Performance Liquid Chromatography-photodiode Array Detection Research Articles

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.

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.

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.

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%.

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.

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.

Quantitative analysis of methoxyflavones discriminates between the two types of Kaempferia parviflora.

Kaempferia parviflora or black ginger is abundantly cultivated because its rhizomes contain methoxyflavones that have many pharmacological properties. K. parviflora can be divided into two types, based on morphological characteristics, but differences in their chemical compositions have never been explored. This research aims to find chemical markers that can be used to differentiate between the two types of K. parviflora, the red-leaf and green-leaf types, by quantifying the amounts of methoxyflavones. K. parviflora samples were collected from 39 locations in Thailand. Their genetic diversity was assessed by a genotyping-by-sequencing (GBS) technique to construct the population structure. Their chemical compositions were analyzed by high performance liquid chromatography-photodiode array detection to determine the methoxyflavone contents. The population structure based on >3,000 single nucleotide polymorphism (SNP) markers showed that the samples can be divided into two groups, which were consistent with the classification by leaf margin color (red-leaf and green-leaf types). HPLC analysis revealed 3,5,7,3',4'-pentamethoxyflavone (PMF), 5,7-dimethoxyflavone (DMF), 5,7,4'-trimethoxyflavone (TMF), 3,5,7-trimethoxyflavone and 3,5,7,4'-tetramethoxyflavone as major methoxyflavones that can be used as chemical markers. The red-leaf type showed higher amounts of PMF, TMF and 3,5,7,4'-tetramethoxyflavone than the green-leaf type, while the green-leaf type showed higher amounts of DMF and 3,5,7-trimethoxyflavone than the red-leaf type. These results provide another approach to discriminate the two types of K. parviflora using chemical profiles alongside genetic and morphological analyses. Therefore, a specific type of K. parviflora can be selected over the other based on preferences for a certain methoxyflavone.

Development and Validation of Ultra Performance Liquid Chromatography (UPLC) Method for the Quantitative Estimation of Caffeine in Non-Alcoholic Soft and Energy Drinks.

The popularity of beverages such as cold coffee, iced tea, and energy drinks has risen tremendously among athletes and youths. Energy drinks are used to enhance performance due to the presence of a high amount of caffeine (CFN) and sugars, as well as other constituents such as vitamins, amino acids, taurine, extracts of Ginkgo biloba, ginseng, guarana, and other herbal products. Commercial drinks are promoted as being beneficial to health; thus it is an important concern regarding adverse effects linked with these drinks or products. The aim of the study is to develop and validate the ultra performance liquid chromatography-photodiode array detector (UPLC-PDA) method for the estimation of CFN in eight marketed non-alcoholic drinks including two soft drinks and six energy drinks. The chromatographic separation of CFN was achieved on Acquity UPLC® CSHTM C18 1.7 µm, 2.1 × 100 mm column, using isocratic mode, mobile phase comprising acetonitrile and water (30:70, v/v) at a flow rate 0.3 mL/min using injection volume 1 µL. The elution of CFN occurred at 1.06 min, and the calibration curve of the CFN was computed from the peak area ratio detected at 273 nm. All the validation parameters were found within the assay variability limits as per ICH guidelines. The obtained results revealed that the soft drinks SD1 and SD2 have 87.21 ± 1.28 and 101.81 ± 1.52% (w/w), whereas CFN concentration in energy drink brands ED1, ED2, ED3, ED4, ED5, and ED6 was 95.90 ± 1.62, 64.0 ± 1.07, 76.68 ± 1.95, 74.97 ± 2.33, 82.09 ± 2.43, and 88.04 ± 2.94% (w/w) of labeled claims, respectively. The developed UPLC method was found suitable for the quality control of commercial soft and energy drinks containing CFN. The developed chromatographic method is very simple, cost effective and could be utilized for the routine analysis of caffeine in the soft and energy drinks.

UHPLC-MS Chemical Fingerprinting and Antioxidant, Antiproliferative, and Enzyme Inhibition Potential of Gaultheria pumila Berries.

Gaultheria pumila (Ericaceae) (known as Chaura or Mutilla) is a Chilean native small shrub that produces berry fruits consumed by local Mapuche people. In this study, the chemical fingerprinting and antioxidant, enzyme inhibition, and antiproliferative activities of the berries were investigated for the first time. Thirty-six metabolites were identified in the fruits by ultra-high performance liquid chromatography-photodiode array detection, hyphenated with Orbitrap mass spectrometry analysis (UHPLC-DAD-Orbitrap-MS). Metabolites, included anthocyanins, phenolic acids, flavonoids, iridoids, diterpenes, and fatty acids. Moderate inhibitory activities against acetylcholinesterase (7.7 ± 0.3 µg/mL), butyrylcholinesterase (34.5 ± 0.5 µg/mL), and tyrosinase (3.3 ± 0.2 µg/mL) enzymes were found. Moreover, selected major compounds were subjected to docking assays in light of their experimental inhibition. Results indicated that hydrogen bonding, π–π interaction, and a salt bridge interaction contributed significantly. Gaultheria pumila berries showed a total phenolic content of 189.2 ± 0.2 mg of gallic acid equivalents/g, total flavonoid content of 51.8 ± 0.1 mg quercetin equivalents/g, and total anthocyanin content of 47.3 ± 0.2 mg of cianydin-3-glucoside equivalents/g. Antioxidant activity was assessed using DPPH (92.8 ± 0.1 µg/mL), FRAP (134.1 ± 0.1 μmol Trolox equivalents/g), and ORAC (4251.6 ± 16.9 μmol Trolox equivalents/g) assays. Conversely, Gaultheria pumila showed a scarce antiproliferative potential against several solid human cancer cells. Our findings suggest that Gaultheria pumila berries have several bioactive metabolites with inhibitory effects against acetylcholinesterase, butyrylcholinesterase, and tyrosinase, and have the potential for use in food supplements.

Simultaneous determination of amitriptyline, nortriptyline, and clomipramine in aqueous samples using selective multi-template molecularly imprinted polymers

Molecularly imprinted polymers have gained attention as an efficient sorbent media to remove organic contaminants from diverse environmental matrices. In this research endeavor, multi-template molecularly imprinted polymer (mt-MIP) was synthesized and utilized in solid phase extraction (SPE) as a sorption medium for trace analysis of three tricyclic antidepressants (TCAs) (i.e. nortriptyline (NOR), amitriptyline (AMI), and clomipramine (CLO)) by high performance liquid chromatography-photodiode array detector (HPLC-PDA). Physicochemical properties of the synthesized sorbent were assessed by relevant spectroscopic and microscopic techniques to decipher their structural characteristics. mt-MIP with porous texture, cracks, and crevices on the external surface exhibit higher efficacy in extracting the target TCAs due to the enhanced tendency towards sorbate-sorbent interactions. The sorptive extraction of the mt-MIP was explored in standard and real samples (ground water, river water, hospital wastewater, and sewage water sample). mt-MIP integrated SPE technique coupled with HPLC-PDA exhibited linearity (R2 > 0.999) in the concentration ranging from 0.1 to 10 ng/mL with high precision (RSD% < 5) and accuracy (±3%). The developed method lower the detection limits for target TCAs to 0.016–0.029 ng/mL and provided good extraction efficiencies of 97.99 to 99.89%. The proposed mt-MIP-SPE-HPLC method offered high selectivity and sensitivity towards NOR, AMI, and CLO compared with the commercial C18-based SPE method. The spent mt-MIP was successfully regenerable (e.g., up to 8 cycles). The outcomes of the present work successfully revealed the apparent superiority of the mt-MIP towards the practical removal of target TCAs under real world conditions.

Simple HPLC method for rapid quantification of nicotine content in e-cigarettes liquids

AbstractElectronic nicotine delivery systems (ENDs) are gaining popularity in Jordan as alternatives to tobacco cigarettes with an estimation of 10% of tobacco smokers switching to ENDs. Since nicotine is toxic and highly addictive substance, it is important to develop and validate an easy and rapid analytical method to accurately measure nicotine level in e-liquids. A simple high performance liquid chromatography–photodiode array detection (HPLC–PDA) method was developed and validated for rapid determination of the actual nicotine content in 11 of the most popular e-liquids brands available in the Jordanian market and compared to the nicotine levels appeared in the labeled packaging. The new method of analysis showed an excellent linearity with correlation factor equal to 0.9994 with analytical range between 100 and 1,000 µg/mL, and Limit of detection (LOD) and Limit of quantification (LOQ) of 32.6 µg/mL and 98.9 µg/mL, respectively. The results showed that the actual measured nicotine concentrations ranged from 0 to 25.81 mg/mL with percent deviation ranged from 63.1% less than to 3.24% more than the labeled concentration on packaging. And more than 10% deviation difference in actual nicotine concentrations versus labeled were found in 9 of the 11 e-liquid products (82%). In conclusion, nicotine labelling among e-liquids products have not accurately reflect the actual content which may have potential negative impact on users.

A Facile Synthesized Polyaniline Coated Zerovalent Iron-Silica as an Efficient Sorbent for Magnetic Solid Phase Extraction of Phenolic Pollutants in Water Samples

A magnetic material based on polyaniline coated zerovalent iron-silica (Fe@SiO2@PANI) was simply synthesized using a facile manner under low energy consumption and applied for magnetic solid phase extraction of phenolic pollutants prior to analysis by high performance liquid chromatography-photodiode array detection (HPLC-PDA). The parameters which affected the extraction efficiency, i.e., sorbent amount, extraction time, sample volume and pH, desorption solvent and its volume, were optimized. Wide linear dynamic range of 0.012-10.000 μg mL-1 and the coefficients of determination better than 0.9926 were achieved. The limits of detection and the limits of quantitation were found in the ranges of 0.001-0.030 and 0.012-0.090 μg mL-1, respectively. The precision, expressed as relative standard deviation (RSD), was better than 11%. The synthesized sorbent exhibits good adsorption affinity and reusability up to 16 cycles. Applicability of the proposed methodology for analysis of phenolic residues in environmental and wastewater samples showed the recoveries in the range of 83.4-118.9%.

Separation and quantification of nine bioactive compounds in traditional Unani formulations by High Performance Liquid Chromatography–Photodiode Array Detector

Abstract A new High Performance Liquid Chromatography–Photodiode Array Detector (HPLC–PDA) method has been developed for the chromatographic separation and simultaneous quantitative determination of nine bioactive compounds, i.e. four phenolic (gallic acid, ellagic acid, chebulinic acid, and tannic acid), two flavanoids (rutin and quercetin), two anthraquinones (sennoside A and B) and one oxygenated hydrocarbon (vitamin C) in a well-known Unani polyherbal formulation namely Itrifal's. Separation was accomplished on a C18 LiChrospher 100 column (5 µm, 250 × 4.6 mm) with a gradient elution and recorded at 254 nm. The results demonstrated that the proposed method is reproducible, accurate, economic, and suitable for the quality control of traditional polyherbal Unani formulations containing complex compounds with different structures such as Itrifals.

A systematic investigation on free phenolic acids and flavonoids profiles of commonly consumed edible flowers in China

Edible flowers of ornamental plants are non-toxic flowers that have been used since ancient times for their nutritional and health benefits. These health benefits are attributed to their phytochemicals such as phenolic acids and flavonoids. These phytochemicals exert positive health effects on chronic diseases such as cancer, inflammation, and diabetes. This research paper is focused on the systematic investigation of free phenolic acids and flavonoids (including anthocyanins) present in 70 commonly consumed edible flowers collected from local parks and Qingping market of Guangzhou, China. The phenolic acids and flavonoids of flower samples were determined by high performance liquid chromatography-photodiode array detector. The phenolic acids, flavones, flavonols, flavanones, and anthocyanins of different flower samples exhibit a wide range of variation. Gallic acid (1177.8–27717.2 μg/g), protocatechuic acid (66.7–9641.2 μg/g), p-hydroxybenzoic acid (43.6–1792.7 μg/g), quercitrin and hesperidin (49.9–14576.6 μg/g), and quercetin and luteolin (8.8–480.0 μg/g) were the predominant phenolic compounds in edible flowers samples under investigation. Anthocyanins were detected only in 14 kinds of flower samples and most of these flowers were of red or purple colour.