ATR-FTIR spectroscopic methods for analysis of black seed oil from alginate beads

Objective: This study aimed to use attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR) for rapid quantification of black seed oil (BSO) from alginate beads and to develop simple analysis method for in vitro release study based on turbidity measurement. Methods: Guluronic acid-rich (high-G) sodium alginate was used to encapsulate BSO with the aid of Tween 80 as a stabilizer. ATR-FTIR method was developed for quantification of encapsulation efficiency of BSO by applying Beer-Lambert law after selection of a few wave number combinations. UV-vis method based on measurement of emulsion turbidity at 600 nm was also developed to quantify BSO during the releasing from alginate beads in simulated intestine buffer. Results: ATR-FTIR method exhibited linearity in the range of 25-300 mgBSO/mlemulsion (mgBSO/mlE)with R2=0.998, RSD=8.4%, LOD=0.28 mgBSO/mlE and LOQ=0.87 mgBSO/mlE. BSO-alginate beads was found to completely encapsulate BSO with around 100.5% efficiency. UV-vis method exhibited linearity in the range of 50-300 mgBSO/mlE, R2=0.9931, RSD=1.34%, LOD=0.89 mgBSO/mlE, and LOQ=2.71 mgBSO/mlE. In addition, the method showed that total amount of BSO was released at 110 min. Conclusion: These methods are considered as a practical method for quantification of BSO for encapsulation efficacy and release. They will help to accelerate and improve routine characterization of encapsulated BSO in food and pharmaceutical technology

ATR-FTIR AND SPECTROSCOPIC METHODS FOR ANALYSIS OF BLACK SEED OIL FROM ALGINATE BEADS

Objective: This study aimed to use attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR) for rapid quantification of black seed oil (BSO) from alginate beads and to develop simple analysis method for in vitro release study based on turbidity measurement.Methods: Guluronic acid-rich (high-G) sodium alginate was used to encapsulate BSO with the aid of Tween 80 as a stabilizer. ATR-FTIR method was developed for quantification of encapsulation efficiency of BSO by applying Beer-Lambert law after selection of a few wave number combinations. UV-vis method based on measurement of emulsion turbidity at 600 nm was also developed to quantify BSO during the releasing from alginate beads in simulated intestine buffer.Results: ATR-FTIR method exhibited linearity in the range of 25-300 mgBSO/mlemulsion (mgBSO/mlE)with R2=0.998, RSD=8.4%, LOD=0.28 mgBSO/mlE and LOQ=0.87 mgBSO/mlE. BSO-alginate beads was found to completely encapsulate BSO with around 100.5% efficiency. UV-vis method exhibited linearity in the range of 50-300 mgBSO/mlE, R2=0.9931, RSD=1.34%, LOD=0.89 mgBSO/mlE, and LOQ=2.71 mgBSO/mlE. In addition, the method showed that total amount of BSO was released at 110 min.Conclusion: These methods are considered as a practical method for quantification of BSO for encapsulation efficacy and release. They will help to accelerate and improve routine characterization of encapsulated BSO in food and pharmaceutical technology

QUANTIFICATION OF NIGELLA SATIVA OIL (NSO) FROM BIODEGRADABLE PLGA NANOPARTICLES USING FTIR SPECTROSCOPY

Objective: Quantification of medicinal plant, N. sativa oil, in biodegradable nanoparticles fabricated from PLGA and chitosan was impossible due to the difficulty in the oil extraction-partition method of which the nanoparticle did not dissolve in any solvent. Methods: FTIR method was developed to quantify the loading efficiency of NSO from the intact nanoparticles without the need to solubilise the nanoparticles and extract the NSO thereafter. Beer-Lambert law was applied in the quantification following selection of a few wave number combinations. Results: The method exhibited linearity in the range NSO/PLGA=5-150% with R2=0.9911, RSD=1.68%, LOD=0.89% and LOQ=2.68%. NSO-PLGA nanoparticles revealed complete encapsulation of NSO (loading efficiency=101.5±2.2%) while chitosan-NSO-PLGA nanoparticle showed lower loading efficiency (84.5±1.7%) due to presence of the hydrophilic polymer, i. e. chitosan. This method is fast and easy to apply and does not require sample processing. The method will help to accelerate and improve routine characterisation of NSO nanoparticles during development and optimisation stage

Taste masking efficacy of paracetamol alginate beads

Many pharmaceutical drugs have unpleasant tastes, most of them being bitter. The problems of bitterness of drugs create a serious challenge to pharmacist especially when the drugs need to be taken by paediatric patients. Paracetamol is one of the common medicines that have a bitter taste. Due to this, taste masking becomes one of the important steps during formulation. Therefore, this study aims to develop a paediatric dosage form of paracetamol through using microencapsulation technique. It is a coating technique that provides a physical barrier around the drug to avoid the drug dissolution into the mouth while keeping the drug bioavailability unchanged. The biopolymer alginate is considered as a good choice for microencapsulation purpose. Alginate is a natural polysaccharide polymer isolated from brown seaweed. There is an increase interest toward the use of alginate gel in the form of beads due to non-toxicity of alginate and their ability to encapsulate drugs under very mild conditions without using high temperatures. Alginate gel beads are prepared by drop-wise of sodium alginate solution into a solution containing divalent metal ion such as calcium ion that leads to the formation of a unique egg box-like gel structure. Furthermore, extra coating of paracetamol-alginate beads with chitosan was increase more taste masking properties. Chitosan is a polysaccharide derived from chitin by alkaline deacetylation. The term of chitosan is used to describe a series of chitosan polymers with different molecular weight, viscosity and degree of acetylation. Chitosan is insoluble in at neutral and alkaline pH values but forms salt with inorganic and organic acid. in-vitro dissolution testing was used to evaluate taste masking capability, by quantifying release in simulated oral cavity conditions. From in-vitro studies, it can be concluded that the paracetamol alginate beads able to mask the bitter taste of paracetamol

Effect of shRNA mediated silencing of YB-1 protein on the expression of matrix collagenases in malignant melanoma cell in vitro

Background and Objective: YB-1 is a transcription and oncogenic factor capable of binding to DNA and RNA performing versatile functions within normal and cancer cells. Some studies reported the binding of YB-1 with a collagenases gene promoter and influencing their expression. In addition, the role of YB-1 in malignant melanoma was not elucidated. Thus, in this study, the aim was to knock down the expression of YB-1 in A375 malignant melanoma cancer cell using the shRNA approach and study its effect on cancer cell proliferation, migration, and expression of collagenases. Methods: A375 malignant melanoma cell lines were grown in standard conditions and were transfected with three plasmids containing a retroviral pGFP-V-RS vector, two of them containing targeting sequences for YB-1 mRNA. The third plasmid contained a scrambled mRNA sequence as a negative control. Expression of YB-1 was validated using immune-fluorescence staining, RT-PCR and western blotting. The cancer cell proliferation was determined using MTT assay, serial trypan blue cell counting and cell cycle flow-cytometry analysis. Expression of collagenases (MMP1, MMP8, and MMP13) was evaluated using RT-PCR and western blotting analysis. In addition, a wound-healing assay was used to assess cell migration potential. Statistical analysis was performed using one-way ANOVA test with Bonferroni post hoc analysis to compare the quantitative results among samples. Results: The established silenced cell strains (P1 and P2) had nearly 70% knockdown in the expression of YB-1. These YB-1 silenced strains had a significant cell cycle-specific reduction in cell proliferation (p < 0.05 in serial cell counting and cell cycle flow cytometry analysis, p < 0.001 in MTT assay). In addition, YB-1 silenced strains had a remarkable reduction in cell migration potential. Expression of MMP13 was significantly reduced in YB-1 silenced strains. Conclusion: YB-1 oncoprotein is a promising target in the treatment of malignant melanoma. Silencing of this protein is associated with significant anti-proliferative, anti-invasive and MMP13 insulating properties in A375 malignant melanoma cancer cell lines

Evaluation of the efficacy of a topical slimming and anti-cellulite cream containing caffeine in vivo study

INTRODUCTION: Cellulite or lipodystrophy is a major aesthetical concern among women who want to look beautiful. OBJECTIVES: The aim of this study was to evaluate the effectiveness of a topical slimming and anti-cellulite cream containing caffeine on treatment of cellulite and other various skin parameters on human volunteers. METHODS: Placebo and formulation containing 2.5% of caffeine were prepared and applied to the upper arm and abdomen of 24 healthy volunteers, twice daily in order to determine the efficacy of the formulation as anti-cellulite and slimming agent for 12 weeks. The hydration, elasticity, trans-epidermal water loss (TEWL), body fat content and circumference measurement of abdomen and upper arm of volunteers were evaluated and monitored at baseline (before application of creams), 2nd, 4th, 8th and 12th weeks of study period to measure the effect produced by the creams. RESULTS: The significant (p<0.05) reduction of fat content of upper arm and abdomen was observed after 4 weeks. The circumference measurement of upper arm was reduced significantly (p<0.05) after 8 weeks application of formulation while circumference measurement of abdomen was reduced after 12 weeks. The TEWL of abdomen values showed a significant (p<0.05) decreased immediately after 2 week of application while the TEWL of arm was reduced significantly (p<0.05) after 4 week. Skin hydration was significantly (p<0.05) increased by both creams immediately after 2 week of application. However, there was no changes was observed in skin elasticity for both creams over 12 week period study. CONCLUSION: The formulation containing caffeine was able to improve sign of cellulite, reduced circumference measurement, fat content and TEWL as well increased hydration of skin

The impact of carbamazepine crystallinity on carbamazepine-loaded microparticle formulations

Crystallinity plays a vital role in the pharmaceutical industry. It affects drug manufacturing, development processes, and the stability of pharmaceutical dosage forms. An objective of this study was to measure and analyze the carbamazepine (CBZ) crystallinity before and after formulation. Moreover, it intended to determine the extent to which the crystallinity of CBZ would affect the drug loading, the particle size, and the release of CBZ from the microparticles. The CBZ microparticles were prepared by encapsulating CBZ in ethyl cellulose (EC) polymer using a solvent evaporation method. EC was used here as a release modifier polymer and polyvinyl alcohol (PVA) as an aqueous phase stabilizer. Factorial design was used to prepare the CBZ microparticle formulations, including polymer concentration, solvent (dichloromethane, ethyl acetate), PVA concentrations factor, the homogenization time, and homogenization speed. The crystallinity of CBZ was calculated utilizing differential scanning calorimetry (DSC) thermal analysis. The crystallinity was calculated from the enthalpy of CBZ. Enthalpy was analyzed from the area under the curve peak of CBZ standard and CBZ-loaded microparticles. DSC and ATR-FTIR assessed the possible interaction between CBZ and excipients in the microparticle. The prepared CBZ microparticles showed various changes in the crystallinity rate of CBZ. The changes in the rate of CBZ crystallinity had different effects on the particle size, the drug loading, and the release of CBZ from the polymer. Statistically, all studied factors significantly affected the crystallinity of CBZ after formulation to microparticles

Comparative assessment of plasmid DNA delivery by encapsulation within or adsorbed on poly (D, L-lactide-co-glycolide) nanoparticles

Purpose: To compare the gene delivery effectiveness of plasmid DNA (pDNA) encapsulated within poly (D,L-lactide-co-glycolide) (PLGA) nanoparticles with that adsorbed on PLGA nanoparticles. Methods: PLGA nanoparticles were prepared using solvent-evaporation method. To encapsulate pDNA within the particles, it was first complexed with cetyltrimethylammonium bromide (CTAB) and then added to the oil phase during the synthesis. For the adsorption, PLGA nanoparticles were first modified with either CTAB or chitosan and then pDNA was adsorbed on the particle surface by electrostatic interaction. Results: Nanoparticles encapsulating pDNA exhibited better plasmid loading and protection with significantly lower burst release (p < 0.05) compared to that of the nanoparticles with adsorbed plasmid. Cell uptake of chitosan-modified nanoparticles by murine neuroblastoma (N2a) cells was significantly (p < 0.05) higher than that of chitosan-free nanoparticles. Nanoparticles encapsulating pDNA showed higher transfection efficiency (p < 0.05) in N2a cells. Conclusion: Encapsulation of pDNA within PLGA nanoparticles presents a potential strategy for gene delivery that is superior to pDNA adsorbed on the nanoparticle surface. In addition, encapsulation keeps the particle surface free for further modifications such as the addition of targeting ligands