Performance Liquid Chromatography-photodiode Array Research Articles

Holotrichones A and B, potent anti-leukemic lindenane-type sesquiterpene trimers with unprecedented complex carbon skeletons from a rare Chloranthus species

Two lindenane-type sesquiterpene (LDS) trimers with unprecedented carbon skeletons, holotrichones A (1) and B (2), were obtained from the whole plant of Chloranthus holostegius var. trichoneurus by a ultra performance liquid chromatography-photodiode array detector-mass spectrometry (UPLC-PDA-MS)-guided isolation strategy. Compound 1 represents the first LDS trimer incorporating a unique 3/5/6/6-fused framework, in which a lindenane-type monomer and the 2-methylbutyryl substituent of an LDS dimer is bridged by a six-membered ring system. Compound 2 is the first hetero-trimer fused by an LDS dimer with a p-benzoquinone-meroterpenoid, featuring an unusual 3/5/6/6/3/5/6/6/6 nonacyclic system fused by the sesquiterpenoid unit and a 2-geranyl-6-methyl-2,5-cyclohexadien-1,4-dione moiety. In compound 2, the dimeric LDS moiety is equipped with a rare oxaspiro[4.5]decane system. Their structures, including absolute configurations, were established by spectroscopic methods, GIAO NMR calculations and DP4+ probability analyses, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction analysis. The plausible biogenetic pathway speculation indicated that hetero- and homo-Diels-Alder additions may dominate the formation of these highly fused polycyclic frameworks. Both compounds 1 and 2 induced the human acute myeloid leukemia MV-4-11 cell death via apoptosis induction, which deserves further investigation on this new chemical class of LDS oligomers for their anti-leukemic potential.

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.

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.

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 &lt; 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.

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.

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.

A Fast and Robust Analytical Method Based on QuEChERS Technique using UPLC- PDA for Quantification of Carcinogenic Arylamines in Consumer Goods inclusive of Leather

Arylamines present in leather and textile products are reduced in vivo by cleavage of azo groups to form highly mutagenic and carcinogenic products that pose consumer’s health risk. The major textile and leather products that are offered to consumers need to be safe. In order to ensure safety as per the global regulations, the allowed limit of carcinogenic amines in textile and leather products cannot be more than 20 mg/kg. There are several methods that have been used for extracting the azo dyes and analyzing them in textile and leather products. Most of these methods are polluting from high solvent usage, time consuming, laborious, and have lower recovery rates. In the present study QuEChERS method, (dispersive solid phase extraction-dSPE) method was utilized for extraction of carcinogenic arylamines from dyed leather and textile products. The released arylamines are extracted into acetonitrile using QuEChERS salt and analyzed by Ultra Performance Liquid Chromatography-Photo Diode Array (UPLC-PDA). The extracted product was further confirmed by Gas Chromatography-Mass Spectrometer (GC-MS). The method proposed in this study is novel as it eliminates all significant concerns associated with the official ISO-17234-1 &amp;14362-1 test methods, such as use of solvents like tert-butyl methyl ether (MTBE) and the long duration of analysis. The sample preparation time is minimized from 120 min to 20 min. The developed method is easy to adopt with efficient recovery rate of arylamines in comparison to the official ISO method.

Detection of Carbamazepine, Diclofenac, and Ketoprofen via a SPE-HPLC-PDA Method in a Municipal Wastewater Treatment Plant in East Texas, USA

Increased demand for water reuse, recycling and reclamation heightens the need for improvements in the design of wastewater treatment plants that minimize or remove toxic pollutants or their metabolites. In this study, solid-phase extraction (SPE) technique utilizing C-18 cartridge was used for separation of three commonly used pharmaceuticals, namely, carbamazepine (CBZ), diclofenac (DF), and ketoprofen (KT) in wastewater samples. High performance liquid chromatography-photodiode array detector (HPLC-PDA) method was used for detection of CBZ, DF, and KT along the treatment stages (aeration, clarifier, raw activated sludge (RAS), chlorine contact chamber (CCC), and sulfur dioxide chamber) from Nacogdoches wastewater treatment plant (NWWTP), in East Texas, USA. Highest concentrations were found in the RAS followed by influent aeration stage. Pharmaceutical concentrations detected from the influent or RAS samples were 2.115 × 10−3 mg/L, 8.95 × 10−4 mg/L and 8.55 × 10−4 mg/L for CBZ, KT and DF, respectively. Highest detected total concentrations of pharmaceuticals were: carbamazepine (1.285 – 2.117 μg/L), diclofenac (0.600 – 0.865 μg/L), and ketoprofen (0.815 – 0.885 μg/L). The calculated % removal efficiencies along the treatment stages follow the order; CBZ: RAS > SO2 > clarifier ≈ aeration > CCC; KT: RAS > SO2 > aeration > CCC > clarifier; DF: RAS ≈ aeration ≈ CCC > clarifier > SO2. Determined removal efficiencies follow the order carbamazepine (∼ 39.2%) > diclofenac (∼ 30.6%) > ketoprofen (∼8.8%). Removal efficiencies of pharmaceuticals in the NWWTP may be affected by the human body, transition through activated wastewater treatment processes, and their metabolism and/or usage by humans.

Development and Validation of SI/RS-UHPLC-PDA Method for Olmesartan Medoxomil and Metoprolol Succinate-Related Substance.

Olmesartan medoxomil (OLM) and metoprolol succinate (MPS) in fixed-dose combination (FDC) tablet formulation prescribed extensively. Stability indicating (SI) method for impurities and related substance (RS) test quantitates the amount of these analytes in formulation; the manuscript presents SI/RS-ultra-high performance liquid chromatography-photodiode array (UHPLC-PDA) method for OLM and MPS and their impurities. Well-resolved separation of all analytes was achieved with gradient elution on a Shimadzu on Shimpack GIST-C18 (100 mm x 2.1 mm, 2 µm) column maintained at 25°C. Mobile phase-A consist of 0.1% orthophosphoric acid in water and mobile phase-B was acetonitrile at a flow rate of 0.4 mL/min, data integrated at 225 nm and 16 min of short runtime for satisfactory elution of all peaks. The proposed SI/RS-UHPLC-PDA method was developed and validated as per International Conference on Harmonisation (ICH) of Technical Requirements guidelines. The system suitability test complied by all eluted peaks of the interest with acceptable linearity, recovery, and precision. Specificity, robustness, and method sensitivity parameters were determined; all the parameters were found to be within the limits. All the impurities and stress-degraded peaks were well resolved. The proposed method was found to be simple, fast, linear, and accurate. Further, the method is precise, robust, and specific; suitable for routine IPQC during active pharmaceutical ingredient manufacturing, stability and impurity profiling studies of the titled bulk analytes. Furthermore, the method can be extended to assess the levels of impurities formed during life cycle of new FDCs of titled analytes.

Migration detection of six aromatic amines in polyamide food contact materials by HPLC after molecularly imprinted polymer pipette tip solid phase extraction

Some food contact materials (FCMs) may contain aromatic amines (AAs), which would migrate to food and pose a serious threat to human health. An extraction and determination method for AAs was established using pipette tip solid-phase extraction (PTSPE) combined with high performance liquid chromatography-photodiode array detector (HPLC-PAD). The PTSPE was based on molecularly imprinted polymer (MIP) monolith, which was synthesized in situ polymerization using benzidine as template molecule and 4-vinylpyridine as functional monomer. It was applied for selective extraction of six AAs from FCMs, including benzidine, 3,3′-dimethyl-4,4′-diaminodiphenylmethane, 2-methoxy-5-methylaniline, 3,3′-dimethoxybenzidine, 4,4′-methylenedianiline and 4-aminobiphenyl. The MIP monolith showed highly specific recognition and good adsorption ability for the six AAs, the imprinting factor for 4,4′-methylenedianiline can reach 4.29 and the adsorption capacity for 4-aminobiphenyl can reach 112.3 mg g−1. The linear range of the determination method for the six AAs was 0.60–500 µg kg−1 (R2＞0.997), the limits of quantitation was 0.60–4.50 µg kg−1 and the limits of detection (S/N = 3) was 0.20–1.50 µg kg−1. The MIP-PTSPE-HPLC-PAD method was applied for the migration detection of six AAs from polyamide kitchenware and the absolute recovery were 81.7–110.9%.

An In Vitro Study of the Healing Potential of Black Mulberry (Morus nigra L.) Extract in a Liposomal Formulation

Natural compounds are used in modern dermal treatments to avoid side effects commonly associated with conventional treatments. The aim of our study was to develop a liposomal formulation including black mulberry extract and to highlight its potential on the healing of normal human dermal fibroblasts (NHDF) in vitro using the scratch test. Mulberry-loaded liposomes (MnL) were prepared using a thin-film hydration method based on cholesterol (C) and phosphatidylcholine (PC) in a 1:3 (w/w) ratio. The liposomal formulation was characterized by analyzing its size, electric surface potential, morphology, entrapment efficiency, and in vitro healing effects. Also, the black mulberry fruits (Morus nigra L.) were characterized from point of view of polyphenolic compounds and antioxidant capacity by Ferric-Reducing Antioxidant Power (FRAP) assay. HPLC-DAD-MS (ESI+) (high performance liquid chromatography-photodiode array detection-mass spectrometry (electrospray ionization)) analysis indicated the presence of phenolic compounds namely from hydroxybenzoic and hydroxycinnamic acids and flavonols. Among flavonols, quercetin-glucoside represented 50.56%, and chlorogenic acid was the predominant compound among hydroxycinnamic acids (37.06%). In vitro fibroblast wound closure was more effective with mulberry-loaded liposomes (MnL) than extracts of mulberries. According to our study, mulberry-loaded liposomes have been shown to be effective in wound healing and can be used as a natural treatment.

Validated Bio-Analytical Method for Cinacalcet Concentrations in Rat Plasma by Reversed Phase-High Performance Liquid Chromatography- Photodiode Array Detector Method

The goal of this work was to create and verify a simple reversed phase-high performance liquid chromatography-photodiode array detector-based method for quantifying cinacalcet in rat plasma. Six albino Wistar rats were given cinacalcet orally by oral gavage. Blood was obtained at regular intervals using retro-orbital vein puncture and processed to get plasma. Cinacalcet was extracted from plasma using a 70:30 v/v ethanol:dichloromethane combination, with an 82.80 %-104.08 % recovery rate. The reversed phase-high performance liquid chromatography-photodiode array detector technique was used to determine cinacalcet plasma concentrations. The mobile phase was acetonitrile:0.1 M dipotassium phosphate buffer (pH 6.8) (50:50 v/v) at a flow rate of 1.0 ml/min. 241.0 nm was used as the detecting wavelength. The method's linearity for cinacalcet was proven by a correlation coefficient (R2) of more than 0.9996 and a linear range of 1 μg/ml to 24 μg/ml for cinacalcet. The lower and upper limits of quantification were found to be 1 μg/ml and 24 μg/ml respectively. At the cinacalcets retention time, no interfering peaks were seen. This reversed phase-high performance liquid chromatography-photodiode array detector approach is a straightforward and effective way to determine pharmacokinetic parameters by measuring plasma cinacalcet levels.

Valorisation of fruit peel bioactive into green synthesized silver nanoparticles to modify cellulose wrapper for shelf-life extension of packaged bread

Fruit peels are rich source of bioactive compounds such as polyphenols, flavonoids, and antioxidants but are often discarded as waste due to limited pharmaceutical and nutraceutical applications. This study aimed to valorise pomegranate and citrus fruit peel into green synthesised silver nanoparticles (AgNPs) in order to modify cellulose-based wrapping material for prospective food packaging applications and propose an alternate and sustainable approach to replace polyethene based food packaging material. Four different concentrations of AgNO3 (0.5 mM, 1 mM, 2 mM, and 3 mM) were used for green synthesis of AgNPs from fruit peel bioactive, which were characterised followed by phytochemical analysis. Ultraviolet–Visible spectroscopy showed surface plasmon resonance at 420 nm, XRD analysis showed 2θ peak at 27.8°, 32.16°, 38.5°, 44.31°, 46.09°, 54.76°, 57.47°, 64.61° and 77.50° corresponding to (210), (122), (111), (200), (231), (142), (241), (220) and (311) plane of face centred cubic crystal structure of AgNPs. Fourier-transform infrared spectroscopy analysis of AgNPs green synthesised from pomegranate and kinnow peel extract showed a major peak at 3277, 1640 and 1250–1020 1/cm while a small peak at 2786 1/cm was observed in case of pomegranate peel extract which was negligible in AgNPs synthesized from kinnow peel extract. Particle sizes of AgNPs showed no statistically significant variance with p > 0.10 and thus, 2 mM was chosen for further experimentation and modification of cellulose based packaging material as it showed smallest average particle size. Zeta potential was observed to be nearly neutral with a partial negative strength due to presence of various phenolic compounds such as presence of gallic acid which was confirmed by ultrahigh performance liquid chromatography-photodiode array(UHPLC-PDA) detector. Thermal stability analysis of green synthesised AgNPs qualified the sterilisation conditions up to 100 °C. AgNPs green synthesized from both the peel extracts had higher polyphenolic content, antioxidant and radical scavenging activity as compared to peel extracts without treatment (p < 0.05). The cellulose based food grade packaging material was enrobed by green synthesised AgNPs. The characterisation of modified cellulose wrappers showed no significant difference in thickness of modified cellulose wrappers as compared with untreated cellulose wrapper (p > 0.42) while weight and grammage increased significantly in modified cellulose wrapper (p < 0.05). The colour values on CIE scale (L*, a* and b*) showed statistically significant increase in yellow and green colour (p < 0.05) for modified cellulose wrappers as compared to control wrapper. The oxygen permeability coefficient, water vapour permeability coefficient, water absorption capacity and water behaviour characteristics (water content, swelling degree and solubility) showed significant decrease (p < 0.05) for modified cellulose wrapper as compared to control wrapper. A uniform distribution and density of green synthesised AgNPs across cellulose wrapper matrix was observed through scanning electron microscopy (SEM) images with no significant aggregation, confirming successful enrobing and stable immobilisation of nanoparticles from cellulose matrix. A seven-day storage study of bread wrapped in modified and control cellulose wrappers showed delayed occurrence of microbial, yeast and mould count in bread packaged in modified cellulose wrappers and thus, resulting in shelf life extension of bread. The results are encouraging for the potential applications of modified cellulose wrappers to replace polyethene based food packaging.

Analysis of seven selected antidepressant drugs in post–mortem samples using fabric phase sorptive extraction followed by high performance liquid chromatography-photodiode array detection

Fabric phase sorptive extraction (FPSE), a recently introduced microextraction technique, was herein applied for the first time to achieve a simple and rapid simultaneous extraction of seven common antidepressant drugs (ADs, venlafaxine, citalopram, paroxetine, fluoxetine, sertraline, amitriptyline, and clomipramine) in post–mortem samples, particularly whole blood and cerebrospinal liquor collected during autopsies. By eliminating the protein precipitation step and reducing solvent consumption, this technique resulted in sample preparation compliant with Green Analytical Chemistry (GAC) principles. FPSE uses a permeable and flexible substrate chemically coated with a sol–gel organic/inorganic sorbent as an extraction device. Among all tested FPSE membranes, the sol–gel Carbowax 20 M (sol–gel CW 20 M) coating on cellulose substrate showed optimal extraction efficiency for ADs. The selected drugs were analyzed and detected by the reverse phase high performance liquid chromatography (RP–HPLC) method coupled to photo diode array (PDA) detector. An isocratic elution that allows the complete separation of all analytes in only 20 min of chromatographic run time was applied using ammonium acetate buffer as aqueous mobile phase and acetonitrile (AcN) as the organic modifier. The limit of detection (LOD) ranged from 0.04 to 0.06 µg/mL, whereas limit of quantification (LOQ) was 0.1 µg/mL for all analytes except for Venlafaxine, which was 0.2 µg/mL.

Polyphenols in Different Plant Parts of Inula grandiflora Collected from Two Habitats of Uttarakhand Himalayas

ABSTRACT The phytoconstituent analyses have been undertaken for the first time from the different plant parts of Inula grandiflora Willd. collected from two habitats (open dry slopes and moist places) of Govind Pashu Vihar National Park and Wildlife Sanctuary, Uttarakhand, India. The total phenolic and flavonoid contents and antioxidant activity have been estimated from the methanolic extracts. Ten phenols and five flavonoids were quantified in the different parts using High Performance Liquid Chromatography-Photodiode Array Detection (HPLC-PDA) method. Among the plant parts, the trend of accumulation of total phenols and flavonoids was as follows: roots > flowers > stems > leaves in both populations. It has been observed that Kalkati Dhar germplasm has the highest phenolic content in the roots (16.42 ± 0.12 mg g−1 DW), while accession collected from Renugaad has the highest flavonoid content in the flowers (4.34 ± 0.024 mg g−1 DW). Eight bioactive compounds (caffeic acid, resorcinol, p-coumaric acid, ferulic acid, veratric acid, myristicin, coumarin, and quercetin) were found dominant in the Kalkati Dhar germplasm (3200 m asl, high altitude). Significantly, seven compounds, viz; vanillic acid, vanillin, cinnamic acid, epicatechin, catechol, luteolin, and rutin, were found predominate in the Renugaad germplasm (2750 m asl, low altitude). Overall, root part accumulates most of the bioactive polyphenolic compounds as compared to the aerial parts; therefore, it has the highest antioxidant potential.

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.

Fabric Phase Sorptive Extraction (FPSE) as an efficient sample preparation platform for the extraction of antidepressant drugs from biological fluids

• Fast method for simultaneous analysis of seven ADs. • Analysis of biological matrices. • FPSE application in conventional and unconventional matrices. • Method suitable for TDM of ADs. • Performances comparable to a more complex and expensive instrumentations. The quantification and interpretation of drug concentrations in biological matrices to optimize pharmacotherapy and to perform the therapeutic drug monitoring (TDM) is particularly important for compounds with narrow therapeutic ranges, known to cause adverse effects. In these cases, the biomonitoring is essential to avoid the toxicity and side effects. In this study, an innovative Fabric Phase Sorptive Extraction (FPSE) followed by high performance liquid chromatography-photodiode array detection (FPSE–HPLC–PDA) method was optimized and validated for the extraction and quantitative evaluation of seven antidepressant drugs (ADs, venlafaxine, citalopram, paroxetine, fluoxetine, sertraline, amitriptyline, and clomipramine) in human whole blood, urine, and saliva samples. The best chromatographic separation was obtained using a reverse phase column and ammonium acetate (50 mM, pH 5.5) and acetonitrile (AcN) as mobile phases, with 0.3% of triethylamine (TEA) for the best peak shape. The used sample preparation technique, FPSE, developed in 2014, has offered numerous advantages such as low consumption of organic solvents, no sample pretreatment, and reduced overall sample preparation time. Among all tested membranes, sol-gel carbowax (CW 20 M) sorbent, coated on cellulose FPSE media, was the most efficient. The developed method provides satisfactory limit of detection of 0.06 μg/mL for all analytes except for venlafaxine that was 0.04 μg/mL. Both RSD% and BIAS% gave values below ±15%, according to current guidelines. Finally, real samples analyzes were carried out, comparing the obtained data with the anamnestic data of the subjects, confirmed the validity of the method.