treatments on in vitro gastrointestinal digestion stability of orange (Citrus sinensis L.) and lemon (Citrus limon L.) peel polyphenols.

Citrus peels, which are often discarded as waste in the food-processing industry, are rich sources of polyphenols. The aim of the current study was to investigate the influence of an industrial freezing process along with blanching and cutting pretreatments on the in vitro gastrointestinal digestion stability of orange (Citrus sinensis L.) and lemon (Citrus limon L.) peel polyphenols. The major polyphenols were identified and quantified with ultra-performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-ESI-MS/MS) and high performance liquid chromatography-photodiode array detector (HPLC-PDA), respectively. The results revealed that hesperidin and chlorogenic acid were the predominant flavonoid and phenolic acids in orange peels whereas, for lemon peels, eriocitrin and caffeic acid were the dominant flavonoid and phenolic acids, respectively. Blanching pretreatment enhanced the levels of major flavonoids in orange and lemon peels (by 39-82% and 54-146%, respectively) (P < 0.05) after in vitro gastrointestinal digestion. On the other hand, the application of cutting treatment after blanching significantly reduced the levels of flavonoids (23-62%) (P < 0.05); however, there was no statistically significant difference between the phenolic acid levels of cut and uncut citrus peels. Overall, the bioaccessibility of individual flavonoids and phenolic acids from frozen orange peels was generally slightly lower than that of untreated peels (9-34% and 9-49%, respectively). Nevertheless, frozen lemon peels contained higher bioaccessible flavonoids and phenolic acids in comparison with untreated peels (40-172% and 32-98%, respectively). These results suggest that industrial freezing steps could largely preserve the bioaccessibility of polyphenols in orange and lemon peels. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

UPLC-PDA Approach for Simultaneous Quantification of Elvitegravir, Tenofovir, Emtricitabine and Cobicistat in Tablets: An MS-ESI Study for Degradation Products.

A simultaneous ultra-performance liquid chromatography photodiode array detection method was developed and validated for Elvitegravir, Tenofovir and Emtricitabine and Cobicistat drugs in tablet dosage forms. Effective liquid chromatographic quantification was achieved using a Luna C18 (100 × 2.6mm, 1.6μ) column with an acetonitrile: 0.1% v/v trifluoro acetic acid (80:20% v/v) mobile phase and a photo diode array detector at 260nm for a shorter run time of 3min. The respective retention times for Elvitegravir, Tenofovir, Emtricitabine and Cobicistat were 0.311, 0.792, 1.379 and 2.045min. The range of linearity was 37.50-225.00, 2.50-15.00, 50.00-300.00and 37.50-225.00μg/mL for Elvitegravir, Tenofovir, Emtricitabine and Cobicistat, respectively. A mass spectrometer was employed for quantitative analysis using electron spray ionization multiple reaction monitoring in positive mode. The developed method was validated for different parameters to meet the acceptance criteria as per ICH Q2 (R1) guidelines. Using UPLC-ESI-MS, 12 degradants were identified and their mechanisms were studied.

Assessment of Total Phenolic Compounds, Antioxidant Capacity, β-Carotene Bioaccessibility, HMF Formation, and Color Degradation Kinetics in Pumpkin Pestils

Pestil, often known as fruit leather, is one of the most significant traditional foods manufactured and consumed throughout Türkiye. Due to its practical consumption, the availability of numerous nutrients, and the ability to meet energy requirements, pestil is recognized as a snack food. The aim of this study was to evaluate the bioaccessibility of total phenolic compounds (TPC), antioxidant capacity (AOC), and β-carotene in pumpkin pestils dried by hot air drying (HAD), vacuum drying (VCD), and microwave drying (MD) methods using an in vitro digestion model. Additionally, 5-hydroxymethylfurfural (HMF) formation and color degradation of pestils were evaluated. Changes in TPC and AOC were determined using spectrophotometric methods, whereas the detections of β-carotene and HMF were carried out with high performance liquid chromatography–photodiode array detector (HPLC–PDA). Significantly higher TPC (10.99–105.70%) and AOC (15.30–118.58%, 21.88–401.04% and 89.28–482.14%, in CUPRAC, FRAP, and DPPH assays, respectively) values were observed after drying (p&amp;lt;0.05). Moreover, it was observed that there were statistically significant increases in TPC and AOC values after digestion for all pumpkin pestils compared to undigested samples (p&amp;lt;0.05). Drying process resulted in lower β-carotene content (between 32.15–61.11%) in pumpkin pestils; however, it increased the percentage of bioaccessible β-carotene (max 62.16%) in the pestil samples. Compared to HD and VCD techniques, pumpkin pestils dried with MD exhibited significantly higher TPC, AOC and β-carotene content (p&amp;lt;0.05). All of the pumpkin pestils except those dried by MD at 180 W contain HMF below the Turkish Standards Institute legal limit of 50 mg/kg. L* value of pestils were described adequately to the zero- and first-order kinetic models while a* and b* values were only fitted to zero-order model. In conclusion, the findings obtained in this study pointed out that drying processes (especially by MD method) increased the bioaccessibility of TPC, AOC, and β-carotene.

Comparative analysis of fatty acids, volatile and non-volatile components in red huajiao (Zanthoxylum bungeanum maxim.) and green huajiao (Zanthoxylum armatum DC.) using GC-MS, UPLC-LTQ-Orbitrap-MS/MS and HPLC-DAD

In China, huajiao, a traditional industrial crop, is categorized as red huajiao (RHJ; Zanthoxylum bungeanum maxim.) and green huajiao (GHJ; Zanthoxylum armatum DC.) based on the peel color. There were significant differences in sensory and biological activities between RHJ and GHJ in practical application. However, there is no sufficient and comprehensive metabolome-based explanation for these differences so far. The gas chromatography-mass spectrometry (GC-MS), ultra-performance liquid chromatography-linear trap quadrupole-orbitrap mass spectrometry (UPLC-LTQ-Orbitrap-MS/MS), and high-performance liquid chromatography-photodiode array detection (HPLC-DAD) were employed in this study to analyze the differences in fatty acids (FAs), volatile components (VCs) and non-volatile components (non-VCs) between 6 RHJ and 9 GHJ. The findings demonstrated that 12 FAs, 68 VCs, and 101 non-VCs were detected in huajiao, of which 5 FAs, 28 VCs, and 46 non-VCs were identified as differential metabolites (DMs) between RHJ and GHJ. And the DMs were enriched in multiple metabolic pathways by KEGG analysis. Moreover, kaempferol, kaempferide, berberrubine, scoulerine, saponaretin, naringenin, and 3-P-coumaroylquinic acid were identified as key bioactive DMs based on the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and analysis platform. In this study, the possible reasons for the differences in sensory and biological activities between RHJ and GHJ were comprehensively revealed at the metabolic level using non-targeted and targeted metabolomics, and the results provide a better theoretical understanding of the different application of them.

Quantitative-HPLC-DAD polyphenols analysis, anxiolytic and cognition enhancing potentials of Sorbaria tomentosa Lindl. Rehder

Ethnopharmacological relevanceMedicinal plants of the family Rosaceae have a long history of traditional uses in the management of neurological disorders. Sorbaria tomentosa Lindl. Rehder is composed of antioxidant and neuroprotective polyphenolics. Aims of the studyThe current study was designed to explore phenolics profile via high performance liquid chromatography-photodiode array detector (HPLC-DAD) and validated the neuroprotective and anxiolytic potentials of S. tomentosa by applying in vitro and in vivo approaches. Materials and methodsThe plant crude methanolic extract (St.Crm) and fractions were subjected to HPLC-DAD analysis for qualitative and quantitative assessment of phytochemicals. Samples were screened for in vitro free radicals scavenging assays by using 2,2-diphenylpicrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) along with acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes inhibition assays. For cognitive and anxiolytic studies, mice were subjected to open field, elevated plus maze (EPM), light-dark model, Y-maze, shallow water maze (SWM), and novel object recognition (NOR) tests. ResultsHPLC-DAD analysis revealed the presence of high concentrations of phenolic compounds. For instance, in St.Cr, 21 phenolics were quantified, among which apigenin-7-glucoside (291.6 mg/g), quercetin (122.1 mg/g), quercetin-3-feruloylsophoroside-7-glucoside (52.6 mg/g), quercetin-7-glucoside (51.8 mg/g), ellagic acid (42.7 mg/g), luteolin (45.0 mg/g), kaempferol (40.5 mg/g), 5-feruloylquinic acid (43.7 mg/g) were present in higher concentrations. Likewise, in ethyl acetate fraction (St.Et.Ac), 21 phenolics were identified as 3,5-di-caffeoylquinic acid (177.4 mg/g) and 5-hydroxybenzoylquinic acid (46.9 mg/g) were most abundant phytochemicals. Highly valuable phenolics were also identified in other fractions including butanol (St.Bt), chloroform (St.Chf), and n-hexane (St.Hex). The various fractions exhibited concentration dependent inhibition of free radicals in DPPH and ABTS assays. Potent AChE inhibitory potentials were revealed by the test samples with St.Chf, St.Bt and St.EtAc being the most active having an IC50 of 298.1, 580.1, and 606.47 μg mL−1, respectively. Similarly, St.Chf, St.Bt, St.EtAc and St.Cr exhibited potent BChE inhibitory activity and was observed as 59.14, 54.73, 51.35 and 49.44%, respectively. A significant improvement in the exploratory behavior was observed in open field test and stress/anxiety was relieved effectively at 50-100 mg/kg. Likewise, EPM, light-dark and NOR tests revealed an anxiolytic and memory enhancing behaviors. These effects were further corroborated from the Y-maze and SWM transgenic studies that showed considerable improvement in cognition retention. ConclusionsThese findings concluded that S. tomentosa possessed potential anxiolytic and nootropic efficacies and may have therapeutic potential in neurodegenerative disorders.

Estimation of confidence intervals for quantitation of coeluted peaks in liquid chromatography–Photodiode array detection through a combination of multivariate curve resolution-alternating least-square and Bayesian inference techniques

There is a dramatic increase in drug candidates that exhibit complex structures and do not comply with Lipinski's rule of five. One of the most critical and complex technical challenges in the quality control of such drug candidates is the control of analogous substances contained in active pharmaceutical ingredients and related formulations. Although the development of ultrahigh-performance liquid chromatography and high-performance columns has improved efficiency per unit time, the difficulty of peak separation to quantify impurities with similar structures and physicochemical properties continues to rise, and so does the probability of failure to achieve the necessary separation. Coeluting peaks observed in the case of high-performance liquid chromatography (HPLC) with photodiode array detection can be separated using the multivariate curve resolution-alternating least-square (MCR-ALS) method exploiting differences in analyte UV spectra. However, relatively large quantitation errors have been observed for coeluting analogous substances, and the reliability of the corresponding quantitative data requires improvement. Herein, Bayesian inference is applied to separation by the MCR-ALS method to develop an algorithm assigning a confidence interval to the quantitative data of each analogous substance. The usefulness and limitations of this approach are tested using two analogs of telmisartan as models. For this test, a simulated two-component HPLC-UV dataset with an intensity ratio (relative to the main peak) of 0.1–1.0 and a resolution of 0.5–1.0 is used. The developed algorithm allows the prediction confidence interval, including the true value, to be assigned to the peak area in almost all cases, even when the intensity ratio, resolution, and signal-to-noise ratio are changed. Finally, the developed algorithm is also evaluated on a real HPLC-UV dataset to confirm that reasonable prediction confidence intervals including true values are assigned to peak areas. In addition to allowing the separation and quantitation of substances such as impurities challenging to separate by HPLC in a scientifically valid manner, which is impossible for conventional HPLC-UV detection, our method can assign confidence intervals to quantitative data. Therefore, the adopted approach is expected to resolve the issues associated with assessing impurities in the quality control of pharmaceuticals.

Trachystemon orientalis (L.) G. Don as a Valuable Source of Rosmarinic Acid: Biological Activities and HPLC Profiles.

Trachystemon orientalis (L.) G. Don, colloquially known in Türkiye as "kaldırık", is an edible plant belonging to the Boraginaceae. This plant has been practiced in traditional medicine for many years for its various therapeutic benefits. The effectiveness and chemical composition of plants can vary depending on their parts, age, and extraction solvent. Therefore, the current study aimed to define the biological activities of various parts and extracts of T. orientalis, which were collected in distinct seasons as young and mature, and investigate the main component responsible for these biological effects. Plant material was collected in different seasons from the northwest of Türkiye. 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activities were investigated to assess antiradical and antioxidant potential of the extracts. Anti-inflammatory activity of the extracts was also tested using human red blood cell membrane stabilizing method. Folin-Ciocalteu test was conducted to determine the total phenolic content. Reverse phase-high performance liquid chromatography-photodiode array detector (RP-HPLC-PDA) analysis was performed. Both methanol and aqueous extracts exhibited significant radical scavenging and anti-inflammatory activities compared with control (p<0.05). The highest percentage of inhibition on ABTS and DPPH free radicals was obtained in aqueous extracts of the mature herbs and roots, respectively. Methanol extracts of the mature roots and herbs exhibited the strongest anti-inflammatory capacity. Rosmarinic acid possessed a much higher antioxidant and anti-inflammatory effect than the reference compounds used in each assay in our study. High rosmarinic acid content of the extracts suggests that the compound responsible for the great biological activity potential is rosmarinic acid. To the best of our knowledge, the presence of rosmarinic acid in herbs and roots of T. orientalis was shown for the first time in our present study. Phytochemical composition and effective biological activities of T. orientalis explain its traditional use and indicate its significant potential in pharmaceutical industry applications.