Preparation and In Vivo Evaluation of Indomethacin Loaded True Nanoemulsions

Indomethacin, a potent nonsteroidal anti-inflammatory drug, has been used in the treatment of various kinds of pains, inflammation and arthritis. However, oral administration of indomethacin produces serious gastrointestinal adverse effects. Therefore the aim of the present investigation was to evaluate the anti-inflammatory effects, skin irritation, activation energy and histopathology of indomethacin from transdermally applied true nanoemulsion. The anti-inflammatory effects of true nanoemulsions were compared with marketed Indobene® gel on carrageenan-induced paw edema in rats. Skin irritation tests were performed on Wistar rats for 14 days. The % inhibition value after 12 h application was significant for optimized formulation F6 (83) as compared to marketed Indobene® gel (P<0.01). Results of skin irritation test indicated that developed true nanoemulsion is safe for human use. The significant decrease in activation energy (1.396 kcal/mol) for indomethacin across rat skin indicated that the stratum corneum lipid bilayers were significantly disrupted (P<0.05). From these results it was concluded that the developed nanoemulsion have great potential for transdermal application of indomethacin

Skin permeation mechanism and bioavailability enhancement of celecoxib from transdermally applied nanoemulsion

<p>Abstract</p> <p>Background</p> <p>Celecoxib, a selective cyclo-oxygenase-2 inhibitor has been recommended orally for the treatment of arthritis and osteoarthritis. Long term oral administration of celecoxib produces serious gastrointestinal side effects. It is a highly lipophilic, poorly soluble drug with oral bioavailability of around 40% (Capsule). Therefore the aim of the present investigation was to assess the skin permeation mechanism and bioavailability of celecoxib by transdermally applied nanoemulsion formulation. Optimized oil-in-water nanoemulsion of celecoxib was prepared by the aqueous phase titration method. Skin permeation mechanism of celecoxib from nanoemulsion was evaluated by FTIR spectral analysis, DSC thermogram, activation energy measurement and histopathological examination. The optimized nanoemulsion was subjected to pharmacokinetic (bioavailability) studies on Wistar male rats.</p> <p>Results</p> <p>FTIR spectra and DSC thermogram of skin treated with nanoemulsion indicated that permeation occurred due to the disruption of lipid bilayers by nanoemulsion. The significant decrease in activation energy (2.373 kcal/mol) for celecoxib permeation across rat skin indicated that the stratum corneum lipid bilayers were significantly disrupted (p < 0.05). Photomicrograph of skin sample showed the disruption of lipid bilayers as distinct voids and empty spaces were visible in the epidermal region. The absorption of celecoxib through transdermally applied nanoemulsion and nanoemulsion gel resulted in 3.30 and 2.97 fold increase in bioavailability as compared to oral capsule formulation.</p> <p>Conclusion</p> <p>Results of skin permeation mechanism and pharmacokinetic studies indicated that the nanoemulsions can be successfully used as potential vehicles for enhancement of skin permeation and bioavailability of poorly soluble drugs.</p

Solubility of Gliclazide in Transcutol + Water Co-solvent Mixtures at (298.15 to 333.15) K

The aim of present investigation was to determine the mole fraction solubility of a poorly water soluble antidiabetic drug gliclazide (GLZ) in mono-solvents and various Transcutol + water co-solvent mixtures at (298.15 to 333.15) K. The experimental solubility of GLZ was measured by shake flask method and resulting data was correlated with the modified Apelblat model at each temperature studied. Good correlation was observed between the experimental data of GLZ and calculated one with absolute relative deviation in the range of (0.050 to 5.680) %. The correlation coefficients were observed in the range of 0.9966 to 0.9995 which indicated good fitting of experimental solubility data. The lowest mole fraction solubility of GLZ was observed in pure water (1.9 × 10–6 at 298.15 K) whereas the highest one was observed in pure Transcutol (11.9 × 10–3 at 298.15 K). The enthalpies and entropies for GLZ dissolution were observed as positive values in the range of (15.742 to 40.551) kJ mol–1 and (52.801 to 121.721) J mol–1 K–1, respectively in all sample matrices. These results of thermodynamic parameters indicated that the dissolution of GLZ is endothermic and an entropy-driven process. Based on current solubility data, GLZ was considered as practically insoluble (poorly soluble) in pure water and soluble in Transcutol. These preliminary studies indicated that Transcutol could be used as a co-solvent for solubility enhancement of GLZ which could help in preformulation studies and formulation development of GLZ

Simultaneous estimation of cinnamaldehyde and eugenol in essential oils and traditional and ultrasound-assisted extracts of different species of cinnamon using a sustainable/green HPTLC technique

A wide range of analytical techniques are reported for the determination of cinnamaldehyde (CCHO) and eugenol (EOH) in plant extracts and herbal formulations either alone or in combination. Nevertheless, sustainable/green analytical techniques for the estimation of CCHO and EOH either alone or in combination are scarce in the literature. Accordingly, the present research was carried out to establish a rapid, highly sensitive, and sustainable high-performance thin-layer chromatography (HPTLC) technique for the simultaneous estimation of CCHO and EOH in the traditional and ultrasound-assisted methanolic extracts of Cinnamomum zeylanicum, C. burmannii, and C. cassia and their essential oils. The simultaneous estimation of CCHO and EOH was performed through NP-18 silica gel 60 F254S HPTLC plates. The cyclohexane/ethyl acetate (90:10, v v−1) solvent system was optimized as the mobile phase for the simultaneous estimation of CCHO and EOH. The greenness score of the HPTLC technique was predicted using AGREE software. The entire analysis was carried out at a detection wavelength of 296 nm for CCHO and EOH. The sustainable HPTLC technique was observed as linear in the range 10–2000 ng band−1 for CCHO and EOH. The proposed technique was found to be highly sensitive, rapid, accurate, precise, and robust for the simultaneous estimation of CCHO and EOH. The content of CCHO in traditional methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 96.36, 118.49, and 114.18 mg g−1, respectively. However, the content of CCHO in ultrasound-assisted methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 111.57, 134.39, and 129.07 mg g−1, respectively. The content of CCHO in essential oils of C. zeylanicum, C. burmannii, and C. cassia was found to be 191.20, 214.24, and 202.09 mg g−1, respectively. The content of EOH in traditional methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 73.38, 165.41, and 109.10 mg g−1, respectively. However, the content of EOH in ultrasound-assisted methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 87.20, 218.09, and 121.85 mg g−1, respectively. The content of EOH in essential oils of C. zeylanicum, C. burmannii, and C. cassia was found to be 61.26, 79.21, and 69.02 mg g−1, respectively. The amounts of CCHO and EOH were found to be significantly higher in ultrasound-assisted extracts of all species compared to its traditional extraction and hence ultrasound extraction has been proposed as a superior technique for the extraction of CCHO and EOH. The AGREE analytical score of the present analytical technique was predicted as 0.75, suggesting excellent greenness profile of the proposed analytical technique. Based on all these observations and results, the proposed sustainable HPTLC technique can be successfully used for the simultaneous estimation of CCHO and EOH in different plant extracts and herbal products

Simultaneous determination of 6-shogaol and 6-gingerol in various ginger (Zingiber officinale Roscoe) extracts and commercial formulations using a green RP-HPTLC-densitometry method

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Various analytical methodologies have been reported for the determination of 6-shogaol (6-SHO) and 6-gingerol (6-GIN) in ginger extracts and commercial formulations. However, green analytical methods for the determination of 6-SHO and 6-GIN, either alone or in combination, have not yet been reported in literature. Hence, the present study was aimed to develop a rapid, simple, and cheaper green reversed phase high-performance thin-layer chromatography (RP-HPTLC) densitometry method for the simultaneous determination of 6-SHO and 6-GIN in the traditional and ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas. The simultaneous analysis of 6-SHO and 6-GIN was carried out via RP-18 silica gel 60 F254S HPTLC plates. The mixture of green solvents, i.e., ethanol:water (6.5:3.5 v/v) was utilized as a mobile phase for the simultaneous analysis of 6-SHO and 6-GIN. The analysis of 6-SHO and 6-GIN was performed at λmax = 200 nm for 6-SHO and 6-GIN. The densitograms of 6-SHO and 6-GIN from traditional and ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were verified by obtaining their single band at Rf = 0.36 ± 0.01 for 6-SHO and Rf = 0.53 ± 0.01 for 6-GIN, compared to standard 6-SHO and 6-GIN. The green RP-HPTLC method was found to be linear, in the range of 100–700 ng/band with R2 = 0.9988 for 6-SHO and 50–600 ng/band with R2 = 0.9995 for 6-GIN. In addition, the method was recorded as “accurate, precise, robust and sensitive” for the simultaneous quantification of 6-SHO and 6-GIN in traditional and ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas. The amount of 6-SHO in traditional extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas was obtained as 12.1, 17.9, 10.5, and 9.6 mg/g of extract, respectively. However, the amount of 6-SHO in ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were obtained as 14.6, 19.7, 11.6, and 10.7 mg/g of extract, respectively. The amount of 6-GIN in traditional extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were found as 10.2, 15.1, 7.3, and 6.9 mg/g of extract, respectively. However, the amount of 6-GIN in ultrasonication-assisted extracts of ginger rhizome, commercial ginger powder, commercial capsules, and commercial ginger teas were obtained as 12.7, 17.8, 8.8, and 7.9 mg/g of extract, respectively. Overall, the results of this study indicated that the proposed analytical technique could be effectively used for the simultaneous quantification of 6-SHO and 6-GIN in a wide range of plant extracts and commercial formulations

Development and validation of stability indicating liquid chromatographic (RP-HPLC) method for estimation of ubidecarenone in bulk drug and formulations using quality by design (QBD) approach

A novel, accurate, precise and economical stability indicating Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) method, was developed and validated for the quantitative determination of ubidecarenone (UDC) in bulk drug, UDC marketed formulation and UDC loaded cubosomes (CBMs) nanocarriers through Response surface methodology (RSM) design with three factors and three levels was performed to optimize the chromatographic variables followed by forced degradation studies of UDC were performed to detect degradation peak. RP-HPLC separation was achieved using mobile phase consisting of Acetonitrile:Tetrahydrofuran:Deionised water in the ratio 55:42:3 and a flow rate of 1.0 mL/min was optimized with a standard retention time (Rt) of 2.15 min, through experiment. The method was found linear in the concentration range of 5-100 µg/mL with a regression coefficient of 0.999. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 3.04 µg/mL and 9.11 µg/mL, respectively

5\u27-nucleotidase, oxidative stress and antioxidant status in alcohol consumers and cirrhotic patients

Uvod: Cilj istraživanja bio je izmjeriti aktivnost enzima 5\u27-nukleotidaza kod bolesnika s cirozom jetre i osoba koje uzimaju alkohol. U istraživanju se ispitivao i oksidacijski stres, antioksidansi te njihova povezanost s 5\u27-nukleotidazom. Materijali i metode: Istraživanje je provedeno u tri skupine po 25 ispitanika jednake dobi i spola: I. skupina (kontrolni ispitanici), II. skupina (osobe koje uzimaju alkohol, tj. konzumenti alkohola) i III. skupina (bolesnici s cirozom jetre). Uzorci krvi prikupljeni od ispitanika centrifugirani su kako bi se odvojila plazma za analizu 5\u27-nukleotidaze. Odvojene stanice su tri puta isprane 0,9-postotnom hladnom fiziološkom otopinom i upotrebljene za analizu glutationa, malondialdehida i superoksid-dismutaze. Rezultati: Aktivnost 5\u27-nukleotidaze u serumu bila je statistički značajno povišena kod skupine bolesnika s cirozom i skupine konzumenata alkohola. Koncentracije malondialdehida bile su također statistički značajno povišene kod bolesnika s cirozom jetre i konzumenata alkohola. Koncentracije glutationa i superoksid-dismutaze bile su statistički značajno snižene u obje skupine. Zaključak: Iz ovih rezultata može se zaključiti da je aktivnost 5\u27-nukleotidaze u serumu dosljedno viša kod bolesnika s cirozom jetre i osoba koje uzimaju alkohol. Zapažena razlika mogla bi ukazivati na opseg oštećenja jetre, oštećenja hepatobilijarnog sustava i opstrukcije jetre.Background: The present study was undertaken to determine the 5\u27-nucleotidase enzyme activity in liver cirrhotic patients and alcohol consumers. Oxidative stress, antioxidants and their association with 5\u27-nucleotidase were also investigated. Methods: The study included three groups of 25 age and sex matched subjects: group I (control), group II (alcohol consumers) and group III (cirrhotic patients). Blood samples were collected and centrifuged for separation of plasma for analysis of 5\u27-nucleotidase. Separated cells were washed thrice with 0.9% w/v cold normal saline and used for the analysis of glutathione, malondialdehyde and superoxide dismutase. Results: The activity of serum 5\u27-nucleotidase was significantly increased in both cirrhotic patients and alcohol consumers. The levels of malondialdehyde were also significantly increased in both cirrhotic patients and alcohol consumers. The levels of glutathione and superoxide dismutase were significantly decreased in both cirrhotic patients and alcohol consumers. Conclusions: Study results indicated the activity of serum 5\u27-nucleotidase to be consistently higher in cirrhotic patients and alcohol consumers. The difference recorded might be pointing to the extent of liver damage, hepatobiliary damage, and biliary stasis

Solubility of tadalafil in aqueous mixtures of Transcutol® and PEG 400 revisited: correlation, thermodynamics and preferential solvation

Article analyzing mole fraction solubilities of tadalafil (3) in aqueous mixtures of Transcutol® and PEG 400 at temperatures from 298.15 to 333.15 K following Hildebrand solubility parameters

COMPARATIVE PROFILING OF BIOMARKER PSORALEN IN ANTIOXIDANT ACTIVE EXTRACTS OF DIFFERENT SPECIES OF GENUS FICUS BY VALIDATED HPTLC METHOD

Background: A simple but sensitive HPTLC method was developed for the comparative evaluation of psoralen in antioxidant active extracts of leaves of five different species of genus Ficus (Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata and Ficus vasta). Materials and Methods: HPTLC studies were carried out using CAMAG HPTLC system on Glass-backed silica gel 60F254 HPTLC pre-coated plates using selected mobile phase toluene: methanol (9:1). The antioxidant activity was carried out, using DPPH free radical method. Results: Among all the five species of genus Ficus, F. palmata and F. carica exhibited comparatively good antioxidant activity in DPPH assay. The developed HPTLC method was found to give a compact spot for psoralen (Rf = 0.55±0.001) at 305 nm. The regression equation and r2 for psoralen was found to be Y= 4.516X+35.894 and 0.998. The quantification result revealed the presence of psoralen in only two species, F. carica (0.24%, w/w) and F. palmata (1.88%, w/w) which supported their supremacy for anti-oxidant potential over other species. The statistical analysis proved that the developed method was reproducible and selective. Conclusion: The developed method can be used as an important tool to assure the therapeutic dose of active ingredients in herbal formulations as well as for standardization and quality control of bulk drugs and in-process formulations. This method can also be employed for the further study of degradation kinetics and determination of psoralen in plasma and other biological fluids

Dizajn, razvoj i vrednovanje novih nanoemulzija za transdermalnu primjenu celekoksiba

The aim of the present study was to investigate the potential of nanoemulsion formulations for transdermal delivery of celecoxib (CXB). The in vitro skin permeation profile of optimized formulations was compared with CXB gel and nanoemulsion gel. Significant increase in the steady state flux (Jss), permeability coefficient (Kp) and enhancement ratio (Er) was observed in nanoemulsion formulations T1 and T2 (p < 0.05). The highest value of these permeability parameters was obtained in formulation T2, which consisted of 2% m/m of CXB, 10% m/m of oil phase (Sefsol 218 and Triacetin), 50% m/m of surfactant mixture (Tween-80 and Transcutol-P) and 40% m/m of water. The anti-inflammatory effects of formulation T2 showed a significant increase (p < 0.05) in inhibition after 24 h compared to CXB gel and nanoemulsion gel on carrageenean-induced paw edema in rats. These results suggested that nanoemulsions are potential vehicles for improved transdermal delivery of CXB.U radu su opisana ispitivanja nanoemulzija za transdermalnu primjenu celekoksiba (CXB). Profil permeacije kroz kožu ispitivan je in vitro i uspoređivan sa CXB gelom i nanoemulzijskim gelom. U formulacijama T1 i T2 postignuto je značajno povećanje ustaljenog fluksa (Jss), koeficijenta permeabilnosti (Kp) i povećanje omjera (Er) (p < 0.05). Najveće vrijednosti parametara permeabilnosti dobivene su u formulaciji T2 koja je sadržala 2% m/m CXB, 10% m/m uljne faze (Sefsol 218 i Triacetin), 50% m/m površinski-aktivnih tvari (Tween-80 i Transcutol-P) i 40% m/m vode. Protuupalno djelovanje formulacije T2 na edem šape štakora uzrokovan karageninom značajno je povećano (p < 0.05) poslije 24 h u usporedbi sa CXB gelom i nanoemulzijskim gelom. Rezultati ukazuju na poboljšanu isporuku celekoksiba putem nanoemulzija