High-performance Liquid Chromatography Photodiode Research Articles

In Vitro Analysis of Antimalarial Activity of Achillea fragrantissima (Forssk.) Sch.Bip Extracts Based on Beta-Hematin Formation

Introduction: The use of medicinal plants from traditional knowledge systems has increased in recent years. Achillea fragrantissima-infused water has a historical use in relieving gastrointestinal complaints and treating skin inflammations. With the rising resistance to antimalarial drugs, finding safe and affordable new drugs is crucial. Exploring medicinal plants shows promise for drug development in the fight against malaria. Methods: The objective of this research is to examine the in vitro antimalarial activity of A. fragrantissima-infused water, as well as water-ethanol extracts of its leaves, flowers, and stems based on β-hematin formation. Colorimetric semiquantitative assessments based on β-hematin inhibition and evaluation of optical density at 405 nm were employed. Results: The results demonstrated promising antimalarial activity of the different plant parts. The aqueous flower extract exhibited strong activity, comparable to that of the positive control. The leaves extract also showed strong antimalarial activity. The stem's aqueous and ethanolic extracts exhibited lower inhibitory activity compared to flowers and leaves. Synergistic antimalarial activity was observed when mixing the stem and leaves water extracts, as well as when mixing each of the stem and leaves extracts with the flower extract. The highest synergistic effect was observed when all three parts of the plant were mixed, resulting in activity comparable to the positive control. The high-performance liquid chromatography photodiode array (HPLC-PDA) chromatographic method confirmed the presence of rutin in the leaves extract. Conclusion: The findings of this study highlight the potential antimalarial activity of A. fragrantissima. The aqueous flower extract and leaves extract exhibited strong antimalarial activity, while the stem extracts showed relatively lower activity. Furthermore, synergistic effects were observed when combining different parts of the plant. These results suggest that A. fragrantissima holds promise as a potential source for the development of new antimalarial drugs. The confirmation of rutin presence in the leaves extract through HPLC-PDA analysis provides additional insights into the plant's chemical composition.

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.

Chemical composition, antioxidant properties, and antifungal activity of wild Origanum elongatum extracts against Phytophthora infestans.

Phytophthora infestans, the causative agent of late blight disease, has gained notoriety for its destructive potential, leading to substantial losses in potato yields. Although conventional systemic fungicides have been shown to be effective in controlling plant pathogens, growing environmental concerns have prompted the need for more integrated disease management approaches. Hence, in this study, the effectiveness of wild Origanum elongatum extracts as biopesticides was explored in controlling P. infestans and potentially mitigating its devastating impact in planta. The aerial parts of O. elongatum were subjected to sequential extraction using water, hexane, chloroform, and methanol. The obtained extracts were tested in vitro through the poisoned food procedure for their capacity to obstruct P. infestans growth and to defeat potato blight severity in vivo. The phyto-contents (total phenolic content (TPC) and total flavonoid content (TFC)), as well as the antioxidant activities, were spectrophotometrically determined in all extracts, and the phytoconstituents of the most active extract (methanolic extract) were profiled via high-performance liquid chromatography-photodiode array-tandem mass spectrometry (HPLC-PDA-MS/MS). In vitro, the complete inhibition rate of the P. infestans was obtained using the methanolic extract at 5 mg/mL, followed by the hexane and chloroform extracts at 10 mg/mL. Interestingly, complete inhibition of the pathogen was achieved upon the application of the aqueous extract at 10 mg/mL. In vivo, the aqueous extract at 25 mg/mL reduced the P. infestans severity rate to 27.25%, while the methanolic extract at 20 mg/mL led to the lowest severity rate. Moreover, the hexane and chloroform extracts impaired the pathogen severity rate to 50% and 41% using 20 mg/mL, respectively. The TPC and TFC in the extracts were variable with high concentrations detected in the methanolic extract with 485.42 mg GAE/g and 58.24 mg QE/g, respectively. In addition, the methanolic extract showed the highest antioxidant activities, while the chloroform extract exhibited the lowest activity. Liquid chromatography (LC)-MS/MS analysis of the methanol extract revealed 56 components from diverse classes. These included organic acids, phenolic acids, flavonoids, tannins, and coumarins. These findings suggest that O. elongatum could be investigated as a potential source of antifungal compounds targeting different phytopathogens.

Phenological Variations in the Content of Polyphenols and Triterpenoids in Epilobium angustifolium Herb Originating from Ukraine.

The composition of secondary metabolites undergoes significant changes in plants depending on the growth phase and the influence of environmental factors. Therefore, it is important to determine the harvesting time of plant material for the optimum secondary metabolite profile and therapeutic activity of the primary material. The shoots of Epilobium angustifolium are used as a healing tea due to the presence of polyphenolic compounds. The aim of this study was to assess the composition of phenolic compounds and triterpenoid saponins in E. angustifolium leaves and flowers and to estimate the dynamics of their content depending on the flowering phase. Qualitative and quantitative characterisation of polyphenols and triterpenoids in E. angustifolium samples from Ukraine of three flowering phases were performed using the high-performance liquid chromatography photo diode array (HPLC-PDA) method. During the present study, 13 polyphenolic compounds and seven triterpenoids were identified in the plant material. It was noted that the largest content and the best polyphenol profile was in late flowering. The most important polyphenolic compounds in the plant material were chlorogenic acid, hyperoside, isoquercitin, and oenothein B. The triterpenoid profile was at its maximum during mass flowering, with corosolic and ursolic acids being the dominant metabolites. The results of the analysis revealed that the quantity of many of the tested metabolites in the raw material of E. angustifolium is dependent on the plant organ and flowering phase. The largest content of most metabolites in the leaves was in late flowering. In the flowers, the quantity of the metabolites studied was more variable, but decreased during mass flowering and increased significantly again in late flowering. The results show that E. angustifolium raw material is a potential source of oenothein B and triterpenoids.

Novel stability indicating HPLC method for the quantification of Nirmatrelvir in bulk drugs

Nirmatrelviris anantiviral drug that acts asa 3C-likeprotease inhibitor. It is used with the combination of Ritonavir for the treatment of COVID-19. The World Health Organization strongly recommended this medicine to moderate and mild COVID patients, which is now the best option for COVID-19 therapeutics against SARS-CoV-2. As a result, there is an urgent need for quick and low-level quantitative analytical techniques for Nirmatrelvir and its products by focusing on the mass-compatible buffer with a shorter run time as the target. Accordingly, we developed a procedure with the following optimized conditions; mobile phase: 0.10 % formic acid in water and acetonitrile (55:45 %v/v), column oven temperature: 45 °C, flow rate: 1.0 mL/min with 15 min run time in isocratic elution mode. The developed procedure is linear and accurate from 0.20 to 500 µg/mL. Forced degradation studies in all solid and liquid degradations were executed to demonstrate the method's stability indicative capacity. To demonstrate the peak purity of the desired compound orthogonally, all degradation samples were then subjected to peak purity analysis using an HPLC PDA (High-performance liquid chromatography photodiode array) detector and LCMS (Liquid chromatography-mass spectrometry) analysis. The literature methods were developed with highly concentrated buffers which cause ion suppression in LCMS and increase backpressure in HPLC systems but the proposed method is conducive and more sensitive (limit of quantitation: 0.20 µg/mL and limitation of detection: 0.0 6 µg/mL) than the reported methods and has a wide linearity range as well. The developed technique has been fully validated by ICH (International Council for Harmonisation) guidelines. Further, it has been evaluated for greenness character using the AGREE (Analytical GREEnness) statistical tool and the obtained score is 0.61 out of 1.

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<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<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<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.

Identification of Acid Red 73 (CI 27290) in Cosmetic Hair Dye Preparations by High-Performance Liquid Chromatography – Photo Diode Array

Acid Red 73 (CI 27290) is a prohibited component in cosmetics, particularly in hair color formulations. The purpose of this investigation is to discover the coloring additive Acid Red 73 in cosmetic hair dye formulations. Acid Red 73 (CI 27290) is a sulfonated azo dye that is manufactured to be more hazardous than other colors and is damaging to the body. Based on variations in polarity and solubility, Acid Red 73 (CI 27290) is separated from the sample matrix and identified using High Performance Liquid Chromatography-Photo Diode Array (HPLC-PDA). The findings obtained from the tested samples satisfied the criteria since they did not include Acid Red 73 (CI 27290), as indicated by the difference in retention time and wavelength between the sample, the standard solution, and the spiked sample solution on the chromatogram.

Validasi Kurkumin Hasil Isolasi Rimpang Kunyit Dengan Kromatografi Cair Kinerja Tinggi Photodiode Array Detector

Curcumin is the main compound in turmeric and has problems in its stability. The analytical method used to determine curcumin levels is by using high performance liquid Chromatography (HPLC) Photodiode Array Detector (PDA). HPLC has a high separation capability and good reproducibility with a low detection limit. The purpose of this research is that the isolation curcumine determined by HPLC-PDA and has been validated can be applied as a standard in quantitative analysis in the laboratory. The validation parameters specified include accuracy, precision, linearity, detection limits and quantification limits. The results of HPLC-PDA research used acetonitrile mobile phase condition : buffer phosphate pH 3,5 (70:30 v/v), flow rate of 1 mL/min, wavelength 426 nm showed curcumin from isolation had resolution (Rs= 5.7172), linearity (r=0.9991), accuracy (0.5 and 1 ppm), presicion at a concentration of 5 ppm %RSD < 20.0%, and BD values also BK are 0.1122 and 0.3401 ppm. Validation of curcumin from isolation using HPLC-PDA has the requirements as a good validation and can be used as a reference in the laboratory.

Evaluation of the gastroprotective and ulcer healing properties by Fridericia chica (Bonpl.) L.G. Lohmann hydroethanolic extract of leaves.

Fridericia chica (Bonpl.) L.G. Lohmann (Bignoniaceae), is a climber native to Brazil, found in all Brazilian biomes. It is mostly known in Brazil as "carajiru," and home medicines made from the leaves have been used to cure disorders including stomach ulcers and other gastrointestinal disorders. The objective of the study was to investigate the F. chica hydroethanolic extract of leaves (HEFc) preventative and curative antiulcer gastrointestinal efficacy as well as the mechanisms of action using in vivo rodent models. F. chica was collected in the municipality of Juína, Mato Grosso, and its leaves were used to prepare the extract by maceration technique (70% hydroethanol in the 1:10 ratio, w/v) to obtain the HEFc. The chromatographic analysis of HEFc was carried out by High Performance Liquid Chromatography-Photo Diode Array-Electrospray Ionization-Mass Spectrometry (HPLC-PDA-ESI-MS)- LCQ Fleet™ system. To determine the potential antiulcer potential of HEFc (1, 5 and 20mg/kg, p.o.), the gastroprotective activity was assessed in various animal models of stomach ulcers caused by acidified ethanol, water constraint stress, indomethacin, (acute), and acid acetic (chronic). Additionally, the prokinetic properties of the HEFC were assessed in mice. The gastroprotective underlying mechanisms were evaluated by the histopathological analysis and determination of gastric secretion (volume, free and total acidity), gastric barrier mucus, activation of PGs, NO, K+ATP channels, α2-adrenoceptor, antioxidant activity (GSH, MPO and MDA), NO and mucosal cytokines (TNF-α, IL-1β, and IL-10) levels. The chemical composition of HEFc was analyzed and apigenin, scutellarin, and carajurone were identified. HEFc (1, 5 and 20mg/kg) showed effect against acute ulcers induced by HCl/EtOH with a reduction in the ulcerated area of 64.41% (p<0.001), 54.23% (p<0.01), 38.71% (p<0.01), respectively. In the indomethacin experiment, there was no change in the doses tested, whereas in the water immersion restraint stress ulcer there was a reduction of lesions at doses of 1, 5, and 20mg/kg by 80.34% (p<0.001), 68.46% (p<0.01) and 52.04% (p<0.01). HEFc increased the mucus production at doses of 1 and 20mg/kg in 28.14% (p<0.05) and 38.36% (p<0.01), respectively. In the pyloric ligation-induced model of gastric ulceration, the HEFc decreased the total acidity in all doses by 54.23%, 65.08%, and 44.40% (p<0.05) and gastric secretory volume in 38.47% at dose of 1mg/kg (p<0,05) and increased the free acidity at the dose of 5mg/kg by 11.86% (p<0.05). The administration of EHFc (1mg/kg) showed a gastroprotective effect possibly by stimulating the release of prostaglandins and activating K+ATP channels and α2-adrenoreceptors. Also, the gastroprotective effect of HEFc involved an increase in CAT and GSH activities, and a reduction in MPO activity and MDA levels. In the chronic gastric ulcer model, the HEFc (1, 5 and 20mg/kg) decreased the ulcerated area significantly (p<0.001) at all doses by 71.37%, 91.00%, and 93.46%, respectively. In the histological analysis, HEFc promoted the healing of gastric lesions by stimulating the formation of granulation tissue and consequently epithelialization. On the other hand, regarding the effect of HEFc on gastric emptying and intestinal transit, it was observed that the extract did not alter gastric emptying, but there was an increase in intestinal transit at the dose of 1mg/kg (p<0.01). These outcomes confirmed the advantages of Fridericia chica leaves for the treatment of stomach ulcers, which are well-known. HEFc was discovered to have antiulcer characteristics through multitarget pathways, which might be related to an increase in stomach defense mechanisms and a decrease in defensive factor. HEFc can be regarded as a potential new antiulcer herbal remedy because of its antiulcer properties, which may be attributed to the mixture of flavonoids, apigenin, scutellarin and carajurone.

Development and Validation of HPLC–Photodiode Array Method for Detecting Steroids in Skin Whitening Products Simultaneously

Since steroids are prohibited to be present in cosmetic products, thus an identification of these outlawed compounds intentionally incorporated in skin whitening product is enough for market surveillance purpose. Simultaneous method for detecting prednisone (PRN), triamcinolone acetonide (TCA), hydrocortisone acetate (HCA), clobetasole propionate (CBP), and mometasone furoate (MTF) has not been developed until now. The goals of this study were to develop and to validate for screening these five steroid compounds in cosmetic product by using reverse phase (RP) high-performance liquid chromatography (HPLC) equipped with photo diode array (PDA) detector. Validation method was applied including the system suitability test, determination of specificity and selectivity, and determination of detection limit. . In optimization step, we succeeded distinguish and identify five steroid compounds with different retention time.The retention times of PRN, TCA, HCA, CBP and MTF were detected at 3.267, 4.260, 4.910, 15.360, and 16.707 min, respectively. The method performed good selectivity with resolution more than 1.5. While for determination of limit of detection (LOD) was carried out by calibration curve approach over the range 0.5-5 µg/ml and resulted in LOD value for PRN, TCA, HCA, CBP and MTF were 7.20, 8.85, 8.17, 9.30, and 8.42 µg/g, respectively. The developed method can be applied for routine analysis of PRN, TCA, HCA, CBP and MTF in various type of cosmetic products.

Comparison of Different Green Extraction Techniques Used for the Extraction of Targeted Flavonoids from Edible Feijoa (Acca sellowiana (O.Berg) Burret) Flowers

This study aimed to investigate the effect of four green extraction techniques (ultrasound-assisted extraction, UAE; supercritical fluid extraction, SFE; subcritical water extraction, SWE; and extraction using deep eutectic solvents, DES) on the extraction of targeted flavonoids from edible feijoa flowers. The bioactive components in the obtained extracts were quantified by High-Performance Liquid Chromatography-Photodiode Array Detector (HPLC-PDA). Moreover, total polyphenol content and antioxidant activity by DPPH•, ABTS•+, FRAP, and CUPRAC assays were investigated. UAE generally gave the highest yields for isoquercitrin and quercetin content (18.36-25.33 and 10.86-16.13 µg/g), while DES extraction with choline chloride:lactic acid (1:2) and H2O content of 50% gave the highest yield of chrysanthemin (90.81 µg/g). The highest yield of flavone (12.69 mg/g) was obtained with supercritical CO2 at 300 bar. Finally, UAE gave the highest total polyphenol content (ca. 64 mg GAE/g) and antioxidant activity at 70 °C during 30 min with 40% (0.84 mmol TEAC/g and 2.25 mmol Fe2+/g, for ABTS•+ and CUPRAC, respectively) and 60% ethanol-water solution (0.49 mmol TEAC/g and 2.09 mmol Fe2+/g, for DPPH• and FRAP, respectively). The eco-friendly extraction techniques resulted in selective methods capable of extracting targeted bioactive compounds from edible feijoa flowers.

A new application of continuous sample drop flow microextraction using octanoic acid as a green extraction solvent for the determination of antibiotic drugs in urine samples.

In this study, octanoic acid (OA) was used as an extraction solvent for the pre-concentration and determination of three antibiotic drugs (levofloxacin, metronidazole, and tinidazole) in urine samples. To extract the antibiotic drugs, a green solvent was used as the extraction solvent in the continuous sample drop flow microextraction method, followed by a high-performance liquid chromatography photodiode array detector. According to the findings, the present study offers an environmentally friendly analytical method with a high capacity for the microextraction of the antibiotic drugs at very low concentrations. The calculated detection limits were 6.0-10.0µg/L and the linear range was found between 20 and 780µg/L. The proposed method showed excellent repeatability with the RSD values ranging from 2.8 to 5.5%. The relative recoveries were between 79.0 and 92.0% in the urine samples with spiked levels of 40.0-100.0µg/L for metronidazole and tinidazole, and 100.0-200.0μg/L for levofloxacin.

Effect-Directed, Chemical and Taxonomic Profiling of Peppermint Proprietary Varieties and Corresponding Leaf Extracts

During the development of novel, standardized peppermint extracts targeting functional applications, it is critical to adequately characterize raw material plant sources to assure quality and consistency of the end-product. This study aimed to characterize existing and proprietary, newly bred varieties of peppermint and their corresponding aqueous extract products. Taxonomy was confirmed through genetic authenticity assessment. Non-target effect-directed profiling was developed using high-performance thin-layer chromatography-multi-imaging-effect-directed assays (HPTLC-UV/Vis/FLD-EDA). Results demonstrated substantial differences in compounds associated with functional attributes, notably antioxidant potential, between the peppermint samples. Further chemical analysis by high-performance liquid chromatography-photodiode array/mass spectrometry detection (HPLC-PDA/MS) and headspace solid-phase microextraction-gas chromatography-flame ionization/MS detection (headspace SPME-GC-FID/MS) confirmed compositional differences. A broad variability in the contents of flavonoids and volatiles was observed. The peppermint samples were further screened for their antioxidant potential using the Caenorhabditis elegans model, and the results indicated concordance with observed content differences of the identified functional compounds. These results documented variability among raw materials of peppermint leaves, which can yield highly variable extract products that may result in differing effects on functional targets in vivo. Hence, product standardization via effect-directed profiles is proposed as an appropriate tool.

Chemical Profiling of Citrus sinensis Root and the Effects of Its Secondary Metabolites on Cisplatin-Induced Renal and Cardiac Toxicities

Citrus fruits are among the most important economical crops, because of their nutritional value, medicinal importance, and unique flavor. Gas liquid chromatography-mass spectrometry of the hydro-distilled oil from the root resulted in the identification of 110 compounds with germacrene B (22%), aromadendrene (21.6%), α-santalene (7.1%), geijerene (4,81%), germacrene D (4.3%), and limonene (3.4%) as major constituents. In addition, chemical profiling the dichloromethane fraction of the root analyzed by high-performance liquid chromatography-photo diode array detector-electrospray tandem mass spectrometry afforded the identification of 43 compounds belonging to acridone alkaloids, coumarins, and flavonoids. Moreover, xanthyletin, citracridone I and II, clausarin, O-methylcitrusinine-I, and grandisinine were isolated as major metabolites using column chromatography and characterized depending on different spectroscopic techniques. Xanthyletin and citracridone I were investigated for their in vitro cytotoxicity against hepatocellular carcinoma and breast adenocarcinoma cell lines, and in vivo protective effect against cisplatin-induced nephrotoxicity and cardiotoxicity in different dose levels in a rat model. Xanthyletin and citracridone I showed protective activity against cisplatin-induced nephrotoxicity. It attenuated cisplatin-induced elevation of both serum urea and creatinine in a dose-dependent manner. Moreover, xanthyletin attenuated cisplatin-induced elevation of malondialdehyde and glutathione in both renal and cardiac tissues.Graphical abstract

Separation and Detection of Abamectin, Ivermectin, Albendazole and Three Metabolites in Eggs Using Reversed-Phase HPLC Coupled with a Photo Diode Array Detector

An innovative and sensitive approach using high-performance liquid chromatography-photo diode array detection (HPLC-PDAD) was developed and optimized for the simultaneous determination of abamectin (ABM), ivermectin (IVM), albendazole (ABZ) and three metabolites in eggs. The samples were extracted with acetonitrile (MeCN)/water (90:10, v/v), and the extracts containing the targets were cleaned up and concentrated by a series of liquid-liquid extraction (LLE) steps. A reversed-phase C18 column and a mobile phase consisting of 0.1% trifluoroacetic acid (TFA) aqueous solution and methanol (MeOH) were utilized to perform optimal chromatographic separation. The developed method was validated on the basis of international guidelines. The limits of detection (LODs) and quantitation (LOQs) were 2.1-10.5 µg/kg and 7.8-28.4 µg/kg, respectively. Satisfactory linear relationships were observed for the targets in their corresponding concentration ranges. The mean recoveries ranged from 85.7% to 97.21% at 4 addition levels, with intraday and interday relative standard deviations (RSDs) in the ranges of 1.68-4.77% and 1.74-5.31%, respectively. The presented protocol was demonstrated to be applicable and reliable by being applied for the detection of target residues in locally sourced egg samples.

Furaneol Detection with Molecularly Imprinted Polymer (MIP)-Based Polyaniline (PANI) Gas Sensor

Strawberry is one of the beloved fruits worldwide due to its exquisite flavor with highly nutritious compounds which give promising health benefits. [1] With dragging the great attention, efforts to monitor the quality of strawberries and strawberry derivatives are becoming significant to obtain better flavor and nutrition. 2,5-Dimethyl-4-hydroxy-3(2H)-furanone(furaneol) is one of the compounds which plays an important role in a flavor that can be naturally found in various fruits such as strawberry, pineapple, raspberry, and mango. Especially, furaneol composition is reported as a significant factor for maturity condition monitoring of strawberries due to the different amounts at the ripening stage. [2] Furaneol is also known to be an essential signature for controlling the food production process, verifying the food production origin, and regulating the quality of products.Currently, furaneol detection is conducted with time-consuming techniques, such as gas chromatography (GC) and high-performance liquid chromatography (HPLC). For example, Yuan et al. [3] showed the identification of furaneol with HPLC. Also, Buttery et al. [4], and Lopez et al. [5], used GC for the determination of furaneol in foods. Developing a real-time gas sensor for fast, cheap, stable, sensitive detection of furaneol is a significant challenge in the area of the food industry.Chemo-resistive gas sensors can be an attractive approach due to their miniaturization, simple operation, ease of fabrication, and low production cost. For utilizing resistive gas sensors, polyaniline (PANI) got great attention because of its environmental stability, simple synthesis, and ease of tailoring the surface charge characteristics by changing the dopants. However, PANI-based gas sensors are reactive to numerous gases which limits the selective detection of target gas molecules. Many compounds in strawberries may restrict the use of PANI-based sensors. [6] Molecular imprinting technology (MIT), which can create molecularly imprinted polymers (MIPs) obtaining specific binding sites for target molecules, is utilized to achieve selective gas sensing similar to biological antibodies. [7] The improved affinity not only provides selective detection but also enhances response to target gas molecules.In this study, a resistive gas sensor based on synthesized molecularly imprinted polymer and polyaniline is demonstrated for the detection of furaneol gas molecules for its operation in food packaging. Herein, we developed a unique method to synthesize molecularly imprinted polymer and polyaniline (MIP-PANI) nanocomposites with an interfacial polymerization technique. Aniline monomer is polymerized and oxidized simultaneously to tailor the surface charge characteristics forming polyaniline. Then, the molecularly imprinting process on polyaniline is conducted with methacrylic acid (MAA) as a functional monomer, ethylene glycol dimethacrylate (EGDMA) as a crosslinker, furaneol as a template, and benzoyl peroxide as a radical initiator. After the furaneol template was constructed, MIP-PANI presented a high affinity to furaneol target molecules and showed a high sensing response. Their response signals are enhanced tens of times compared to the pure PANI. Furthermore, selective detection of the target furaneol molecules was observed. This study may broaden the application of resistive gas sensors for selective and sensitive detection of volatile organic compounds in food industries. Reference [1] S. Afrin, M. Gasparrini, T.Y. Forbes-Hernandez, P. Reboredo-Rodriguez, B. Mezzetti, A. Varela-Lopez, F. Giampieri, M. Battino, Promising Health Benefits of the Strawberry: A Focus on Clinical Studies, J Agric Food Chem 64 (2016) 4435-4449. [2] C. Aubert, S. Baumann, H. Arguel, Optimization of the analysis of flavor volatile compounds by liquid-liquid microextraction (LLME). Application to the aroma analysis of melons, peaches, grapes, strawberries, and tomatoes, Journal of Agricultural and Food Chemistry 53 (2005) 8881-8895. [3] J.P. Yuan, F. Chen, Separation and identification of furanic compounds in fruit juices and drinks by high-performance liquid chromatography photodiode array detection, Journal of Agricultural and Food Chemistry 46 (1998) 1286-1291. [4] R.G. Buttery, G.R. Takeoka, M. Naim, H. Rabinowitch, Y. Nam, Analysis of furaneol in tomato using dynamic headspace sampling with sodium sulfate, Journal of Agricultural and Food Chemistry 49 (2001) 4349-4351. [5] R. Lopez, M. Aznar, J. Cacho, V. Ferreira, Determination of minor and trace volatile compounds in wine by solid-phase extraction and gas chromatography with mass spectrometric detection, Journal of Chromatography A 966 (2002) 167-177. [6] A.T. Oz, G. Baktemur, S.P. Kargi, E. Kafkas, Volatile Compounds of Strawberry Varieties, Chemistry of Natural Compounds 52 (2016) 507-509. [7] L. Chen, X. Wang, W. Lu, X. Wu, J. Li, Molecular imprinting: perspectives and applications, Chem Soc Rev 45 (2016) 2137-2211.

Commelina benghalensis (Wandering Jew) Linn exhibits abortifacient potentials and hepatotoxicity in pregnant Wistar rats via elevating indicators of oxidative stress and activating proinflammatory cytokines

Ethnopharmacological relevanceCommelina benghalensis Linn is a perennial plant with upright stems reaching a height of 1 m. Its stem is commonly used to induce abortion in traditional medicine. However, there are insignificant scientific data to evaluate such a claim. Aim of the studyThe study was conducted to determine the abortifacient and toxicological potential of ethanol extract of Commelina benghalensis Linn stem (EECBS) via selected proinflammatory and oxidative stress biomarkers in pregnant Wistar rats. Materials and methodsTo determine the phytochemicals responsible for EECBS's toxicity and abortifacient effects, high-performance liquid chromatography–photodiode array detection (HPLC-PDA) and gas chromatography-mass spectrometry (GC-MS) was used. The abortion rate was determined by monitoring the markers of reproductive system failure in the experimental model. To assess rat hepatotoxicity, biochemical markers and immunohistopathological parameters were used. ResultsResults demonstrated the presence of isomeric benzene-mesitylene compounds in EECBS. Also, EECBS significantly altered the markers of liver function and oxidative damage while eliciting a significantly reduced (P < 0.05) number of live fetuses, number of corpora lutea, progesterone, estradiol, and luteinizing hormone, whereas the number of dead fetuses percentage vaginal opening, and post-implantation loss increased significantly (P < 0.05). Estrogenicity studies indicated a significant (P < 0.05) increase in uterine weight, uterine glucose, and ALP dose-dependently. Moreover, EECBS also caused a vaginal hemorrhage preceding the parturition. Also, EECBS treatment significantly increased levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and significantly elevated the expression of COX-2 protein in the liver. ConclusionThe current investigation established Commelina benghalensis Linn stem's abortifacient activity. Continuous use, on the other hand, may cause liver damage in pregnant rats by disrupting antioxidant defense mechanisms, promoting the production of pro-inflammatory cytokines, and increasing COX-2 expression. Hence, caution should be excised while consuming this plant's stem for medication purposes, especially during the gestational period.

A Validated Stability-Indicating HPLC-PDA Method for Tolnaftate: Identification, Characterization and In Silico Toxicity Predictions of Major Degradation Products.

Simple and rapid stability indicating High Performance Liquid Chromatography-Photo Diode Array (HPLC-PDA) method was developed and validated for the estimation of tolnaftate in the presence of its forced degradation products. The method employed SunQSil C18 column (250mm × 4.6mm, 5μm) as stationary phase and acetonitrile:water (85:15, v/v) as mobile phase. Retention time of tolnaftate was 6.9min. Acid and alkali hydrolysis, oxidation, photo degradation and thermal degradation studies were carried out to evaluate the degradation behavior of tolnaftate. The developed and optimized method was validated as per International Conferences on Harmonization (ICH) guidelines. Limit of detection and limit of quantitation were found to be 0.092 and 0.276μg/mL, respectively. Linearity was observed in a concentration range of 0.276-6μg/mL with R2= 0.9936. %Recovery was found to be between 98.28% and 100.71%. The developed and validated Reversed Phase-High Performance Liquid Chromatography (RP-HPLC) method was successfully applied for quantification of tolnaftate in in-house topical solution. Major base and oxidative degradation products were identified and characterized by liquid chromatography-electrospray ionization mass spectrometry. The probable mechanisms for the formation of degradation products were predicted based on the fragmentation pattern of degradation products. The in silico dermal penetration predictions and carcinogenicity of degradation products were evaluated by using QikProp and CarcinoPred-EL functionality.

METHOD DEVELOPMENT AND VALIDATION OF CEFOPERAZONE AND SULBACTAM IN DRIED BLOOD SPOTS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY PHOTODIODE ARRAY DETECTOR

Objective: The primary purpose of this research was to develop a simple, precise, fast, and accurate method for measuring cefoperazone and sulbactam simultaneously in dried blood spots (DBS) using HPLC PDA. Methods: A simplified analytical method for quantifying cefoperazone and sulbactam in DBS samples using a High-Performance Liquid Chromatography photodiode array detector with isocratic elution was developed and validated. The best chromatographic conditions were obtained by using a reversed-phase column (250 x 4.6 mm; 5 mm); phosphate buffer 10 mmol pH 3.2–acetonitrile (83:17, v/v) as a mobile phase; a flow rate of 1.0 ml/min; a column temperature of 35 °C; a photodiode array detector at 210 nm, and cefuroxime as internal standard. Samples were prepared by liquid-liquid extraction with 100 ml hydrochloric acid 0.5 mol/l and 1000 ml ethyl acetate, evaporated with nitrogen and reconstituted with 100 mL phosphate buffer–acetonitrile (4:1). Results: The total chromatography run time was 15 min, and the elution times for sulbactam, cefoperazone, and IS (cefuroxime) were 3.46, 10.221, and 6.987 min, respectively. A linear response function was established at 0.5-30 mg/ml with (r) 0.995 for sulbactam and 2.5-250 mg/ml with (r) 0.999 for cefoperazone in dried blood spots. The lower limit quantification (LLOQ) concentration of sulbactam 1 mg/ml and cefoperazone were 5 mg/ml. Conclusion: This method has successfully fulfilled the validation requirement referring to the 2011 EMA and 2018 FDA guidelines.

Prunus lusitanica L. Fruits as a Novel Source of Bioactive Compounds with Antioxidant Potential: Exploring the Unknown.

Prunus lusitanica L., also known as Portuguese laurel or locally known as ‘azereiro’, is a rare species with ornamental and ecological value. Only two studies regarding the bioactivity and chemical composition of its leaves were reported to date. Thus, the present study aims to qualitatively and quantitatively evaluate the phenolic profile, through HPLC-PAD-ESI-MS/MS (high-performance liquid chromatography–photodiode array detection–electrospray ionization tandem mass spectrometry), as well as the radical scavenging capacity, through ABTS (2,2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) and DPPH (2,2-diphenyl-1 picrylhydrazyl), and the reducing power (FRAP, ferric reducing antioxidant power) assays, of P. lusitanica fruits during a 4-year study. In total, 28 compounds were identified and quantified in the fruits, including 21 hydroxycinnamic acids (60.3%); 2 flavan-3-ols (27.9%), 2 anthocyanins (10.5%), 2 flavonols (1.0%), and 1 secoiridoid (0.3%). High antioxidant capacity was observed, with ABTS values ranging from 7.88 to 10.69 mmol TE (Trolox equivalents)/100 g fw (fresh weight), DPPH values from 5.18 to 8.17 mmol TE/100 g fw, and FRAP values from 8.76 to 11.76 mmol TE/100 g fw. According to these results, it can be concluded that these are rich sources of phenolic compounds with very promising antioxidant capacity and, therefore, with potential applications in the food and/or phytopharmaceutical sectors.