Journal of Pharmaceutical Analysis

Non-covalent binding analysis of sulfamethoxazole to human serum albumin:Fluorescence spectroscopy, UV-vis, FT-IR, voltammetric and molecular modeling

Praveen N. Naik, Sharanappa T. Nandibewoor, Shivamurthi A. Chimatadar

2015, (3): 143-152.

Abstract(103) PDF(0)

Abstract:
This study was designed to examine the interaction of sulfamethoxazole (SMZ) with human serum albumin(HSA). Spectroscopic analysis of the emission quenching at different temperatures revealed that the quenching mechanism of human serum albumin by SMZ was static mechanism. The binding constant values for the SMZ–HSA system were obtained to be 22,500 L/mol at 288 K, 15,600 L/mol at 298 K, and 8500 L/mol at 308 K. The distance r between donor and acceptor was evaluated according to the theory of F?ster energy transfer. The results of spectroscopic analysis and molecular modeling techniques showed that the conformation of human serum albumin had been changed in the presence of SMZ. The thermodynamic parameters, namely enthalpy change (ΔH0) -36.0 kJ/mol, entropy change (ΔS0) -41.3 J/mol K and free energy change (ΔG0) -23.7 kJ/mol, were calculated by using van’t Hoff equation. The effect of common ions on the binding of SMZ to HSA was tested.

Determination of diclofenac in pharmaceutical preparations by voltammetry and gas chromatography methods

Bilal Yilmaz, Ulvihan Ciltas

2015, (3): 153-160.

Abstract(87) PDF(0)

Abstract:
Rapid, sensitive and specific methods were developed for the determination of diclofenac in pharmaceutical preparations by linear sweep voltammetry (LSV) and gas chromatography (GC) with mass spectrometry (MS) detection. The linearity was established over the concentration range of 5–35μg/mL for LSV and 0.25–5μg/mL for GC–MS method. The intra- and inter-day relative standard deviation (RSD) was less than 4.39% and 4.62% for LSV and GC–MS, respectively. Limits of quantification (LOQ) were determined as 4.8 and 0.15μg/mL for LSV and GC–MS, respectively. No interference was found from tablet excipients at the selected assay conditions. The methods were applied for the quality control of commercial diclofenac dosage forms to quantify the drug and to check the formulation content uniformity.

Comparison of conventional and supported liquid extraction methods for the determination of sitagliptin and simvastatin in rat plasma by LC-ESI-MS/MS

B. Ramesha, N. Manjulaa, S.R. Bijargib, V.U.M. Sarmaa, P. Sita Devia

2015, (3): 161-168.

Abstract(105) PDF(0)

Abstract:
Three extraction methods were compared for their efficiency to analyze sitagliptin and simvastatin in rat plasma by LC–MS/MS, including (1) liquid–liquid extraction (LLE), (2) solid phase extraction (SPE) and (3) supported liquid extraction (SLE). Comparison of recoveries of analytes with different extraction methods revealed that SLE was the best extraction method. The detection was facilitated with ion trap-mass spectrometer by multiple reactions monitoring (MRM) in a positive ion mode with ESI. The transitions monitored were m/z 441.1→325.2 for simvastatin, 408.2→235.1 for sitagliptin and 278.1→260.1 for the IS. The lower limit of quantification (LLOQ) was 0.2 ng/mL for sitagliptin and 0.1 ng/mL for simvastatin. The effective SLE offers enhanced chromatographic selectivity, thus facilitating the potential utility of the method for routine analysis of biological samples along with pharmacokinetic studies.

Quality evaluation of synthetic quorum sensing peptides used in R & D

Frederick Verbeke, Evelien Wynendaele, Sarah Braet, Matthias DHondt, Bart De Spiegeleer

2015, (3): 169-181.

Abstract(118) PDF(2)

Abstract:
Peptides are becoming an important class of molecules in the pharmaceutical field. Closely related peptide-impurities in peptides are inherent to the synthesis approach and have demonstrated to potentially mask biomedical experimental results. Quorum sensing peptides are attracting high interest in R&D and therefore a representative set of quorum sensing peptides, with a requested purity of at least 95.0%, was evaluated for their purity and nature of related impurities. In-house quality control (QC) revealed a large discrepancy between the purity levels as stated on the supplier’s certificate of analysis and our QC results. By using our QC analysis flowchart, we demonstrated that only 44.0% of the peptides met the required purity. The main compound of one sample was even found to have a different structure compared to the desired peptide. We also found that the majority of the related impurities were lacking amino acid(s) in the desired peptide sequence. Relying on the certificates of analysis as provided by the supplier might have serious consequences for peptide research, and peptide-researchers should implement and maintain a thorough in-house QC.

Extraction, characterization and biological studies of phytochemicals from Mammea suriga

Mahesha M. Poojary, Kanivebagilu A.Vishnumurthy, Airody Vasudeva Adhikari

2015, (3): 182-189.

Abstract(125) PDF(1)

Abstract:
The present work involves extraction of phytochemicals from the root bark of a well-known Indian traditional medicinal plant, viz. Mammea suriga, with various solvents and evaluation of their in vitro antimicrobial and antioxidant activities using standard methods. The phytochemical analysis indicates the presence of some interesting secondary metabolites like flavonoids, cardiac glycosides, alkaloids, saponins and tannins in the extracts. Also, the solvent extracts displayed promising anti-microbial activity against Staphylococcus aureus, Bacillus subtilis and Cryptococcus neoformans with inhibition zone in a range of 20–33 mm. Further, results of their antioxidant screening revealed that aqueous extract (with IC50 values of 111.51±1.03 and 31.05±0.92μg/mL in total reducing power assay and DPHH radical scavenging assay, respectively) and ethanolic extract (with IC50 values of 128.00±1.01 and 33.25±0.89μg/mL in total reducing power assay and DPHH radical scavenging assay, respectively) were better antioxidants than standard ascorbic acid. Interestingly, FT-IR analysis of each extract established the presence of various biologically active functional groups in it.

Liquid chromatography-tandem mass spectrometry method for the estimation of adefovir in human plasma:Application to a pharmacokinetic study

Dipanjan Goswami, Sanjay Gurule, Arabinda Saha, Poonam Vats, Arshad Khuroo, Tausif Monif

2015, (3): 190-199.

Abstract(112) PDF(1)

Abstract:
An analytical method based on solid phase extraction was developed and validated for analysis of adefovir in human plasma. Adefovir-d4 was used as an internal standard and Synergi MAX RP80A (150 mm × 4.6 mm, 4μm) column provided the desired chromatographic separation of compounds followed by detection with mass spectrometry. The method used simple isocratic chromato-graphic condition and mass spectrometric detection in the positive ionization mode. The calibration curves were linear over the range of 0.50–42.47 ng/mL with the lower limit of quantitation validated at 0.50 ng/mL. Matrix effect was assessed by post-column infusion experiment to monitor phospholipids and post-extraction addition experiment was performed. The degree of matrix effect for adefovir was determined as 7.5%and ion-enhancement in five different lots of human plasma was 7.1%and had no impact on study samples analysis with 4.5 min run time. The intra- and inter-day precision values were within 7.7% and 7.8%, respectively, for adefovir at the lower limit of quantification level. Validated bioanalytical method was successfully applied to clinical sample analysis.

Selective extraction of dimethoate from cucumber samples by use of molecularly imprinted microspheres

Jiao-Jiao Du, Rui-Xia Gao, Hu Yu, Xiao-Jing Li, Hui Mu

2015, (3): 200-206.

Abstract(77) PDF(0)

Abstract:
Molecularly imprinted polymers for dimethoate recognition were synthesized by the precipitation polymerization technique using methyl methacrylate (MMA) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the cross-linker. The morphology, adsorption and recognition properties were investigated by scanning electron microscopy (SEM), static adsorption test, and competitive adsorption test. To obtain the best selectivity and binding performance, the synthesis and adsorption conditions of MIPs were optimized through single factor experiments. Under the optimized conditions, the resultant polymers exhibited uniform size, satisfactory binding capacity and significant selectivity. Furthermore, the imprinted polymers were successfully applied as a specific solid-phase extractants combined with high performance liquid chromatography (HPLC) for determination of dimethoate residues in the cucumber samples. The average recoveries of three spiked samples ranged from 78.5% to 87.9% with the relative standard deviations (RSDs) less than 4.4% and the limit of detection (LOD) obtained for dimethoate as low as 2.3μg/mL.

Isolation and characterization of a degradation product in leflunomide and a validated selective stability-indicating HPLC-UV method for their quantification

Balraj Saini, Gulshan Bansal

2015, (3): 207-212.

Abstract(177) PDF(2)

Abstract:
Leflunomide (LLM) is subjected to forced degradation under conditions of hydrolysis, oxidation, dry heat, and photolysis as recommended by International Conference on Harmonization guideline Q1A(R2). In total, four degradation products (I–IV) were formed under different conditions. Products I, II and IV were formed in alkaline hydrolytic, acidic hydrolytic and alkaline photolytic conditions. LLM and all degradation products were optimally resolved by gradient elution over a C18 column. The major degradation product (IV) formed in hydrolytic alkaline conditions was isolated through column chromatography. Based on its 1H NMR, IR and mass spectral data, it was characterized as a British Pharmacopoeial impurity B. The HPLC method was found to be linear, accurate, precise, sensitive, specific, rugged and robust for quantification of LLM as well as product IV. Finally, the method was applied to stability testing of the commercially available LLM tablets.

Application of analytical instruments in pharmaceutical analysis

2015, (3): 213-215.

Abstract(81) PDF(0)

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2015 Vol. 5, No. 3