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

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

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

Fast and simple gas chromatographic method for simultaneous estimation of camphor, menthol and methyl salicylate in analgesic ointment: application in stability study

A simple, rapid and sensitive gas chromatographic (GC) method with flame ionization detector (FID) has been developed and validated for simultaneous estimation of camphor, menthol and methyl salicylate (MS). Camphor, menthol and MS were separated at about 2.753, 3.206 and 3.995 min respectively on a capillary column with helium (3.3 ml/min) as carrier gas within 11 min run time. Noninterference of any peak with the peaks of interest confirms the selectivity of method. Derived quantitation limits (QL) were 0.847, 0.684 and 6.507 μg/ml for camphor, menthol and MS respectively. The linear relationship ( R2 > 0.999) between analyte concentration vs detector response was established within a range of QL to 150% of label claim concentration for each analyte. Recovery of each analyte at 50, 100 and 150% of label claim concentration levels were obtained within 99.67–101.53% establishing high accuracy of the method. The method showed acceptable precision with low relative standard deviation or RSD (0.24–1.03%) between percent recoveries for each analyte. RSD for intermediate precision (inter day analysis, analyst variation) was less than 1%. The validated method was successfully applied for quantitative determination of camphor, menthol and MS in stability samples of an analgesic ointment produced by IKOP Sdn. Bhd., Malaysia

Confocal laser scanning microscope analysis on post-biofilm assessment of biofilmproducing osteomyelitic staphylococcus aureus treated with new gentamicin-nigella sativa fusion emulsion (GNFE)

Biofilm complicates osteomyelitis as there is antibiotic resistance and toxicity involved. In order to overcome the challenges of current treatment, gentamicin, the current antibiotic of choice for osteomyelitis, is fused with Nigella sativa oil to form an emulsion. Its efficacy as anti-biofilm agent is assessed using confocal laser scanning microscope (CLSM) against different strains of biofilm-producing Staphylococcus aureus. These strains of S.aureus were first allowed to express biofilm before being exposed for 24 hours to the emulsion with (0.1% (w/v) gentamicin; 40.2% (v/v) N.sativa. Later, the emulsion was removed and the biofilm was stained with fluorescence staining. The slides were viewed under CLSM at 100 times resolution. 3D images of biofilm were reconstructed, using Image J software, to measure the thickness of biofilm and viability of bacteria cells. Results revealed that the emulsion significantly reduced biofilm thickness compared to gentamicin and N.sativa alone in all strains of S.aureus (Tukey’s test p < 0.05). The emulsion was also able to produce more than 80% and 15% surface percentage (%) of non-viable (dead) bacteria in the sensitive and resistant strain, respectively, at a significant level when compared to gentamicin and N.sativa (Tukey’s test p < 0.05). As a conclusion, this new fusion of gentamicin-N.sativa may be effective towards the biofilm of S.aureus, and can be developed further as a new promising anti-biofilm agent in osteomyelitis

Effect of surfactants on plasmid DNA stability and release from Poly (D, L-lactide-co-glycolide) microspheres

Purpose: To evaluate the effect of surfactants on plasmid DNA during preparation and release from polylactic glycolide (PLGA) microspheres. Methods: Various surfactants, both ionic and non-ionic (Span, Tween, Triton X100, cetyltrimethylammonium bromide and sodium dodecyl sulphate), were added during the microsphere preparation and their effect was evaluated. Supercoil index (SCI) was introduced as a harmonised value derived from encapsulation efficiency and supercoil preservation efficiency in order to evaluate the impact of different surfactants on pDNA encapsulation. Results: Polyvinyl alcohol and Span revealed low SCI whereas Tween increased the SCI in a fraction- dependent manner. The Tween blend of hydrophilic-lipophilic balance (HLB) of 16 and Triton X-100 (HLB = 13.5) showed the highest SCI. Span revealed high burst release of pDNA whereas Triton X-100 exhibited low burst release. Following the burst release, diffusion mechanism was found to predominate in DNA release. Conclusion: The microspheres were non-toxic to the neuro-2a cells which suggest they can be potentially used in the gene therapy of neuronal diseases