Objective: The aim of this research was to develop and validate identification and Quantitation methods for 1,4-naphthoquinone in the extract of Eleutherine bulbosa.
Methods: High-performance liquid ...chromatography (HPLC) and high-performance thin-layer chromatography with densitometric detection (HPTLC-densitometry) were employed as analytical techniques. HPTLC-densitometry was performed at a wavelength of 249 nm, while UHPLC was conducted at a wavelength of 254 nm. Both methods were utilized to analyze 1,4-naphthoquinone in 96% ethanol extract of E. bulbosa as a Quantitation parameter in the standardization of the formulation. HPTLC separation was carried out on a 20 cm × 20 cm HPTLC glass plate coated with silica gel 60 F254 using a mobile phase of chloroform: methanol (8:2, v/v). For HPLC analysis, a C18 column with an isocratic method was employed using a mobile phase of 95% methanol in pump A and 0.5% chloroform in pump B. The calibration curve of peak area against concentration showed linearity within the range of 2500-15000 ppm/spot−1 and 3–21 μg/ml for HPTLC and HPLC, respectively. Both methods were validated for accuracy, precision, linearity, selectivity, LOD, and LOQ.
Results: The results showed that both methods exhibited linearity that met the standards, as they produced correlation coefficients (r) greater than 0.9900. Furthermore, both methods demonstrated good accuracy, with consecutive recovery values of 99.53% and 101.89%. On the other hand, the methods fulfilled the precision requirements, with respective values of 0.7159% and 2.884% (in compliance with the requirement of<5%). Additionally, to meet the LOD and LOQ requirements in HPTLC, the LOD value obtained was 163 ppm, and the LOQ value was 495 ppm. In HPLC, the retention time of the standard 1,4-naphthoquinone and the analyte compound in the extract of E. bulbosa were the same, at 3.507 min. The selectivity test on HPTLC indicated that the 1,4-naphthoquinone compound was at an RF value of 0.81, which was also detected in the extract of E. bulbosa at the same RF value.
Conclusion: In conclusion, our findings demonstrate that HPLC and HPTLC methods for the determination of 1,4-naphthoquinone content have met the standards for linearity, accuracy, precision, selectivity, LOD, and LOQ. Therefore, these methods can be recommended for the quality control of raw materials of E. bulbosa extract.
•Advanced glycation end products (AGEs) can be formed by Maillard reactions.•High-fat chips and crackers contained high amounts of AGE precursors.•Low fat containing breakfast cereals had low amounts ...of AGE precursors.•High salt content in crackers may affect the increased amount of glyoxal and methylglyoxal.
Consumption of processed foods is increasing in today's modern diet. Advanced glycation end products (AGEs) can be formed by Maillard reactions as well as oxidation of proteins and fats in food processing. The aim of the present study was to determine the amount of glyoxal and methylglyoxal in chips, crackers, and breakfast cereals and to evaluate their effects on human health. In this research, chips (26), crackers (5), and breakfast cereals (11) were obtained from different markets in Istanbul, Turkey. The amounts of glyoxal and methylglyoxal in these foods were determined by high performance liquid chromatography (HPLC) using 4-nitro-1,2-phenlenediamine as a pre-column derivatizing reagent. The measured amount of glyoxal and methylglyoxal ranged between 94 and 1464 and 123–661 µg/100 g in chips, and between 338 and 1936 and 727–1397 µg/100 g in crackers, and between 8 and 1575 and 111–1201 µg/100 g in breakfast cereals, respectively. The products used in this study, especially chips and crackers, contained high fat and were baked at a high cooking temperature. Therefore, these products had higher amounts of AGE precursors. Besides, high salt content in crackers may affect the increased amount of glyoxal and methylglyoxal. On the contrary, low fat containing breakfast cereals had lower amounts of AGE precursors. People who often consume AGE-rich snack foods will be at higher health risk than those who consume less. The health problems associated with AGEs can be reduced with an AGE-restricted diet.
Comparison of antioxidant, anti-inflammatory and xanthine oxidase inhibitory activities of extracts from ginger (extractswith different polarity solvents) and its 6-gingerol, 6-shogaol, and 6-paradol ...chemical constituent’s in vitro systems were examined. The highest scavenging activity of ginger and its active constituents was found against H2O2 and FRAP as compared to DPPH assay and the strongest enzyme inhibition found against lipoxidase, xanthine oxidase, β-glucuronidase, and hyaluronidase both by ginger extracts and its active constituents. The extraction and analysis of ginger extracts by chromatographic methods like HPTLC and HPLC revealed the presence of the principal substances like 6-gingerol, 6-shogaol, and 6-paradol which are responsible for the biological activities.
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Ginger, Zingiber officinale Roscoe, is a spice used as a medicinal plant in many countries. We are the first to report the HPTLC analysis of ginger extract and analysis of their active principles with comparative antioxidant, anti-inflammatory, and xanthine oxidase inhibitory activities. The five fractions were obtained by using different polarity solvents with selective extraction procedure from ginger rhizomes and found that they revealed the difference in bioactivity against studied parameters. The ethyl acetate extract (EAE) showed significant antioxidant activity studied by DPPH, FRAP, and H2O2 assay (IC50±SEM μg/mL: 6.8±0.6, 12±0.2, and 20±2.5, respectively). In the xanthine/xanthine oxidase system, the antioxidant potentials of EAE and the water extract (WE) (% inhibition: 76% and 74%, respectively) were higher than those of the ethanol extract (EE), diethyl ether extract (DEE), and n-butanol extract (NBE). Regarding anti-inflammatory activity, EAE exhibited greater inhibition of lipoxidase (80%), and β-glucuronidase (78%) compared to hyaluronidase (46%) and diene-conjugates (37%). Chromatographic analysis revealed that several principal substances including 6-gingerol, 6-shogaol, and 6-paradol were responsible for the biological activities for ginger. Compound 6-gingerol revealed high FRAP-reducing activity (IC50±SEM μM: 5±0.4). 6-Gingerol also significantly inhibited the activities of xanthine oxidase (85%), lipoxidase (87%), β-glucuronidase (85%), and hyaluronidase (56%), respectively. These results indicated that ginger rhizome fractions and its active constituents having promising antioxidant, anti-inflammatory, and anti-gout properties and might be used as potential natural drug against oxidative stress and inflammatory related diseases after successful in vivo study and clinical trials.
Sodium benzoates as a chemical preservative are common practice in modern food technology. It is permitted by international laws in food as a food additive with a restricted amount. But it is highly ...used in different food products like fruit juices. Therefore, an experimental study was conducted to determine the concentration of sodium benzoate in different brands of orange juices available in the market Tangail region, Bangladesh using High Performance Liquid Chromatography (HPLC). Chromatographic analysis was performed by a column (250 × 4.6 mm) with an isocratic solvent system at 40°C. The mobile phase consisted of sodium acetate and acetic acid buffer (pH was 4). The correlation of coefficient was 0.9999 from the standards curves and the variety of external standard absorptions was 0.93 to 20.05 µg/mL. Analysis of four brands of orange juice samples showed that the concentration of sodium benzoate was within the FDA standard, but two brand samples slightly exceeded the Bangladesh Standard and Testing Institute (BSTI) standard. Brand 1 and Brand 4 samples contained sodium benzoate 14.61 and 9.59 mg/100 mL respectively which were within the level BSTI standard range whereas Brand 2 and Brand 3 samples contained 16.11 and 16.33 mg/100 mL respectively which exceeded the level of BSTI standard.
•This work proposes a strategy for the use of a waste material from plum.•An extract with a 35% of proteins (in dried and defatted plum seeds) was obtained.•Peptides with antioxidant capacity were ...obtained from these proteins.•Peptides with ACE inhibiting capability were obtained from these proteins.•Identified peptides showed features common in antioxidant and ACE inhibiting peptides.
Fruit processing produces a great amount of wastes. Stone of fruits such as plum (Prunus Domestica L.) are part of these residues. Plum stones contain a seed with a high content in proteins and lipids that are mostly underused and undervalued. This work proposes to extract proteins from this by-product, to evaluate their potential as bioactive peptide source, and to identify these peptides using RP-HPLC–MS/MS. A method for the extraction of plum seed proteins using high intensity focused ultrasounds has been developed. Extracted proteins have been digested using Alcalase, Thermolysin, Flavourzyme, and Protease P enzymes. Alcalase was the enzyme yielding the extracts with the highest ABTS radical scavenging capacity and lipid peroxidation inhibition capacity. It also showed a high capacity to scavenge hydroxyl radicals and to inhibit ACE enzyme. Analysis of the Alcalase extract by RP-HPLC–ESI–Q-TOF enabled the identification of short chain and highly hydrophobic peptides.
•S-adenosylmethionine and S-adenosylhomocysteine have been separated using HPLC strategies.•RP-ESI-MS/MS provided best analytical figures of merit for the purpose.•The methodology has been applied to ...cancer cells sensitive and resistant to cisplatin.•Positive correlation of SAM/SAH ratio with the methylation of DNA has been found.
S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are essential compounds in the carbon metabolic cycle that have clinical implications in a broad range of disease conditions. The measurement of the ratio SAM/SAH also called methylation index, has become a way of monitoring the DNA methylation of a cell which is an epigenetic event with important clinical implications in diagnosis; therefore the development of suitable methods to accurately quantify these compounds is mandatory. This work illustrates the comparison of three independent methods for the determination of the methylation index, all of them based on the chromatographic separation of the two species (SAM and SAH) using either ion-pairing reversed phase or cation exchange chromatography. The species detection was conducted using either molecular absorption spectrophotometry (HPLC–UV) or mass spectrometry with electrospray (ESI-MS/MS) as ionization source or inductively coupled plasma (DF-ICP-MS) by monitoring the S-atom contained in both analytes. The analytical performance characteristics of the three methods were critically compared obtaining best features for the combination of reversed phase HPLC with ESI-MS in the MRM mode. In this case, detection limits of about 0.5ngmL−1 for both targeted analytes permitted the application of the designed strategy to evaluate the effect of cisplatin on the changes of the methylation index among epithelial ovarian cancer cell lines sensitive (A2780) and resistant (A2780CIS) to this drug after exposition to cisplatin.
Hydrophilic interaction chromatography (HILIC) works with organic solvent-water mixtures as eluent and is based on the formation of a water enriched liquid phase on the surface of a hydrophilic ...stationary phase. Hydrophilic solutes are retained on that stagnant water-rich film depending on the difference of solvation compared to the mobile phase composition. However, the enhancement of selectivity by increasing the fraction of organic cosolvent is coupled with a limitation the analyte solubility, and the improvement of the HILIC principle by new hydrophilic stationary phases is the remaining option.
Y-zeolite (faujasite, FAU type) in the Na+-form with an average particle diameter of 5 μm was used as packing material in a 125 mm long HPLC column. The chromatographic response of the column was tested in methanol-water mixtures as eluent after injection of several aliphatic alcohols, polyols and monosaccharides with eluent conditions where no separation occurs on diol functionalized silica. On the zeolite the retention time increases according to ethylene glycol < glycerol < erythritol < sorbitol < inositol. The separation principle is explained to be superposed by two effects: firstly, a partition equilibrium between the water-rich phase in the zeolite micropores exists, and secondly, selective interactions with the inner crystalline pore surface and fixed-position Na+ ions, both serving to enhance the selectivity. Furthermore, arabinose and fructose monosaccharides could be separated into their tautomeric forms. Only upon increasing the temperature from 20 to 60 °C the tautomeric pattern merges into a single peak.
Instead of the stagnant water rich surface layer, zeolite micropores now take over that function. As a result, the selectivity among polyols and between α/β-arabinopyranose and β-fructopyranose/β-fructofuranose tautomers is extraordinary superior towards conventional hydrophilic interaction liquid chromatography (HILIC).
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•Zeolite as hydrophilic stationary phase can improve the HILIC principle.•Zeolite micropores serve as water-enriched volume instead of a water-enriched stagnant film in conventional HILIC stationary phases.•Separation of aliphatic alcohols and monosaccharide tautomers even with methanol as modifier where diol functionalized silica is ineffective.•Micropore dimension limits the separation principle to small molecules.
•A method to quantify ciprofloxacin in plasma by HPLC–UV.•Simple protein precipitation from serum with good recovery.•Validated method with acceptable reproducibility, precision, accuracy and ...stability.•Useful to quantify the concentration of ciprofloxacin in patients with Peripheral Arterial Disease.•Study will establish if an appropriate dosage regimen is being given to such patients.
A rapid and sensitive HPLC–UV method for the determination of ciprofloxacin in human plasma is described. Protein precipitation with acetonitrile was used to separate the drug from plasma protein. An ACE® 5 C18 column (250mm×4.6mm, 5μm) with an isocratic mobile phase consisting of phosphate buffer (pH 2.7) and acetonitrile (77:23, v/v) was used for separation. The UV detector was set at 277nm. The method was validated in the linear range of 0.05–8μg/ml with acceptable inter- and intra-assay precision, accuracy and stability. The method is simple and rapid and can be used to quantify this widely used antibiotic in the plasma of patients suffering from Peripheral Arterial Disease.
The role of polyphenolic compounds extractable from artichoke solid wastes in the formation of advanced glycation end products (AGEs) was studied. Outer bracts and stems were extracted using ...different water-ethanol mixtures and HPLC-DAD analyses indicated aqueous and hydro-alcoholic 20:80 stem extracts as the richest in polyphenols. The samples were characterized in their phenolic composition (using mass spectrometry) and antioxidant capacity. Antiglycative capacity was evaluated by in vitro BSA-sugars (glucose, fructose, and ribose) and BSA-methylglyoxal (MGO) tests, formation of Amadori products assay, direct glyoxal (GO) and MGO trapping capacity. Results indicated both extracts as effective inhibitors of fructosamine formation and antiglycative agents. In particular, aqueous extract showed the best activity in the systems containing glucose and fructose, differently from ethanolic extract, that was demonstrated able to better inhibit AGEs formation when ribose or MGO act as precursors. Ethanolic extract was also shown to be able to trap MGO and GO, with efficiency increasing after 24hours of incubation time. These activities are partially correlated with the antioxidant effect of the extract, as demonstrated by the scavenger capacity against ABTS cation and DPPH stable radicals; this relationship is evident when the model system, containing protein incubated with ribose or MGO, is considered. The different activities of the tested extracts could probably be ascribed to the different composition in chlorogenic acids (CQAs), being aqueous extract richer in 1-CQA, 3-CQA, and 1,3-di-CQA, and ethanolic extract in 5-CQA, caffeic acid, 1,5-di-CQA. These findings support further investigations to study the stability of the different CQAs in simil-physiological conditions and the feasibility of artichoke waste as antiglycative agents in food or pharmacological preparations.
5-caffeoylquinic acid (PubChem CID 5280633); 3-caffeoylquinic acid (PubChem CID 1794427); 1-caffeoylquinic acid (PubChem CID 10155076); 1,3-di-caffeoylquinic acid (PubChem CID 24720973); 1,5 – di-caffeoylquinic acid (PubChem CID 122685); caffeic acid (PubChem CID 689043); apigenin-7-glucuronide (PubChem CID 5319484); methylglyoxal PubChem CID (880); aminoguanidine hydrochloride (PubChem CID 2734687)
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•Chlorogenic acids from artichoke wastes inhibited protein glycation.•Artichoke waste polyphenols directly trapped methylglyoxal.•Mono/di-caffeoylquinic acids from artichoke wastes inhibited fructosamine formation.•Re-cycling of artichoke by-products is useful in the production of nutraceuticals.
