HPLC Procedure Research Articles

Nanofabrication of PLGA-PEG-chitosan-folic acid systems for delivery of colchicine to HT-29 cancer cells

This survey was conducted to fabrication of PLGA-based nanosystems modified with PEG, chitosan and folic acid to delivery colchicine to cancer cells and to investigate its antioxidant and pro-apoptotic effects. The dual emulsion-evaporation solvent method was used for loading of colchicine on PEGylated PLGA nanoparticles (COL-PP-NPs) and after surface modification with chitosan and folic acid (COL-PPCF-NPs), the nanoparticles were characterized by DLS, SEM and FTIR methods. The HPLC procedure was used to assess the amount of FA binding and COL loading. Antioxidant capacity (ABTS and DPPH free radical scavenging) and toxicity (MTT) of COL-PPCF-NPs were evaluated and then cell inhibition mechanism was assessed by AO/PI staining, flow cytometry and qPCR assay. COL-PPCF-NPs with a size of 250 nm were synthesized in a stable (zeta potential: +34 mV) and mono-dispersed (PDI: 0.32) manner. FA binding and COL loading were reported to be 55% and 89.5%, respectively. COL-PPCF-NPs showed antioxidant effects by inhibiting the free radicals ABTS (108.07 µg/ml) and DPPH (361.61 µg/ml). The selective toxicity of COL-PPCF-NPs against HT-29 cancer cells (118.5 µg/ml) compared to HFF cells was confirmed by MTT data. Increased apoptotic cells (red color) in AO/PI staining, cell arrest in phase SubG1 and G2-M, and altered expression of apoptosis genes confirmed the occurrence of apoptosis in HT-29 treated cells. The use of PPCF-NPs system for delivery of COL can lead to selective toxicity against cancer cells and induction of apoptosis in these cells by folate-mediated binding mechanism at folate receptor positive HT-29 cancer cells.

Analytical Method Development and Validation Analysis for Quantitative Assessment of Thifluzamide by HPLC Procedure

Analytical Method Development and Validation Analysis for Quantitative Assessment of Thifluzamide by HPLC Procedure

Multi-analyte HPLC–DAD Method for Concurrent Analysis of Six Antimicrobials and Three Proton Pump Inhibitors Frequently used in Management of Helicobacter pylori Infection: Application to Simulated Intestinal Fluid Samples

The present work deals with the optimization, validation and application of a versatile HPLC–DAD method for concurrent estimation of nine antimicrobials and proton pump inhibitors, namely amoxicillin (AMX), doxycycline (DOX), furazolidone (FRZ), lansoprazole (LNS), levofloxacin (LVF), metronidazole (MTZ), omeprazole (OMZ), pantoprazole (PNZ) and tinidazole (TNZ). The selected nine drugs are frequently included in various treatment regimens of Helicobacter pylori infection. Successful separation was accomplished using the analytical column Agilent Zorbax Eclipse plus-C18 (250 × 4.6 mm, 5 µm particle size) and a mobile phase prepared from phosphate buffer pH 5 and acetonitrile pumped at a flow rate 1 mL/min using a gradient elution program. The gradient elution started with buffer/acetonitrile ratio 90:10, then it was altered in 15 min to reach 40:60 by volume. Quantification of the analytes was based on measuring peak areas of AMX at 230 nm, LVF, LNS and PNZ at 290 nm, OMZ at 300 nm, MTZ and TNZ at 320 nm, and DOX and FRZ at 360 nm. The separated compounds eluted at retention times 5.68, 6.43, 7.82, 8.84, 9.42, 10.75, 12.82, 13.74 and 14.90 min for AMX, MTZ, LVF, TNZ, DOX, FRZ, OMZ, PNZ and LNS respectively. Validation of the proposed HPLC procedure was carefully studied according to the ICH items: ranges, precision, accuracy, linearity, robustness and limits of detection and quantitation. The linear dynamic ranges were 5–100, 5–50, 2–40, 10–100, 10–100, 5–50, 2.5–30, 3–30 and 2–30 µg/mL for AMX, MTZ, LVF, TNZ, DOX, FRZ, OMZ, PNZ and LNS, respectively with correlation coefficients > 0.9993. Application fields of the validated method included analysis of laboratory-prepared binary dosage forms along with analysis of several ternary mixtures in spiked simulated intestinal fluid.

Development of a simple HPLC procedure for the determination of prochloraz residues in mushrooms

Abstract A simple HPLC-UV procedure is described in our paper which is suitable for the rapid and cost-efficient determination of prochloraz in mushrooms. Prochloraz is the only fungicide in EU which use is allowed in mushroom production. The aim of our work was the development of a simple method that is suitable for the control of this pesticide in everyday analyses during mushroom production. The procedure involves a simple sample preparation method based on solid-liquid extraction (modified QuEChERS extraction method EN 15662) followed by an HPLC-UV determination (recovery: 97–99%; limit of detection LOD: 0.01 mg/kg; limit of quantification LOQ: 0.05 mg/kg).

Dispersive micro solid phase extraction based ionic liquid functionalized ZnO nanoflowers couple with chromatographic methods for rapid determination of aflatoxins in wheat and peanut samples

Aflatoxins (AFs) contaminate agricultural products in a wide range of ways during their harvesting, storage and transport. Therefore, the detection of AFs has certain practical significance. Herein, a dispersive micro solid phase extraction (D-µSPE) technology was constructed based on 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIM][PF6]) fabricated ZnO nanoflowers for AFs extraction from food matrix before HPLC procedure. The key parameters affecting the extraction efficiency were studied. Under optimal experimental conditions, the method showed excellent linearity with high correlation coefficients (≥0.994). LOD and LOQ were 0.034 and 0.114 μg/kg for AFB1, 0.024 and 0.082 μg/kg for AFB2, 0.067 and 0.226 μg/kg for AFG1 and 0.025 and 0.084 μg/kg for AFG2. The recovery of actual samples spiked with analytes (at 5, 15 and 20 μg/kg) were from 93.8 to 105.1%. Overall, an accurate AFs analysis method was developed and could be applied to the determination of AFs in various food and agricultural products.

A Comprehensive Review of the Analytical Development and Validation of Alprazolam in Bulk and Pharmaceutical Dosage Form

Alprazolam is approved in 2003 by US FDA. Alprazolam is available as an extended-release tablet, a mouth dissolving tablet (a tablet that dissolves rapidly in the mouth), as well as a concentrated solution (liquid). For the treatment of severe anxiety and panic disorder, alprazolam is one of the most commonly prescribed benzodiazepines. IPUAC name of Alprazolam is 8-Chloro-1-methyl-6-phenyl-4H-[1,2,4]triazolo[4,3-a] [1, 4] benzodiazepine. The molecular formula and molecular Weight is C17H12ClFN4 and 326.75 g/mol. These articles may serve as an overview of Alprazolam with their drug profile, pharmacology, pharmacokinetics, and analytical HPLC procedures that are commonly employed in determining common provision issues. These reviews cover topics such as mobile phase, mobile phase ratio, column, retention time, flow rate, UV detector wavelength, run time, and more. Linearity, percent recovery, detection limit, and quantification limit are all validating parameters. The pharmaceutical analysis serves as an internal control for pure and pharmaceutical dosage forms, which is critical for quality assurance. The development of analytical methods has emerged as a crucial study activity.

Analytical method development and validation analysis for quantitative assessment of cypermethrin by HPLC procedure

Analytical method development and validation analysis for quantitative assessment of cypermethrin by HPLC procedure

Analytical method development and validation analysis for quantitative assessment of cypermethrin by HPLC procedure

The precise, systematic, definite, particular, linear, exact and robust scientific method was developed and validated for the assay of Cypermethrin in Cypermethrin 25% EC Insecticide for Injection, USP 500 mg/vial. Presently utilized Cypermethrin 25 % EC Insecticide as a working standard having limit was 95% for method development and validation. The quantitative determination bring to accomplished by HPLC- Waters - Alliance 510 system equipped with UV/PDA detector. Methanol, Acetonitrile and water in the ratio (60:20:20 v/v/v) used as mobile phase and flow rate 1.0 ml / min. with 20 minutes run time. The detection was carried at 225 nm with Nucleosil C18 column (250 mm × 4.6 mm x 10 µm) and ambient column temperature was maintained. In this connection, method uses the 20 μl injection volume and diluent as a blank solution. The linearity of this method was found to be linear in the range of 50% to 150% of the working concentration and the range for the analytical method is 25 ppm to 75 ppm. The accuracy and precision of the method were within acceptable. The present developed HPLC method is found to be suitable. The analytical solution was found to be stable up to 48 Hrs at room temperature.

QUANTITATIVE DETERMINATION OF TERBINAFINE RESIDUES ON THE PHARMACEUTICAL MANUFACTURING EQUIPMENT SURFACES

Background: Cleaning validation is a critical analytical responsibility required by GMP (Good Manufacturing Practice), which confirms the effectiveness of the cleaning standard procedure in the pharmaceutical manufacturing process for all the produced drug products contacting the shared pharmaceutical manufacturing equipment. The need to carry out cleaning validation is especially important when the drug product is the “worst-case” for the cleaning procedure regarding solubility, therapeutic potency, toxicity, and surface adherence of active pharmaceutical ingredients (API). Estimation of API residues requires a selective and sensitive method capable of quantitative determination of API traces remaining over the surface of manufacturing equipment after the cleaning procedure. Aim: The present study demonstrates the suitability and the applicability of the proposed method obtained with a combination of a sensitive, selective, and specific analytical HPLC and effective swab wipe sampling procedures for quantitative estimation of terbinafine residues in samples collected from pharmaceutical manufacturing equipment surfaces and the efficiency of the developed standard procedure for cleaning of the shared manufacturing equipment in support of cleaning validation. Methods: The swab sampling procedure was developed to obtain a suitable recovery (>80 %). The surface (sampling area – 25 cm2) was wiped with one swab moistened with methanol. The analytical procedure was developed using the HPLC system “Agilent 1260 Infinity II” and BDS Hypersil C18 250×4.6 mm, 5 ?m column with an isocratic elution of mobile phase. Results and Discussion: The developed combined method was validated concerning chromatographic system suitability test, specificity, linearity range, accuracy, precision, the limit of detection (LOD), and quantitation (LOQ). The stability of terbinafine HCl solutions in methanol, the swab material, and membrane filter compatibility was also studied. The calibration curve was linear (R2=0.99999) over a concentration range 0.00003-0.025 mg/mL; LOD – 0.000015 mg/mL, and LOQ – 0.00003 mg/mL; The determined concentrations of terbinafine residues in test solutions were not more than 0.00090 mg/mL which are below the limit of cross-contamination of the next drug product. Conclusions: The performed analytical study using the developed method with a combination of an analytical HPLC and swab sampling procedures for estimation of terbinafine residues on surfaces of the pharmaceutical equipment used during the manufacture of Terbinafine HCl 250 mg uncoated tablets demonstrates the validity of the cleaning procedure. Other quality control laboratories can apply the proposed method to carry out cleaning validation on various drug formulations of terbinafine HCl.

PEGylated Lecithin-Chitosan Nanoparticle-Encapsulated Alphα-Terpineol for In Vitro Anticancer Effects.

The aim of this study was to fabrication PEGylated lecithin-chitosan nanoparticles (PLC-NPs) as alphα-Terpineol's (αT-PLC-NPs) delivery system and examine its anti-cancer effects. αT-PLC-NPs were synthesized by self-assembling method; after characterization, entrapment efficiency of α-T was measured by HPLC procedure. MTT test was conducted for cytotoxicity evaluation. Chick chorioallantoic membrane (CAM) and quantitative polymerase chain reaction (qPCR) analysis were used to determine the angiogenesis properties, and qPCR, flow cytometry, and acridine orange and propidium iodide (AO/PI) staining were used to evaluate the pro-apoptotic effects of αT-PLC-NPs. Finally, the anti-inflammatory and antibacterial activity of the αT-PLC-NPs was also evaluated. αT-PLC-NPs with a size of 220.8 nm, polydispersity index (PDI) of 0.3, zeta potential of +29.03 mV, and encapsulation efficiency of 82% showed higher inhibitory effect on MCF7 cells (IC50: 750 μg/mL) compared to HFF cells (above 1000 μg/mL). Decreased angiogenesis indices and embryonic growth factors in CAM assay, decreased expression of VEGF and VEGF-R genes, and decreased cell migration showed the inhibitory effect of αT-PLC-NPs on angiogenesis. Increased expression of P53, P21, and caspase9 genes, as well as the results of AO/PI staining along with increasing the number of SubG1 phase cells in flow cytometry, confirmed the pro-apoptotic effects of αT-PLC-NPs. Also, its anti-inflammatory effects were demonstrated by inhibiting the expression of pro-inflammatory cytokines (TNF-α and IL-6). The inhibitory power of αT-PLC-NPs in suppressing gram-positive and negative bacterial strains was demonstrated by disk diffusion (DD), minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) methods. PLC-NPs are a promising carrier for α-T transfer for preclinical studies.

Investigation of the chemical composition of cell cultures of DIOSCOREA CAUCASICA in connection with the production of BAS sources with cardioprotective potential

Dioscorea Caucasica is a source of various biologically active substances (BAS), e.g. saponins and polyphenols, which are known for their anti-atherosclerotic action. However, this species is in the Red List of many Russian regions. Biotechnological methods of production of callus, suspension, and root cultures in vitro can solve this problem. The research objective was to select the optimal BAS extraction parameters. The study featured in vitro cell cultures, grown according to standard methods, and extracts, obtained from these cultures using various organic solvents and extraction parameters, i.e. temperature, the ratio of raw material vs. extractant, time, etc. The best yield of the extract from callus cultures was observed when methanol was applied in a ratio of 1:10 at 40°C for 60 min. As for suspension cultures, the greatest yield was provided by isopropanol in a ratio of 1:10 at 40°C for 30 min. In case of root cultures, the most effective combination was that of isopropanol in a ratio of 1:10 at 40°C for 60 min. The root culture proved the source of the highest BAS content, namely caffeic acid, rutin, mangiferin, quercetin, and apigenin. The content of rutin was 13 and 22 times higher than that of callus and suspension cultures. TLC, HPLC, and NMR procedures demonstrated that the isopropanol extract contained such saponins as glucopyranoside, rhamnopyranoside, deltoside, protodioscin, spirosthenol A, and spirosthenol B. These substances are known for their antiatherosclerotic properties. Therefore, in v itro cell cultures of Dioscorea Caucasica can be used as raw materials for various pharmaceutical purposes, as well as in functional foods.

Use of qNMR to determine HPLC relative response factors for botanical reference standards used in pharmacopeial monographs

In modern botanical pharmacopeial monographs, one measurement of content is the quantitation of relevant constituents as marker compounds. The use of suitable reference standards (RSs) to quantify multiple compounds by HPLC is recommended in the U.S. Pharmacopeial (USP) botanical monographs. However, these substances may be expensive and difficult to develop into an RS. Surrogate RSs could be used instead of the actual constituents, provided that the relative response factors (RRFs) of each analyte to the selected surrogate RS are known. USP monographs of both Sichuan Lovage Rhizome and Dong Quai Root recognize Z-ligustilide as a major characteristic marker compound, making quantitation of Z-ligustilide and its analog(s) relevant for quality control. However, because Z-ligustilide is unstable, it is difficult to develop it into a quantitative RS. Instead, oxybenzone was selected as a surrogate external quantitative RS because of its similar chromatographic behavior to Z-ligustilide, its stability, and affordable cost. The RRF determination of Z-ligustilide to oxybenzone by the conventional HPLC procedure is challenging due to both the instability of the purified Z-ligustilide at ambient temperature and the difficulty of determining its purity. Therefore, a qNMR method was used to overcome these challenges as it enables to directly measure the mass ratio of Z-ligustilide to oxybenzone in the stock solution without the need for weighing and purity information. In the present study, RRF values of 1.01, 0.46, and 0.89 for Z-ligustilide, senkyunolide A, and ferulic acid relative to oxybenzone, respectively, were determined using the qNMR-based methodology.

Analytical method development and validation analysis for quantitative assessment of vinclozolin by HPLC procedure

The definite, systematic, precise, particular, linear, proper and robust scientific method was developed and validated for the assay of Vinclozolin in Ronilan DF fungicide. Presently utilized Vinclozolin as a working standard having limit for assay of Vinclozolin in Ronilan DF fungicide are not less than 95.0% for method development and validation. The quantitative determination bring to accomplished by HPLC- Waters - Alliance 510 system equipped with UV/PDA detector. Methanol, Phosporic Acid and water in the ratio (55:45:0.1 v/v/v) used as mobile phase and flow rate 1.0 ml / min. with 15 minutes run time. The detection was carried at 220 nm with Newcrom R1 - 100mm x 3.2mm x 3µ column and ambient column temperature was maintained. In this connection, method uses the 20 μl injection volume and diluent as a blank solution. The linearity of this method was found to be linear in the range of 50% to 150% of the working concentration and the range for the analytical Method is 25 ppm to 75 ppm. The accuracy and precision of the method were within acceptable. The present developed HPLC method is detected to be suitable. The analytical solution was detected to be stable up to 48 Hrs at room temperature.

In vitro cytotoxicity assay of Fucoidan extracted from Turbinaria conoides against cancer cell lines MCF7, A549, and normal cell line L929

The main aim of the study is to quantify the cytotoxic property of the Fucoidan extracted from the Turbinaria conoides using the MTT assay with the standard fucose. Fucoidan was extracted using the soaked water method and it was determined using the HPLC procedure the obtained Test sample Fucoidan extracted from the Turbinaria conoides and standard fucose was subjected to the cytotoxicity assay against the MCF7 Human breast cancer cell line, A549 lung cancer cell line, and L929 normal mouse fibroblast cell line. From the results it was found that the Test sample showed good IC50 value for MCF7 cell line then A549 with an increasing concentration 24 hours incubation at 37°C The IC50 for MCF7 was 115.21 µg/ml and A549 396.46µg/ml and the Fucoidan extract was checked for its cytotoxicity against the normal mouse fibroblast cell line L929, Fucoidan was found non-lethal to the L929 mouse fibroblast normal cell line. Standard fucose also gave a significant result towards MCF7 and against the L929. This indicates that the Fucoidan extracted from Tubinaria conoides shows better anticancer potential in it. Hence its application can be further extended in the pharmacological fields.

High Pressure Liquid Chromatography Identification and Quantification of Morpholine Residual Content in Cobicistat After Derivatization

A derivatization HPLC procedure has been developed to identify and enumerate morpholine (MPE) in cobicistat (CCT) pharmaceutical active substances. MPE react with acetonitrile solution of 1-Naphthyl isothiocyanate to produce stable MPE derivative. The MPE derivative produced was identified and enumerated by HPLC based procedure. The linearity range of MPE was 0.3002675–1.201070 μg/mL with good squared correlation (0.9995), and limits of detection and quantification were 0.1000 μg/mL and 0.3001 μg/mL, respectively. Low (50% of specification quantity), medium (100% of specification quantity), and high (150% of specification quantity) concentrations of MPE were added in CCT samples to evaluate recovery rate. The added recovery rate ranged from 97.9% to 100.4%. The precision value was 0.79% relative standard deviation. The developed procedure also has good selectivity and robustness. This quantitative procedure for MPE can successfully be implemented to analyse the MPE residual quantities in CCT pharmaceutical active substances.

Transglycosylation by β-mannanase TrMan5A variants and enzyme synergy for synthesis of allyl glycosides from galactomannan

Retaining β-mannanases are glycoside hydrolases (GHs) that can potentially be applied for synthesis of glycosides by catalysis of transglycosylation reactions. A novel active-site double mutant (R171K/E205D) of the catalytic module (CM) of the family GH5 Trichoderma reesei β-mannanase (TrMan5A) was expressed in Pichia pastoris and purified. TrMan5A, CM and CM-variants R171K and R171K/E205D had pH optima between pH 4.0–5.3 and showed >80 % remaining activity after incubation at 40 °C for 48 h. The enzymes were screened for transglycosylation capacity toward oligomeric and polymeric donor substrates and alcohol acceptors using mass-spectrometry. Hydrolysis and transglycosylation products were analysed by a novel HPLC procedure using an NH2 column. R171K/E205D was superior in reactions with mannotetraose and the acceptor allyl alcohol, it had twice as high propensity for transglycosylation as wild-type TrMan5A. Wild-type TrMan5A produced the highest amounts of allyl β-mannosides (with 1–3 mannosyls) from locust bean galactomannan. Applying enzyme synergy, adding the GH27 guar α-galactosidase to the reaction (to cleave off galactomannan side-groups), gave a 2.1-fold increase of allyl mannosides and simultaneously a significant production of allyl galactopyranoside, increasing overall yield of allyl glycosides 4.4-fold, from 2.2% to 9.8%. The enzymatic synthesis of reactive allyl glycosides opens up for production of novel biomaterials and glycopolymers.

On the Squalene Content of CV Chondrolia Chalkidikis and Chalkidiki (Greece) Virgin Olive Oil.

This work is a continuation of efforts to establish the nutritional profile of virgin olive oil (VOO) from cv. Chondrolia Chalkidikis and Chalkidiki and to strengthen its positioning in the global VOO landscape. VOOs produced at an industrial scale in different olive mills of the Chalkidiki (Greece) regional unit as well as VOOs obtained at the laboratory scale from drupes of different maturity stages for four consecutive harvesting years were examined for their squalene (SQ) content using both HPLC and GC procedures. The mean values of SQ were found to be 4228 (HPLC) and 4865 (GC) mg/kg oil (n = 15) and were of the same magnitude as that in VOOs from cv Koroneiki (4134 mg/kg, n = 23). Storage of VOOs in the dark at room temperature for 18 months indicated an insignificant mean SQ content loss (~2%) in comparison to a mean loss of 26% for alpha-tocopherol content. This finding strengthens our view that SQ does not act as a radical scavenger that donates hydrogen atoms to the latter. The four consecutive harvest years studied indicated a clear declining trend in VOO SQ concentration upon olive ripening. To our knowledge, this is the first systematic work concerning the SQ content of Chondrolia Chalkidikis and Chalkidiki VOOs.

In vivo brain levels of acetylcholine and 5-hydroxytryptamine after oleoylethanolamide or palmitoylethanolamide administrations are mediated by PPARα engagement.

The peroxisome proliferator-activated receptor alpha (PPARα) is a nuclear receptor that has been linked to the modulation of several physiological functions, including the sleep-wake cycle. The PPARα recognizes as endogenous ligands the lipids oleoylethanolamide (OEA) and palmitoylethanolamide (PEA), which in turn, if systemically injected, they exert wake-promoting effects. Moreover, the activation of PPARα by the administration of OEA or PEA increases the extracellular contents of neurotransmitters linked to the control of wakefulness; however, the role of PPARα activated by OEA or PEA on additional biochemicals related to waking regulation, such as acetylcholine (ACh) and 5-hydroxytryptamine (5-HT), has not been fully studied. Here, we have investigated the effects of treatments of OEA or PEA on the contents of ACh and 5-HT by using in vivo microdialysis techniques coupled to HPLC means. For this purpose, OEA or PEA were systemically injected (5, 10 or 30 mg/kg; i.p.), and the levels of ACh and 5-HT were collected from the basal forebrain, a wake-related brain area. These pharmacological treatments significantly increased the contents of ACh and 5-HT as determined by HPLC procedures. Interestingly, PPARα antagonist MK-886 (30 mg/kg; i.p.) injected before the treatments of OEA or PEA blocked these outcomes. Our data suggest that the activation of PPARα by OEA or PEA produces significant changes on ACh and 5-HT levels measured from the basal forebrain and support the conclusion that PPARα is a suitable molecular element involved in the regulation of wake-related neurotransmitters.

Development and Validation of an HPLC Method for the Quantification of Morin Flavonoid Encapsulated within PLGA Nanoparticles

Background: Morin flavonoid exerts neuroprotective effects with potential interest in neurodegenerative disorders. For this, the use of surface-modified polymeric nanoparticles loaded with morin is an interesting approach. Objective: To develop and validate an HPLC method for the quantification of morin released from a new delivery system consisting of poly lactic-co-glycolic (PLGA) nanoparticles functionalized with the dipeptide phe-phe (phenylalanine-phenylalanine) used to facilitate their access to the CNS. Method: The HPLC procedure was developed and validated with morin hydrate dissolved either in methanol (method A) or in methanol: 0.1N HCl (50:50, v/v, method B). Two new nanoparticle formulations were developed and characterized: morin-loaded PLGA nanoparticles (formulation F1), and morin-loaded PLGA phe-phe nanoparticles (formulation F2). Results: Method A was linear within the concentration range of 5-30 μg.mL-1 and, 1-30 μg mL-1 for method B. LOD and LOQ with method A were 1.23 μg.mL-1 and 3.90 μg.mL-1, respectively, and 0.481 μg.mL-1 and 1.458 μg.mL-1, respectively for method B. The average amount of phe-phe bound to formulation F2 (50 mg of NPs) was 431.33 μg. The encapsulation efficiency of morin within PLGA nanoparticles was around 80%. After functionalization, this value decreased significantly. Conclusion: Method B showed better sensitivity, accuracy and precision for the quantification of morin. The procedure used to functionalize the nanoparticles was adequate for linking the dipeptide to their surfaces, but this procedure is not adequate when encapsulating water-soluble compounds.

Repercussion of piperine on pharmacokinetics parameters of eletriptan in albino rats: Involvement of CYP3A and P-GP inhibition

Eletriptan is at the type of triptan medication suggested for the management of episodes of severe headache with or without aura. Piperine (1-piperoylpiperidine) is a pure alkaloid as well as the basic tanginess material from the pipli and peppercorn. There are signs that piperine inhibits Cytochrome P-450 enzymes and P-glycoprotein, rather than arouses, drug metabolism generally, thus increasing the bioavailability and effect of several medications. The analysis was undertaken to assess the results of Piperine on the pharmacokinetics of Eletriptan. Samples of blood were taken at different time points for example 0 (predose), 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 6.0, 8.0, 12, 24 hours post-treatment. The plasma concentration of Eletriptan was estimated with the HPLC procedure. In our study Cmax, Tmax, AUC0-24 , AUC0-8, AUC%, AUMC0-24, AUMC0-8, t1/2, MRT0-24, MRT0-8 and Volume of distribution were increased by approximately 51.76%, 7.55%, 72.93%, 84.42%, 42.72%, 93.56%, 128.82%, 19.97%, 11.74%, 23.96% and 3% respectively, where as clearance decreased by 42.10% when eletriptan co-administered with piperine. In summary, the results obtained herein imply that Piperine is improving the bioavailability of Eletriptan by strengthening the exposure (AUC) of their Eletriptan when concomitantly administered by the oral route. The Piperine improved the oral bioavailability of eletriptan by inhibiting CYP3A and P-GP in rats. This observation indicates the possibility that the combo of piperine along with other CYP3A and P-GP double substrates can also enhance bioavailability.