High-performance Liquid Chromatography Studies Research Articles

B-179 Comparative Evaluation of Cisplatin Release from Alginate Hydrogels with Embedded Silver Nanoparticles: An HPLC and Colorimetric Spectrophotometry Study

Abstract Background Cisplatin [cis-dichlorodiamine platinum (II)], is a well-recognized chemotherapeutical drug. Cisplatin has been employed in treating a wide range of human cancers, such as those of the breast, bladder, lung, ovarian, and testicular cancers. Its therapeutic action is attributed to its capacity to form crosslinks with the DNA's purine bases, disrupting DNA repair processes, causing DNA damage, and ultimately leading to apoptosis in cancerous cells. Nonetheless, the application of cisplatin is constrained by the development of multidrug resistance and the occurrence of severe adverse effects, including nephrotoxicity, bone marrow suppression, hearing loss, allergic responses, nerve damage, and low magnesium levels in the blood. Numerous strategies have been explored to enhance the anticancer effectiveness of cisplatin while minimizing its associated toxicities. These strategies include combination therapies that incorporate nanoparticles, liposomes, and polymer micelles. In this project, cisplatin was embedded into alginate hydrogels loaded with silver nanoparticles and in vitro cisplatin release study using HPLC and UV-Vis spectrophotometry were studied. Methods We developed a novel nanocomplex by integrating silver nanoparticles (AgNPs) with alginate hydrogel coating to create a versatile platform for drug delivery. To incorporate the chemotherapeutic agent, cisplatin (150 ppm) was introduced into the AgNPs-alginate mixture utilizing rapid stirring to ensure uniform distribution and encapsulation of cisplatin within the nanocomplex. A range of analytical methods, such as UV-Vis, FTIR, SEM/EDX, and Zeta potential analysis, were employed to characterize the nanocomplex. To evaluate cisplatin release kinetics from the nanocomplex, we employed an in vitro dialysis method for monitoring its sustained release. High-performance liquid chromatography (HPLC) was used for precise cisplatin release quantification, then validated by colorimetric spectrophotometry. This dual-method approach ensured accurate insights into the nanocomplex’s release dynamics, substantiating its potential in enhancing targeted cancer therapy through advanced drug delivery systems. Results The analysis through UV-Vis revealed an absorption spectrum around 410 nm, indicative of the characteristic plasmon resonance associated with silver nanoparticles. TEM provided high-resolution imagery, revealing that the size of the silver nanoparticles varied between 4 and 30 nm, averaging at 13 nm with a standard deviation of 5 nm specifically for the alginate-coated AgNPs. SEM images confirmed the anticipated spherical distribution of silver nanoparticles within the alginate hydrogel framework. EDX spectroscopy analysis further verified the incorporation of silver nanoparticles and platinum within the complex. The cisplatin release studies from this newly developed nanocomplex illustrated a prolonged, slow, and consistent release pattern, extending over days, in stark contrast to the rapid and complete release of cisplatin from its unbound state, which achieved peak release within an hour. Conclusions In this study, we successfully developed and validated an innovative, multifunctional nanoplatform for drug delivery, comprising alginate hydrogel synergistically co-loaded with silver nanoparticles (AgNPs) and cisplatin. This novel drug carrier system demonstrated a marked enhancement in the controlled delivery of cisplatin, evidenced by the slow and sustained release profile observed over three days. The slow and sustained release of cisplatin from the alginate complex, compared to the free form, highlights the efficacy of the nanoplatform in modulating the drug’s release kinetics.

HPLC and LC-MS/MS-Based Quantitative Characterization of Related Substances Associated with Sotalol Hydrochloride.

In total, three related substances (RS) associated with sotalol hydrochloride (STHCl) were herein identified with a novel gradient high-performance liquid chromatography (HPLC) protocol. Further characterization of these substances was then performed via liquid chromatography-mass spectroscopy (LC-MS/MS) and nuclear magnetic resonance (NMR) approaches. For these analyses, commercial STHCl samples were used for quantitative HPLC studies and the degradation of STHCl under acidic (1M HCl), alkaline (1M NaOH), oxidative (30% H2O2), photolytic (4500 Lx), and thermal stress conditions (100 °C) was assessed. This approach revealed this drug to be resistant to acidic, alkaline, and high-temperature conditions, whereas it was susceptible to light and oxidation as confirmed through long-term experiments. The putative mechanisms governing RS formation were also explored, revealing that RS3 was derived from the manufacturing process, whereas RS2 was generated via oxidation and RS1 was generated in response to light exposure. The cytotoxicity of these RS compounds was then assessed using MTT assays and acute toxicity test. Overall, this study provides details regarding the characterization, isolation, quantification, and toxicological evaluation of STHCl and associated RS compounds together with details regarding the precise, specific, and reliable novel HPLC technique, thus providing the requisite information necessary to ensure STHCl purity and safety.

Compatible Kinetic Model for Quantitative Description of Dual-Clock Behavior of the Complex Thiourea-Iodate Reaction.

The thiourea-iodate reaction has been investigated simultaneously by ultraviolet-visible spectroscopy and high-performance liquid chromatography (HPLC). Absorbance-time traces measured at the isosbestic point of the iodine-triiodide system have revealed a special dual-clock behavior. During the first kinetic stage of the title reaction, iodine suddenly appears only after a well-defined time lag when thiourea is totally consumed due to the rapid thiourea-iodine system giving rise to a substrate-depletive clock reaction. After this delay, iodine in the system starts to build up suddenly to a certain level, where the system remains for quite a while. During this period, hydrolysis of formamidine disulfide as well as the formamidine disulfide-iodine system along with the Dushman reaction and subsequent reactions of the intermediates governs the parallel formation and disappearance of iodine, resulting in a fairly constant absorbance. The kinetic phase mentioned above is then followed by a more slowly increasing sigmoidally shaped profile that is characteristic of autocatalysis-driven clock reactions. HPLC studies have clearly shown that the thiourea dioxide-iodate system is responsible mainly for the latter characteristics. Of course, depending on the initial concentration ratio of the reactants, the absorbance-time curve may level off or reach a maximum followed by a declining phase. With an excess of thiourea, iodine may completely disappear from the solution as a result of the thiourea dioxide-iodine reaction. In the opposite case, with an excess of iodate, the final absorbance reaches a finite value, and at the same time, iodide ion will disappear completely from the solution due to the well-known Dushman (iodide-iodate) reaction. In addition, we have also shown that in the case of the formamidine disulfide-iodine reaction, unexpectedly the triiodide ion is more reactive toward formamidine disulfide than iodine. This feature can readily be interpreted by the enhancement of the rate of formation of the transition complex containing oppositely charged reactants. A 25-step kinetic model is proposed with just 10 fitted parameters to fit the 68 kinetic traces measured in the thiourea-iodate system and the second, but slower, kinetic phase of the thiourea-iodine reaction. The comprehensive kinetic model is constituted in such a way as to remain coherent in quantitatively describing all of the most important characteristics of the formamidine disulfide-iodine, thiourea dioxide-iodine, and thiourea dioxide-iodate systems.

Identification of secondary metabolites and fatty acids in Curculigo orchioides Gaertn., an important ethnomedicinal plant

Curculigo orchioides Gaertn, is a herb belonging to family Amaryllidaceae. The plant possesses aromatic smell and used by ethnic community (Majhi, Munda, Santal, Birhor, Ho and Rajwar) from Purulia district, West Bengal, India as a whole either topically or orally in various monoherbal and polyherbal formulations for the treatment of various skin diseases such as carbuncle, acne, pimple and ulcer. However, there is no scientific database on secondary metabolites and fatty acids of the plant yet. So, main emphasis of the present study is given on secondary metabolites and fatty acids of the plant. Secondary metabolites and fatty acids were analyses by HPLC (High-performance Liquid Chromatography) and GCMS (Gas Chromatography Mass Spectrometry). HPLC study revealed the presence of phenolic compounds and flavonoids. GCMS analyses identified the presence of saturated fatty acids in the range of C14 to C26 and C18 and C20 as unsaturated fatty acids. Assured levels of secondary metabolites along with fatty acids in the studied plant part support the ethnobotanical claim done by traditional healers.

Antihyperlipidemic activity of hydroalcoholic seed extract of Cucumis sativus L in triton X-100 induced hyperlipidemic rats

Hyperlipidemia is linked to an elevated risk of atherosclerosis disease and managing this risk factor is crucial. This study was aimed at investigation of high performance liquid chromatographic (HPLC) analysis and antihyperlipidemic effect of hydroalcoholic extract of seeds of the plant Cucumis sativus L in Wistar albino rats. The rats were divided into groups A, B, C and D. All the groups except group A were administered with diet having high cholesterol and given intraperitoneal injection of Triton X-100 to induce hyperlipidaemia. Group A served as normal. Group B served as hyperlipidemic control. Group C (standard) and D (plant treated) were administered with simvastatin (10 mg/kg) and hydroalcoholic extract of the plant (400 mg/kg) respectively daily for 7 days after one week of inducing hyperlipidemia. The biochemical parameters of all the groups were estimated after one week of administration of plant extract using kits of Roche Cobas C111 chemistry analyzer. Histological studies of liver and heart tissues were also done. HPLC study showed the presence of compounds in the extract. The plant decreased the levels of total cholesterol, triglycerides, alanine aminotransferase, low and very low-density lipoprotein cholesterol significantly. The plant may be used as dietary supplement in combating hyperlipidemia.

Open-Label Study to Evaluate the Efficacy of a Topical Anhydrous Formulation with 15% Pure Ascorbic Acid and Ginger as a Potent Antioxidant

Vitamin C is one of the naturally occurring antioxidants capable of reducing or preventing skin photoaging. Achieving a stable formulation with the optimal dose of ascorbic acid to ensure a biologically significant antioxidant effect is a challenge when developing cosmetic formulations. The objective of this study was to develop a stable formula in a non-aqueous media with 15% pure vitamin C supplemented with ginger and to study its efficacy, skin tolerance, and cosmetic assessment in 33 women. Vitamin C stability over time was determined via a high-performance liquid chromatography (HPLC) technique versus an aqueous option. Reactive oxygen species (ROS) determination was quantified to provide antioxidant effect. A 56-day in vivo study was performed to evaluate skin luminosity and hyperpigmentation reduction. Skin acceptability was verified by a dermatologist. The HPLC studies demonstrated a high stability of the anhydrous formula compared to an aqueous option. The in vitro studies showed a reduction in ROS of 93% (p-value < 0.0001). In vivo, luminosity increased by 17% (p-value < 0.0001) and skin tone became 10% more uniform (p-value < 0.007). Moreover, very good skin tolerance was determined as the dermatologist did not determine any clinical signs, and the subjects did not report any feelings of discomfort. We were able to develop an anhydrous formula of pure vitamin C that combines very good stability, consumer acceptance, and skin tolerance with a high level of efficacy.

Petasites japonicus extract exerts anti-malarial effects by inhibiting platelet activation

BackgroundNew antimalarial agents are needed to combat emerging resistance to the currently available drugs. In the pathology of cerebral malaria, platelets play a central role by binding infected and uninfected red cells and the endothelium. Since Petasites japonicus extract was reported as an effective inhibitor of platelet activation, we examined the antimalarial activities of the P. japonicus extract. PurposeThis study aimed to evaluate the impact of P. japonicus extract prepared from whole plants on malarial infection. MethodsThe P. japonicus extract were characterized by high‐performance liquid chromatography (HPLC) profiling. Antimalarial activity of the P. japonicus ethanolic extract was evaluated in vitro using chloroquine-sensitive (3D7) and chloroquine-resistant (Dd2) P. berghei strains. Also, the in vivo activity of the extract was evaluated in P. berghei-infected mice via oral administration followed by a four-day suppressive test to measure the hematological parameters. In addition, platelet activation signaling induced by the P. japonicus extract in P. berghei infection was evaluated. Results: In HPLC study, catechin, rutin, liquiritin, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid were identified in P. japonicus extract. Exposure to the P. japonicus extract significantly inhibited both CQ-sensitive (3D7) and resistant (Dd2) strains of P. falciparum with IC50 values of 8.48 ± 1.70 and 7.83 ± 6.44 μg/ml, respectively. Administration of the P. japonicus extract also resulted in potent antimalarial activities in P. berghei-infected mice with no associated toxicity. The treatment also improved the hematologic parameters. In addition, the survived mice from P. berghei infection exhibited the inhibition of collagen-induced platelet aggregation by attenuated glycoprotein VI (GPVI) downstream signaling. Conclusion:P. japonicus extracts promote antimalarial effects both in vitro and in vivo. In addition, the effects appear to be induced by the inhibition of collagen-induced platelet activation related to attenuated GPVI downstream signaling. Further studies to identify and characterize the antimalarial compounds in P. japonicus will promote the development of new drugs.

Cow and Ewe Cheeses Made with Saffron: Characterization of Bioactive Compounds and Their Antiproliferative Effect in Cervical Adenocarcinoma (HeLa) and Breast Cancer (MDA-MB-231) Cells.

Saffron is a widespread consumed spice containing many phytochemicals. It is often used in dairy technologies to enhance color and flavor of cheeses, but it is also known for its several therapeutic effects, as well as its antiproliferative and anticancer properties. In this study High Performance Liquid Chromatography was used to characterize saffron bioactive compounds in cow and ewe cheeses made with saffron, and the antiproliferative effect of the crocin-rich extracts from cheeses was investigated on different cellular lines (CaCo2, MDA-MB-231 and HeLa) by MTT assay. Crocins were observed in all cheese samples, with the total content ranging between 0.54 and 30.57 mg trans-4-GG/100 g cheese, according to the different cheese making process. Picrocrocin was detected in no cheese (probably due to its degradation during cheese making), while safranal was detected only in one ewe cheese (mainly due to its high volatility). HeLa and MDA-MB-231 cells were sensitive to treatment with crocin-rich extracts from cheeses, while no effect was observed on CaCo2 cells. The chemical environment of the food matrix seems to have a great influence on the crocin antiproliferative effect: the crocin-rich extracts from cheese with both high residual N/protein and fat contents showed increased antiproliferative effect compared to pure crocin (trans-4-GG), but cheeses from different milk species (type of fats and proteins) could also play an important role in modulating crocin’s antiproliferative effects.

The hematological and histological studies for the hepatoprotective-like effect of the hydromethanolic extract and the fractions of Viola serpens Wall.

Traditionally, Viola serpens has been used in the treatment of several human disorders including liver diseases without any scientific evidence. As the current therapies are not very effective and face challenges of unwanted effects and patient compliance, therefore more effective and safe agents are highly needed. The current study aimed to evaluate the hepatoprotective potential of the crude extract and subsequent fractions of the whole plant in the in-vivo model using various hematological and histopathological parameters followed by an HPLC study for the identification of phenolic compounds. Rabbits (1000-1200 g) were used in the study. Paracetamol (2g) was used to induce hepatotoxicity in experimental rabbits. The plant extract was used in two doses (150 and 300 mg/kg body weights) for eight days. The hematological parameters AST, ALT and ALP values were determined along with the histopathology of the liver. Phenolic compounds were identified by high-performance liquid chromatography (HPLC) Agilent-1260 infinity from their retention time, UV spectra and available standards while quantification was done taking the percent peak area. The doses 150 and 300 mg/kg body weight seemed to be more effective. The hematological values and the histopathological slides show the hepatoprotective effect of the plant. Regeneration indicated the presence of nuclei, nuclear cleaning, prominent nucleoli, RBC's, central veins and plates of hepatocytes. The HPLC studies revealed the presence of a number of phenicol compounds. The crude extract and the subsequent fractions of the plant possess strong hepatoprotective activity, providing a scientific rationale for its uses in the treatment of liver toxicities.

HPLC profiling and studies on Copaifera salikounda methanol leaf extract on phenylhydrazine-induced hematotoxicity and oxidative stress in rats

Anemia is a clinical disorder orchestrated by factors such as drugs, including phenylhydrazine (PHZ). This study explored the protective roles of Copaifera salikounda methanol leaf extract (CSMLE) against PHZ-induced hematotoxicity and oxidative stress in rats. In vitro antioxidants studies of CSMLE using 1,1-diphenyl-2-picryl hydrazyl (DPPH) and ferric reducing antioxidant potential (FRAP) and quantification of CSMLE bioactive compounds using High-Performance-Liquid-Chromatography (HPLC) were done. In vivo , thirty rats in five groups (n = 6) were used. Anemia was induced intraperitoneally in groups 2 to 5 with 40 mg/kg body-weight/day PHZ for two days. Group 1 received normal saline, group 2 (untreated: negative-control), group 3 received 1 ml/kg-body-weight Vit B 12 syrup while groups 4 and 5 received 200 mg/kg and 400 mg/kg-body-weight CSMLE, respectively, daily for fourteen days. Result of CSMLE profiling showed luteolin (76.49 ng/Ml) and squalene (38.31 ng/Ml) as the most abundant flavonoid and terpenoid, respectively. In vitro CSMLE demonstrated dose-dependent antioxidant effects on DPPH and FRAP with IC 50 of 0.1085 mg/ml and 0.07344 mg/ml (EC 50 ), respectively. In vivo, CSMLE improved the body weight, red blood cell, hematocrit and hemoglobin concentrations, significantly (P < 0.05), reduced serum malondialdehyde and nitic oxide but increased reduced glutathione levels, superoxide dismutase and catalase activities and attenuated the levels of Interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in dose-dependent manner in treated groups relative to the negative control. The spleen tissues histology results corroborated biochemical results. The results indicated that CSMLE possesses anti-anemic potentials and attenuate spleenotoxic effects of PHZ.

Cassia auriculata and its role in infection / inflammation: A close look on future drug discovery

The herbal plants contain various active compounds which have the ability to treat several human diseases without any side effects like cancer chemotherapeutic drugs used currently. It is reported that majority of the presently used drugs are derived from natural resources like plants or their products and hence, there is an urgent need to search for novel drugs from natural resources. Accordingly, the present study was designed to exploit an ethnomedicinally important plant Cassia auriculata Linn. for the identification of its phytoconstituents responsible for various properties such as antioxidant, anti-microbial and anti-diabetic activities. Different extracts of the plant were subjected to chromatographic techniques like high-performance liquid chromatography (HPLC), gas chromatography, and mass spectrometry (GC-MS) analysis to acquire the fingerprint of the phytoconstituents present in the plant. The extracts were analyzed by Ultra Violet-Visible (UV–Visible) and Fourier-Transform Infrared Spectroscopy (FT-IR). The efficacy of the extracts was examined through different in vitro assay methods. The GC-MS study exhibited the occurrence of 48 phytoconstituents in all the tested extracts and the HPLC study revealed the presence of quercetin. Different in vitro evaluations of the plant revealed that both ethyl acetate (DPPH-IC50: 340.9 μg/ml) and ethanol fractions (DPPH-IC50: 205.5 μg/ml) exhibited a potent activity. Hence, using the above study, novel potent antimicrobial and anti-diabetic principles from C. auriculata can be formulated in the future towards the clinical progress of therapeutic remedies against these ailments.

Determination of the Physicochemical Properties of Piroxicam.

The aim of this study was to determine the acid dissociation constant (pKa) of piroxicam using high performance liquid chromatography (HPLC) and ultraviolet-visible (UV-Vis) spectrophotometry, and to determine the partition coefficient (Log P), distribution coefficient (Log D), and "Log kw" values of piroxicam using HPLC. The HPLC studies were performed on a reversed-phase ACE C18 (150x4.6 mm ID, 5 μm) column at a flow rate of 1.0 mL min-1. The detector was set to 360 nm. Log D at different pH values (3.0-6.5) was examined with a phosphate buffer (20 mM) and acetonitrile (30:70 v/v) mixture as the mobile phase. For pKa determination, HPLC studies were performed with a mixture of phosphate buffer (20 mM) and methanol within the pH range of 3.50-6.00. Log kw measurements were performed with phosphate buffer (20 mM) and MeOH (from 20:80 v/v to 10:90 v/v) mixtures within the pH range of 3.50-6.00. UV-Vis spectrophotometric pKa measurements were performed at 285 nm wavelength. The pKa value of piroxicam was found to be 5.3 by HPLC and 5.7 by UV-Vis spectrophotometry. Log P of piroxicam was determined as 1.58 in our experimental conditions. Log D values were 1.57, 1.57, 1.44, 1.13, and 0.46 for pH values of 3.17, 3.79, 4.44, 5.42, and 6.56, respectively. In the literature, different Log P (3.1, 2.2, and 0.6) and pKa (6.3 and 4.8) values were reported for piroxicam. The Log P (1.58) and pKa (5.3 and 5.7) values obtained for piroxicam in our study were within the range of the literature values. All these results indicate that different experimental approaches used for the determination of physicochemical properties could provide different values. Although UV spectrophotometry is easy to apply, HPLC is a unique technique for simultaneous determination of pKa, Log D, and Log P values of compounds.

Standardization of Optimized Anti-diabetic Polyherbal Formulation by HPLC

Herbal drugs are widely used to treat and prevent various diseases with more than 80% of people worldwide relying on them for their primary healthcare. Polyherbal formulations express high effectiveness in various diseases due to their synergistic effect. Herbal formulations are required to standardize in order to assess their quality, purity, and biological efficacy. The present study aimed to prepare a polyherbal formulation comprising of lyophilized ethanolic extracts of three well known anti-diabetic herbal drugs (Momordica charantia, Andrographis paniculata and Withania somnifera), and three formulations were developed in combination ratios (1:1:1, 2:1:2, and 2:2:1) respectively and to develop high-performance liquid chromatography (HPLC) method for assessment of purity, standardization and estimation of markers. In addition, developed formulations were optimized on the basis of oral glucose tolerance test (OGTT). Standardization of developed polyherbal formulations was performed by using respective biological markers (Vicine, andrographolide, withaferin-A and withanolide-A) by HPLC technique. HPLC was performed using RP-18, Merck column to study the presence and quantification of respective markers at a wavelength of 237 nm. The HPLC study revealed the presence of vicine, andrographolide, withaferin-A, and withanolide-A in quantifiable amounts in respective extracts. OGTT study was performed using normal albino Wistar rats at a drug dose of 500 mg/kg body weight. Rats were divided into six groups, including three combination ratios and standard (Metformin) group. An optimized ratio 2:2:1 showed a significant decrease in blood glucose levels compared with standard control rats.

Broad-Spectrum Cytotoxic Effect of Calendula officinalis L Against Breast Cancer Cells

Background: Calendula officinalis L used in Iraqi folklore medicine for several medical applications. This research evaluated the leaves extract as an anti-breast cancer agent in in-vitro cancer cell line systems and studies its active compounds. Crystal violet viability assay was used to determine the cytotoxicity of the leave methanolic extract of Calendula officinalis L against diverse breast cancer cell lines. Human breast cancer MCF7, AMJ13, MDAMB, and CAL51 cells were treated with different concentrations of extract for 72 hours. Morphological study for the exposed cell was done by examination under a phase-contrast inverted microscope. High-performance liquid chromatography (HPLC) analysis was performed to measure the concentrations of each component of phenols and flavonoids in the Calendula officinalis L extract. Results: It was found that methanolic extract of Calendula officinalis L inhibits the proliferation of all breast cancer cells significantly at the meantime; it does not affect normal embryonic cells. Additionally, it induced the cytopathic morphological changes in cancer cells. Furthermore, HPLC study revealed that Calendula officinalis L extract contained an important component of flavonoids. Conclusions: Calendula officinalis L leaves extract inhibited the proliferation of breast cancer cells especially MDAMB cells with no effect on normal cells. This work showed that Calendula officinalis L is a possible natural source as broad-spectrum anti-breast cancer drug.

Broad-Spectrum Cytotoxic Effect of Calendula officinalis L Against Breast Cancer Cells

Background: Calendula officinalis L used in Iraqi folklore medicine for several medical applications. This research evaluated the leaves extract as an anti-breast cancer agent in in-vitro cancer cell line systems and studies its active compounds. Crystal violet viability assay was used to determine the cytotoxicity of the leave methanolic extract of Calendula officinalis L against diverse breast cancer cell lines. Human breast cancer MCF7, AMJ13, MDAMB, and CAL51 cells were treated with different concentrations of extract for 72 hours. Morphological study for the exposed cell was done by examination under a phase-contrast inverted microscope. High-performance liquid chromatography (HPLC) analysis was performed to measure the concentrations of each component of phenols and flavonoids in the Calendula officinalis L extract. Results: It was found that methanolic extract of Calendula officinalis L inhibits the proliferation of all breast cancer cells significantly at the meantime; it does not affect normal embryonic cells. Additionally, it induced the cytopathic morphological changes in cancer cells. Furthermore, HPLC study revealed that Calendula officinalis L extract contained an important component of flavonoids. Conclusions: Calendula officinalis L leaves extract inhibited the proliferation of breast cancer cells especially MDAMB cells with no effect on normal cells. This work showed that Calendula officinalis L is a possible natural source as broad-spectrum anti-breast cancer drug.

The Aquilegia pubiflora (Himalayan columbine) mediated synthesis of nanoceria for diverse biomedical applications.

Herein, we report an eco-friendly, facile, one-pot, green synthesis of nanoceria for multiple biomedical applications. In the study, cerium oxide nanoparticles (CeO2-NPs) were synthesized using a simple aqueous extract of Aquilegia pubiflora as an effective reducing and capping agent. The biosynthesized nanoparticles were characterized via UV-vis spectroscopy, X-ray powder diffraction (XRD), high-performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and Raman spectroscopy. The NPs were highly stable, exhibited high purity, and had a spherical morphology and mean size of 28 nm. FTIR and HPLC studies confirmed the successful capping of bioactive compounds on the nanoparticles. The well-characterized NPs were evaluated for a number of biomedical applications, and their antimicrobial (antifungal, antibacterial, and antileishmanial), protein kinase inhibition, anticancer, antioxidant, anti-diabetic and biocompatibility properties were studied. Our results showed that the Aquilegia pubiflora mediated CeO2-NPs were highly active against fungal strains, compared to the tested bacterial strains, with Aspergillus niger resulting in the largest zone of inhibition (15.1 ± 0.27 mm). The particles also exhibited dose dependent leishmanicidal activity with significant LC50 values toward both the amastigote (114 μg mL−1) and promastigote (97 μg mL−1) forms of the parasite Leishmania tropica (KWH23). The NPs were found to be moderately active against the HepG2 cell line, showing 26.78% ± 1.16% inhibition at 200 μg mL−1. Last but not least, their highly biocompatible nature was observed with respect to freshly isolated human red blood cells (hRBCs), making the greenly synthesized CeO2-NPs a novel candidates for multidimensional medical applications.

Selective cytotoxic effect of Plantago lanceolata L. against breast cancer cells

BackgroundPlantago lanceolata L. is used in Iraqi folklore medicine to treat injuries, and its extract is prescribed by some herbalists for cancer patients. This research aimed to evaluate the effect of P. lanceolata leaf extract on breast cancer cell lines in vitro and to identify its active compounds. Crystal violet viability assay was used to determine the cytotoxicity of methanolic P. lanceolata leaf extract against various breast cancer cell lines. MCF7, AMJ13, MDAMB, and CAL51 human breast cancer cells were treated with different concentrations of the extract for 72 h. The morphology of the treated cells was examined under a phase-contrast inverted microscope. The clonogenic ability was assessed through a clonogenic assay. High-performance liquid chromatography (HPLC) analysis was performed to measure the concentrations of phenols and flavonoids in the extract.ResultsThe methanolic P. lanceolata leaf extract significantly inhibited the proliferation of triple-negative CAL51 cells but showed minor effect on the other breast cancer cells. In addition, at high doses, it induced cytopathic morphological changes. The clonogenic assay showed low colony formation in the exposed cells, especially CAL51 cells. Furthermore, HPLC study revealed that the methanolic extract contained important flavonoid glycosides, especially rutin, myricetin quercetin, and kaempferol.ConclusionsP. lanceolata leaf extract selectively inhibited the proliferation of CAL51 triple-negative breast cancer cells and showed minor effect on the other breast cancer cells types studied. Thus, this study showed P. lanceolata as a possible natural source of selective anti-triple-negative breast cancer drugs.

Electrochemical behavior of esomeprazole: Its determination at Au electrode as standard and in injection powder combined with the study of its degradation

Esomeprazole is the most effective of the proton-pump inhibitors for the acid-related diseases and at first was examined for the electroanalytical purposes. The drug standard and as a content of injection powder was investigated by cyclic voltammetry (CV) and quantitatively determined using square wave voltammetry (SWV) via its electrooxidation at Au electrode in 0.05M NaHCO3. SWV showed a linear dependency of the anodic peak currents vs. esomeprazole standard concentrations in the range from 3.0 to 500μgmL−1 with the values of limit of detection (LOD) and limit of quantification (LOQ): 1.4 and 4.6μgmL−1, respectively. Using the constructed and validated calibration curve, the values of unknown esomeprazole concentrations in injection powder and in human serum spiked with standard were determined. Before the electrochemical oxidation, it was shown by atomic force microscopy (AFM) that the small esomeprazole islands formed inside holes were visible and their diameter was about 200nm attributed to physico-chemical characteristics of esomeprazole. After the electrochemical oxidation, the morphology of esomeprazole standard on Au surface was completely changed and composed of spherical particles in a diameter between 200 and 600nm. With esomeprazole suspended in human serum, the process of crystallization partly occurred in the form of spherical grains with the average size of these grains was about 4μm. The analysis at the macro level done by the optical microscopy (OM) confirmed this opinion.The study of esomeprazole degradation showed that at Au electrode, after 3h of cycling, a neglectable amount of the esomeprazole was changed. Using IrOx electrode under directed stress conditions, its almost complete degradation was realized after 3h confirmed by high performance liquid chromatography (HPLC). Total organic carbon (TOC) analysis showed that 95% of esomeprazole was mineralized. The HPLC and Liquid chromatography-mass spectrometry (LC-MS) study revealed the formation of 4-hydroxy omeprazole sulphide, 4-hydroxy omeprazole sulphone, esomeprazole sulphone and methylated esomeprazole.

Comparative high-performance liquid chromatographic and high-performance thin-layer chromatographic study for the simultaneous determination of dapagliflozin and metformin hydrochloride in bulk and pharmaceutical formulation

The discovery of potent antidiabetic drugs is of necessity owing to the rapid prevalence of diabetes worldwide. The investigation of a new separation method for the simultaneous determination of combined antidiabetic drugs is thus an essential issue to cover the usual demands of simple analytical methods for the routine analysis. Therefore, herein, simple and fast chromatographic methods were established for synchronized determination of metformin (MET) and dapagliflozin (DAP), a mixture approved recently by the Food and Drug Administration (FDA) for diabetes therapy. In high-performance liquid chromatography (HPLC), a Water-pak C18 column was used as the stationary phase with an isocratic mobile phase composed of 10 mM NaH2PO4 (pH 3.5 adjusted using ortho-phosphoric acid)‒acetonitrile (65:35, v/v) containing 0.1% trimethylamine at a flow rate of 1.2 mL min−1 and a wavelength detector set at 225 nm. In high-performance thin-layer chromatography (HPTLC), separation was achieved on pre-coated silica gel 60 ...

Glutamate, Glutamine and GABA Levels in Rat Brain Measured Using MRS, HPLC and NMR Methods in Study of Two Models of Autism.

The disorders of the glutamatergic neurotransmission have been associated with pathogenesis of autism. In this study we evaluated the impact of the in vivo and ex vivo test methodology on measurements of levels of neurotransmitter amino acids in hippocampus of rats for valproic acid- (VPA) and thalidomide- (THAL) induced models of autism. The main goal was to compare the changes in concentrations of glutamate (Glu), glutamine (Gln) and GABA between both autistic groups and the control, measured in vivo and ex vivo in homogenates. The rat pups underwent three in vivo tests: ultrasonic vocalization (USV), magnetic resonance spectroscopy (MRS) and unilateral microdialysis of the hippocampus. Analyses of homogenates of rat hippocampus were performed using high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) spectroscopy. For the statistical analysis, we performed univariate and multivariate tests. USV test, which is considered in rodents as an indicator of pathology similar to autism, showed decreased USV in VPA and THAL groups. In vivo MRS studies demonstrated increases of Glu content in male rat’s hippocampus in VPA and THAL groups, while the microdialysis, which allows examination of the contents in the extracellular space, detected decreases in the basal level of Gln concentrations in VPA and THAL groups. Ex vivo HPLC studies showed that levels of Glu, Gln and GABA significantly increased in male rat’s hippocampus in the VPA and THAL groups, while NMR studies showed increased levels of Gln and GABA in the VPA group. Collectively, these results are consistent with the hypothesis suggesting the role of the glutamatergic disturbances on the pathogenesis of autism. For all methods used, the values of measured changes were in the same direction. The orthogonal partial least square discriminant analysis confirmed that both animal models of autism tested here can be used to trace neurochemical changes in the brain.