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In this research work, an isocratic, reversed-phase high-performance liquid chromatography-ultraviolet/visible detector method was developed for analysis of voriconazole standard (stock-solution) and in plasma samples. Optimization and validation of the method was carried out as per international guidelines. The method offered a simple liquid–liquid extraction technique, which exhibited best recovery of voriconazole along with fluconazole, i.e., internal standard. Different experimental conditions were tried and ultimately, the best outcomes were accomplished utilizing C-18 Perkin-Elmer® column with particulars of 150 mm length, 4.6 mm inner diameter and 5 μm particle size, protected by an RP-18 Perkin-Elmer® Pre-column guard cartridge with specifications of 10 μm particle size, 30 mm length and 4.6 mm inner diameter, utilizing mobile-phase of acetonitrile-water (ACN: H2O) in proportion of 60: 40 v/v, having a flow rate of 1.5 mL/min, and wavelength of 254 nm. All the analytes were observed to be separated in ≤7 min. A linear calibration curve was obtained at concentration range of 01–10 μg/mL of voriconazole. The correlation coefficient of voriconazole was observed to be 0.999, and average recovery (in percent) was 97.4%, whereas the relative standard deviation value was ≤2%. The lower limit of detection was 0.01 μg/mL, whereas, lower limit of quantification was 0.03 μg/mL, respectively. This developed method provided outstanding results of all validation parameters, i.e., recovery, accuracy, selectivity, precision and reproducibility. The method proposed for voriconazole analysis was applied effectively for further research investigation of voriconazole in human-plasma samples (to assess the pharmacokinetic parameters), pharmaceutical formulations and pharmacokinetic drug–drug interaction’s.

Cannabis abuse is a common public health issue and may lead to considerable adverse effects. Along with other effects, the dependence on cannabis consumption is a serious problem which has significant consequences on biochemical and clinical symptoms. This study intends to evaluate the harmful effects of the use of cannabis on thyroid hormonal levels, cardiovascular indicators, and psychotic symptoms in the included patients. This prospective multicenter study was conducted on cannabis-dependent patients with psychotic symptoms (n = 40) vs. healthy control subjects (n = 40). All participants were evaluated for psychiatric, biochemical, and cardiovascular physiological effects. Patients were selected through Diagnostic and Statistical Manual of Mental Disorders (DSM)-IV criteria and urine samples, exclusively for the evaluation of cannabis presence. Serum thyroid stimulating hormone (TSH), T3, and T4 levels were measured using the immunoassay technique. Patients were assessed for severity of depressive, schizophrenic, and manic symptoms using international ranking scales. Various quantifiable factors were also measured for the development of tolerance by cannabis. Among the patients of cannabis abuse, 47.5% were found with schizophrenia, 20% with schizoaffective symptoms, 10% with manic symptoms, and 22.5% with both manic and psychotic symptoms. In the group–group and within-group statistical analysis, the results of thyroid hormones and cardiovascular parameters were non-significant. The psychiatric assessment has shown highly significant (p < 0.001) difference of positive, negative, general psychopathology, and total scores [through Positive and Negative Syndrome Scale (PANSS) rating scales] in patients vs. the healthy control subjects. The study revealed that cannabis abuse did not significantly alter thyroid hormones and cardiovascular parameters due to the development of tolerance. However, the cannabis abuse might have a significant contributing role in the positive, negative, and manic symptoms in different psychiatric disorders.

Resistance to antimicrobial agents are rendering therapies ineffective around the globe, leading to increased mortality and treatment cost. Likewise, non-steroidal anti-inflammatory drugs (NSAIDs) and analgesics possess several side effects particularly peptic ulcer and gastrointestinal problems. Metallic nanoparticles significantly enhances the therapeutic efficacy of natural extracts owing to improved bioavailability thereby lowering the dose and side effects. The purpose of this research was to investigate the efficacy of gold nanoparticles (AuNPs). In this study, Sambucus wightiana whole plant aqueous extract was used for rapid and eco-friendly synthesis of AuNPs. They were characterized by various analytical techniques including UV–Visible spectroscopy (UV–Vis), transmission and scanning electron microscopy (TEM and SEM), energy dispersive X-ray spectroscopy (EDX), Zetasizer, X-ray diffractometer (XRD) and Fourier transform infra-red spectroscopy (FTIR). The UV–Vis spectra revealed a distinct absorption peak at 539 nm; TEM and SEM images confirmed the formation of heterogeneously dispersed AuNPs with an average area of 152.77 nm2 and width of 15.96 nm. The AuNPs showed significant inhibitory zones against Escherichia coli (25 mm), Staphylococcus epidermis (23 mm) and Salmonella enteritidis (18 mm) with MIC values 0.13, 0.11 and 0.16 mg/ml, respectively. Among fungal strains it showed highest percent inhibition against Fusarium solani (90%) and Microsporum canis (80%) with MIC values 0.02 and 0.01 mg/ml, respectively. It showed maximum anti-inflammatory activity (43.70, 48.80 and 57.08%) at 20 mg/kg dose at both early and late hours of inflammation. Likewise, in vitro models depicted concentration dependent inhibition of 5-LOX and COX-2 enzymes. AuNPs showed highest antioxidant activity (68.7% inhibition) at 1000 µg/ml, compared to ascorbic acid that showed 77.8% inhibition at the same concentration. Similarly, it exhibited significant (P ≤ 0.001) dose dependent analgesic effect with maximum inhibition (56.22%) at 20 mg/kg. In conclusion, the above findings suggest that AuNPs should be studied further in order to develop safe and effective formulations.

The Aquifoliaceae is an important family and has been used traditionally for some time. One of the members of this family is the Ilex dipyrena wall, which itself possesses a potential medicinal importance. This plant is traditionally used for the treatment of various ailments including pain, swelling, burns, and fever. The current study was designed to screen out the antioxidant and analgesic potential of this plant and to verify its traditional uses, along with its phytochemical profile. Extracts were subjected to antioxidant, analgesic, and phytochemical analysis using DPPH, chemical-induced (acetic acid and formalin) nociception models and GC-MS analysis, respectively. The leaf, bark, and fruit extracts showed significant antioxidant activity compared to that of standard. Likewise, all the extracts demonstrated significant (p &lt, 0.01) analgesic activity in a mice model. In acetic acid induced analgesia, the leaf, bark, and fruit extracts caused 51.64, 56.13 and 59.52% inhibition, respectively at a dose of 100 mg/kg while at 200 mg/kg it showed 83.01, 71.69 and 75.47% inhibition, respectively. In Formalin-induced paw-licking assay, fruit extract showed 59.42 and 64.19% inhibition at 200 mg/kg dose in the first and second phase, respectively. The GC-MS analysis revealed the presence of cathinone, phenylpropanolamine, dl-phenylephrine, amphetamine, myristic acid, and palmitic acid. Results of the study suggest that crude extracts from different parts of this plant may be a useful source for the development of novel analgesics. However, further investigation in terms of isolation of bioactive compounds and their toxicological evaluations are needed to validate the observed results.

Treatment of microbial infections and inflammatory conditions have many challenges in terms of efficacy and safety issues. Novel approaches such as nanoparticles based drug delivery system have shown promising results to solve some of these problems. The aim of this study was to exploit the efficacy of the synthesized silver nanoparticles. In this study, silver nanoparticles (AgNPs) were biosynthesized using root extract (aqueous) of Duchesnea indica. They were characterized using different techniques such as, ultraviolet–visible (UV–Vis) spectrophotometry, transmission and scanning electron microscopy (TEM and SEM), X-ray diffractometer (XRD), energy dispersive X-ray spectroscopy (EDX), fourier-transform infrared spectroscopy (FTIR) and zetasizer. The UV–Vis spectra gave a characteristic peak at 423 nm; XRD confirmed its crystalline structure; FTIR confirmed the involvement of phytochemicals in their capping and reduction; TEM images confirmed their spherical shape with average width of 20.49 nm and average area of 319.25 nm2. Various biological activities were performed on these NPs, such as antimicrobial, anti-inflammatory, analgesic and muscle relaxant, which showed significant results as follow. Among bacterial strains, Salmonella typhi (MIC: 0.01 mg/ml) and Escherichia coli (MIC: 0.01 mg/ml), while among that of fungal Microsporum canis (MIC: 0.53 mg/ml) and Alternaria alternata (MIC: 0.51 mg/ml) were most susceptible. The AgNPs showed maximum anti-inflammatory activity (46.15 and 56.85%) at 20 mg/kg after 3 and 5 h of drug administration, comparable to that of standard. In-vivo model exhibited concentration dependent inhibition of both COX-2 and 5-LOX enzymes. Similarly, it exhibited maximum analgesic activity (54.24%) at 20 mg/kg dose after 60 min. of pain induction. Furthermore, they depicted maximum muscle relaxation (P

Associations of GSTT1, GSTM1 and CYP1A1 gene variants with risk of developing oral cancer were evaluated in this study. A case-control study was conducted in Pashtun population of Khyber Pakhtunkhwa province of Pakistan in which 200 hospital based oral cancer cases and 151 population based healthy controls exposed to similar environmental conditions were included. Sociodemographic data were obtained and blood samples were collected with informed consent for analysis. GSTM1 and GSTT1 were analysed through conventional PCR method while specific RT-PCR method was used to detect CYP1A1 polymorphisms. Results were analyzed for conditional logistic regression model by SPSS version 20. The study shows that patients with either GSTM1 or GSTT1 null genotypes have significantly higher risk of oral cancer (adjusted odds (OR): (3.019 (1.861-4.898) and 3.011(1.865-4.862), respectively), which further increased when either one or both null genes were present in combination (adjusted odds (OR): (3.627 (1.981-6.642 and 9.261 (4.495-19.079), respectively). CYP1A1 rs4646903 gene variants individually showed weak association OR: 1.121 (0.717-1.752); however, in the presence of GSTM1 and/or GSTT1 null genotypes further increasing the association (adjusted odds (ORs): 4.576 (2.038-10.273), 5.593 (2.530-12.362) and 16.10 (3.854-67.260 for GSTM/GSTT null and CYP1A1 wild type, GSTM/GSTT either null and CYP1A1 variant alleles, and all 3 gene polymorphisms combinations, respectively). Our findings suggest that presence of GSTM1 and/or GSTT1 null genotypes along with variant alleles of CYP1A1 may be the risk alleles for oral cancer susceptibility in Pashtun population.

A novel liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed for the determination of rosuvastatin (rosuva) and its metabolite N-desmethyl rosuvastatin (NDM-rosuva) in human plasma using atorvastatin as internal standard. The method was validated according to international guidelines. The analytical column used was HiChrom® C18 (150 × 3.0 mm, 3 µm; Reading, UK) and the mobile phase comprised of 0.1% formic acid in acetonitrile and 0.1% formic acid in water (70:30 v/v), pumped at 300 µL/min. The precipitation of plasma proteins and extraction of analytes were carried out by a simple one-step procedure using acetonitrile. The calibration curves were linear (r2 = 0.999) over the concentration range of 0.2–20 ng/mL for rosuva and 0.1–10 ng/mL for NDM-rosuva. The lower limits of detection and quantification for rosuva were 0.1 and 0.2 ng/mL, whereas for NDM-rosuva, these were 0.03 and 0.1 ng/mL, respectively. The intra- and inter-day precisions expressed as relative standard deviation...

A method was developed for the determination of omeprazole, 5-hydroxy-omeprazole, and omeprazole-sulphone in human plasma. The mobile phase with 0.1% formic acid in acetonitrile and 0.1% formic acid in water (40/60 v/v) was used successfully for the first time for the analysis of these analytes. Analysis was carried out using Hichrom RP18 (150 × 3.0 mm, 3 µm. UK) column interfaced with a LTQ Orbitrap mass spectrometer. The calibration range for omeprazole and omeprazole sulphone was 0.002–1 µg/mL, whereas, for 5-hydroxy omeprazole was 0.005–1 µg/mL. The extracted ion ranges ([M + H]+) were; m/z 346.12–346.13 for omeprazole, 362.11–362.12 for 5-hydroxy-omeprazole, and omeprazole-sulphone and 384.07–384.09 for pantoprazole. The regression equation for omeprazole was 0.071x + 0.445, for 5-hydroxy omeprazole was y = 0.08x + 0.017 and for omeprazole sulphone y = 0.007x + 0.05. This method was successfully applied for the determination of pharmacokinetic parameters of omeprazole and its metabolites in human vol...

The objective of current work was to develop and evaluate thermoreversible subcutaneous drug delivery system for diclofenac sodium. The poloxamer 407, methyl cellulose, hydroxypropyl methyl cellulose and polyethylene glycol were used alone and in combination in different ratios to design the delivery system. The physical properties like Tsol-gel, viscosity, clarity of solution and gel were evaluated. The in vitro release of the drug delivery system was evaluated using membrane less method and the drug release kinetics and mechanism was predicted by applying various mathematical models to the in vitro dissolution data. Rabbits were used as in vivo model following subcutaneous injection to predict various pharmacokinetics parameters by applying Pk-Summit software. The in vitro and in vivo data revealed that the system consisting of the poloxamer 407 in concentration of 20% (DP20) was the most capable formulation for extending the drug release and maintaining therapeutic blood level of DS for longer duration (144 h). The data obtained for drug content after autoclaving the solutions indicate that autoclaving results in 6% degradation of DS. The data also suggested that the studied polymers poloxamer, MC and PG are good candidate to extend the drug release possessing a unique thermoreversible property.

The purpose of this study was to evaluate associations of the CYP1A1 gene variant rs4646903 polymorphism with the risk of developing esophageal cancer (EC). A case-control study was carried out in Pashtun population of Khyber Pakhtunkhwa province of Pakistan in which 140 hospital based EC cases and 196 population based healthy controls exposed to similar environmental conditions were included. A specific method based on the real time polymerase chain reaction (RT-PCR) was used to detect genotypes in case and control groups and results were then analyzed with SPSS version 20. In our population, individuals with CC and TC genotypes of the CYP1A1 rs4646903 polymorphism had significantly higher risk of EC (adjusted odds (OR): 15.709, 95%CI: 6.065-40.686, OR: 3.256 95%CI: 1.902-5.574 respectively). The 'C' allele was strongly associated with the disease (p0.0001). Adjusted OR was higher (1.5 times in C/C) in case of variant alleles that show the contribution of environmental and nutritional factors towards the development of EC. Our findings suggest that presence of the 'C' allele of rs4646903 (TC) may be one of the risk alleles for EC susceptibility in Pashtun population.

Smilax china Linn. is extensively used in traditional Chinese medicine (TCM) as well as in Pakistan for several medicinal purposes including their use in inflammatory disorders.The aims of the current study were to validate and assess the folk use of Smilax china Linn. on pharmacological grounds using the isolated compound at molecular, in vivo and computational levels.Seiboldogenin was isolated from ethyl acetate fraction of the plant crude extract. In vitro lipoxygenase and in vivo carrageenan-induced hind paw oedema models were used in experimental studies while molecular docking technique was used to conduct computational study.Sieboldogenin showed significant lipoxygenase inhibition (IC50: 38 microM). It also exhibited significant inhibition (p0.05) of carrageenan-induced hind paw oedema at the doses of 10 and 50mg/kg. Computational molecular docking showed its molecular interaction with important amino acid residues in the catalytic site of lipoxygenase, revealing its potential binding mode at molecular level.Sieboldogenin seems to be a potential new anti-inflammatory compound responsible for anti-inflammatory activities of Smilax china Linn. Its in vitro and in vivo inflammatory activities are in good agreement with the folk medicinal use of Smilax china Linn. in inflammatory disorders.

The objective of the current work was to develop and evaluate thermoreversible subcutaneous drug delivery system for Insulin. Thermoreversible in-situ gel system was developed and evaluated both in-vitro and in-vivo comprising of pluronic F-127 alone or in combination with methylcellulose in different ratios. The drug release kinetics and mechanism was predicted by applying various mathematical models to the in-vitro dissolution data. Rabbits were used as animal model following subcutaneous injection to predict various pharmacokinetic parameters by applying Pk-Summit® software. The in-vitro and in-vivo data revealed that the formulation IPM 15/3 consisting of the pluronic F-127 (15% w/v) and methylcellulose (3% w/v) was the most robust and capable formulation for extending the drug release and maintaining basal plasma insulin level between 10 and 40 µU/ml for 240 h (10 d).

A highly sensitive and rapid liquid chromatography–tandem mass spectrometry method was developed and validated for the determination of artemether and its metabolite dihydroartemisinin in human plasma using artemisinin as an internal standard. Chromatographic separation was achieved on a Supelco Discovery HS C18 RP column (150 mm × 4.6 mm, 5 µm; Bellefonte, USA) using acetonitrile and water with 0.1 % formic acid (80:20, v/v) as a mobile phase in isocratic mode. A high-resolution Thermo Electron Corporation LTQ-Orbitrap mass spectrometer (San Jose, USA) was used in single ion monitoring (SIM) mode using atmospheric pressure chemical ionization (APCI) as an interface. The following extracted ion ranges ([M + H]+) were monitored: m/z 267.14–267.16 for artemether, m/z 221.16–221.18 for dihydroartemisinin and m/z 283.14–283.16 for internal standard. The limit of detection (LOD) and limit of quantification (LOQ) for artemether were 0.3 and 0.8 ng mL−1, while for dihydroartemisinin were 0.2 and 0.6 ng mL−1, respectively. The validated method was successfully applied to the quantification of artemether and dihydroartemisinin in plasma samples of healthy volunteers participating in pharmacokinetic drug–food interaction studies.