Journal of Pharmaceutical Analysis

Application of analytical instruments in pharmaceutical analysis

2013, (6): Ⅰ-Ⅳ.

Abstract(71) PDF(0)

Abstract:

Pharmacokinetic study of inosiplex tablets in healthy Chinese volunteers by hyphenated HPLC and tandem MS techniques

Mo Chen, Yuan Zhang, Xiao-Ting Que, Ya Ding, Lin Yang, Ai-Dong Wen, Tai-Jun Hang

2013, (6): 387-393.

Abstract(93) PDF(1)

Abstract:
Inosiplex is a compound formulation composed of inosine and p-acetaminobenzoic acid (PABA) salt of N,N-dimethylamino-2-propanol (DIP). This study was to investigate the clinical plasma pharmacokinetic properties of DIP and PABA after single and multiple oral doses of inosiplex tablets in healthy Chinese volunteers. The established LC/MS/MS method for plasma DIP determination had a linear range of 0.02-10 mg/mL, and the HPLC method for plasma PABA determination had a linear range of 0.05-40 mg/mL. Linear pharmacokinetic characteristics were found with single oral doses of 0.5, 1.0 and 2.0 g. No obvious accumulation effects were observed for DIP and PABA.

Simultaneous determination of asenapine and valproic acid in human plasma using LC-MS/MS:Application of the method to support pharmacokinetic study

Ambavaram Vijaya Bhaskar Reddy, Nandigam Venugopal, Gajulapalle Madhavin

2013, (6): 394-401.

Abstract(44) PDF(1)

Abstract:
Combination of asenapine with valproic acid received regulatory approval for acute treatment of schizophrenia and maniac episodes of bipolar disorders. A simple LC-MS/MS method was developed and validated for simultaneous quantification of asenapine and valproic acid in human plasma. Internal standards were added to 300μL of plasma sample prior to liquid-liquid extraction using methyl tertiary butyl ether (MTBE). Chromatographic separation was achieved on Phenomenex C18 column (50 mm ? 4.6 mm, 5μm) in isocratic mode at 40 1C. The mobile phase used was 10 mM ammonium formate-acetonitrile (5:95, v/v) at a constant flow rate of 0.8 mL/min monitored on triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) mode. The injection volume used for LC-MS/MS analysis was 15μL and the run time was 2.5 min. These low run time and small injection volume suggest the high efficiency of the proposed method. The method was validated over the concentration range of 0.1-10.02 ng/mL and 10-20,000 ng/mL for asenapine and valproic acid respectively. The method recoveries of asenapine (81.33%), valproic acid (81.70%), gliclazide (78.45%) and benzoic acid (79.73) from spiked plasma samples were consistent and reproducible. The application of this method was demonstrated by a pharmacokinetic study in 8 healthy male volunteers with 5 mg asenapine and 250 mg valproic acid administration.

Analysis on fat-soluble components of sinapis semina from different habitats by GC-MS

Cheng Peng, Tao Zhang, Gang Zhao, Sicen Wang

2013, (6): 402-407.

Abstract(70) PDF(0)

Abstract:
A simple and rapid gas chromatography/mass spectrometry (GC/MS) analysis method was developed for the determination of fat-soluble parts of sinapis semina. Four major compounds were chosen as marker compounds to evaluate the method. Various extraction techniques were evaluated and the greatest efficiency was observed with sonication extraction using diethyl ether. The method was valuated as follows:acceptable apparatus suitability was obtained by testing the resolutions, tailing factors and theoretical plate number of the marker compounds; the precision and reproducibility, expressed as relative standard deviation (RSD), fell within the prescribed limits. Eight samples of sinapis semina collected from markets in Xi’an were monitored by using the method. The fingerprints of those samples were analyzed by hierarchical cluster analysis (HCA) similarity analysis. The result indicated that the combination of fingerprint and HCA could be used to analyze sinapis semina from different habitats.

Rapid, simple and stability-indicating determination of polyhexamethylene biguanide in liquid and gel-like dosage forms by liquid chromatography with diode-array detection

Markus Küsters, S?ren Beyer, Stephan Kutscher, Harald Schlesinger, Michael Gerhartz

2013, (6): 408-414.

Abstract(81) PDF(0)

Abstract:
A rapid and simple method for the determination of polyhexamethylene biguanide (polyhexanide, PHMB) in liquid and gel-like pharmaceutical formulations by means of high performance liquid chromatography coupled to diode-array detection (HPLC-DAD) was developed. Best separation was achieved using a cyanopropyl bonded phase (Agilent Zorbax Eclipse XDB-CN column 4.6 mm75 mm with particle size of 3.5 mm) as well as gradient elution consisting of acetonitrile/deionized water at a flow rate of 1.0 mL/min. The optimized and applied chromatographic conditions permitted separation of polyhexanide from interacting matrix with subsequent detection at a wavelength of 235 nm with good sensitivity. The method validation was carried out with regard to the guidelines for analytical procedures demanded by the International Conference on Harmonisation (ICH). Mean recoveries of 102% and 101% for gel-like as well as liquid preparations were obtained. Suitable repeatability as well as intermediate precision could be achieved with limits of detection r0.004 mg/mL for both formulations, equivalent to r0.004% PHMB concerning sample preparation. Determination of PHMB was accomplished without tedious sample preparation. Interacting matrix could be eliminated by the chromatographic procedure with excellent performance of system suitability. All analytical requirements were fulfilled permitting a reliable and precise determination of PHMB in pharmaceuticals. Furthermore, the developed method was applied to stability testing of pharmaceutical preparations containing PHMB.

Determination of epristeride by its quenching effect on the fluorescence of L-tryptophan

Ai-Qin Gong, Xia-Shi Zhu

2013, (6): 415-420.

Abstract(113) PDF(0)

Abstract:
A rapid, novel spectrofluorimetric method to determine epristeride (EP) in biological fluids and a pharmaceutical formulation was developed, based on the fact that fluorescence intensity of L-tryptophan could be quenched by EP in the medium of pH ? 9.0. The various factors influencing fluorescence quenching were discussed. The quenching mechanism was investigated with the quenching type, synchronous fluorescence spectra and quantum efficiency. Under the optimized conditions, fluorescence quenching value (ΔF ? FL-tryptophan-FEP-L-tryptophan) showed a good linear relationship with the EP concentration ranging from 0.4 to 12.0μg/mL. The linearity, recovery and limit of detection demonstrated that the proposed method was suitable for EP determination in biological fluids and EP tablets. The method was successfully applied to the analysis of EP in real samples and the obtained results were in good agreement with the results of the official method.

Quantitative analysis of catechins in Saraca asoca and correlation with antimicrobial activity

Amey Shirolkar, Anjum Gahlaut, Anil K. Chhillar, Rajesh Dabur

2013, (6): 421-428.

Abstract(72) PDF(0)

Abstract:
Herbal medicines are highly complex and have unknown mechanisms in diseases treatment. Saraca asoca (Roxb.), De. Wild has been recommended to treat gynecological disorders and used in several commercial polyherbal formulations. In present study, efforts have been made to explore antimicrobial activity and its co-relation with the distributions of catechins in the organs of S. asoca using targeted MS/MS. Eight extracts (cold and hot water) from four different organs of S. asoca and two drugs were prepared and antimicrobial activity was assessed by microbroth dilution assay. Quantitative and qualitative analysis of catechins in crude extracts was done by using targeted and auto-MS/MS and correlated with antimicrobial activity. (t)-Catechin and (t)-epicatechin and their biosynthesis related compound were found to be up-regulated in regenerated bark and leaves extracts. (?)-Epigallocatechin was found to be significantly higher in bark water extract as compared to others but showed low antimicrobial activity. Result showed down-regulation of (?)-epigallocatechin and up-regulation of (t)-catechin and (t)-epicatechin in the regenerated bark and leaves of S. asoca. It might be the contributing factor in the antimicrobial activity of regenerated bark and leaves of the plant. The concentration of (t)-epicatechin in processed drugs (Ashokarishta) from Baidyanath was found to be seven times higher than that of Dabur Pvt. Ltd., but no antimicrobial activity was observed, indicating the variations among the plant based drugs. This will be helpful in rational use of S. asoca parts. Furthermore, the analytical method developed is sensitive, repeatable and reliable; therefore, it is suitable for quality control of herbal drugs.

Preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus by high-speed counter-current chromatography

Xin-Ying Li, Mei Yang, Jie-Yun Huang, Xiao-Xue Yu, Min-Qian Zhao, Zhi-Kun Liang, Zhi-Sheng Xie, Xin-Jun Xu

2013, (6): 429-433.

Abstract(90) PDF(0)

Abstract:
A high-speed counter-current chromatography (HSCCC) method was successfully developed for the preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus in one step. The purity of deoxyschizandrin was 98.5%, and the structure was identified by MS, UV and NMR. This method was simple, fast, convenient and appropriate to prepare pure compound as reference substances for related research on Schisandrae Sphenantherae Fructus.

Optimization and validation of a fast RP-HPLC method for the determination of dobutamine in rat plasma:Pharmacokinetic studies in healthy rat subjects

Ramesh Thippani, Nageswara Rao Pothuraju, Nageswara Rao Ramisetti, Saida Shaik

2013, (6): 434-439.

Abstract(140) PDF(1)

Abstract:
A novel isocratic reverse phase high performance liquid chromatography (RP-HPLC) with photo diode array (PDA) detection method for the determination of dobutamine (DBT) in rat plasma was developed and validated after optimization of various chromatographic conditions and other experimental parameters. Homoveratrylamine was used as an internal standard. Methanol was used as the extracting solvent for the preparation of plasma samples. Samples were separated on a Symmetry C18 (250 mm ? 4.6 mm i.d., 5μm) analytical column. Acetonitrile and 15 mM potassium dihydrogen phosphate (pH 5.0 with 0.3%TEA) (20:80, v/v) was used. The column oven temperature was optimized at 35 1C and the flow rate was 0.8 mL/min. The detection wavelength was fixed at 230 nm for entire analysis. The calibration curve was found to be linear over the concentration range of 50-2000 ng/mL (r2 ? 0.9992). The limit of quantification (LOQ) of the method was 50 ng/mL. The % RSD values of accuracy and precision values for intra and inter days were o15%at quality control (QC) concentrations. Recovery, stability and robustness were studied within the acceptable range according to ICH guidelines. The method was efficiently applied to a pharmacokinetic study in healthy Wistar rats.

Development and validation of an RP-HPLC method for simultaneous determination of Ramipril and Amlodipine in tablets

Shi-Ying Dai, Shi-Ting Qiu, Wei Wu, Chun-Mei Fu

2013, (6): 440-446.

Abstract(129) PDF(7)

Abstract:
An RP-HPLC method for the simultaneous determination of and Amlodipine (AL)Ramipril (RP) in tablets was developed and validated by Chinese Pharmacopoeia 2010. The linearity of the proposed method was investigated in the range of 0.01-0.25 mg/mL (r2 ? 0.9998) for RP and 0.014-0.36 mg/mL (r2 ? 0.9997) for AL. The limits of detection (LOD) were 0.06μg/mL and 0.02μg/mL for RP and AL, and the limits of quantitation (LOQ) were 0.2μg/mL and 0.07μg/mL, respectively. Some major impurities and degradation products did not disturb the detection of RP and AL and the assay can thus be considered stability-indicating.

Simultaneous determination of five diterpenoid alkaloids in Herba Delphinii by HPLC/ELSD

Chao-Zhan Lin, Si-Min Xie, Chen-Chen Zhu, Zeren-Dawa Bairu, Suolang-Qimei Kangsa, Dun Zhu

2013, (6): 447-451.

Abstract(69) PDF(1)

Abstract:
A HPLC-ELSD method was developed and validated for simultaneous determination of five Hetisane-type diterpenoid alkaloids in a Tibetan traditional herbal medicine, “Gebu Dilu” (Herba Delphinii), using a Kromasil C18 column (250 mm ? 4.6 mm, 5μm) with the mobile phase consisting of acetonitrile and 0.1% triethylamine in gradient (detected by evaporative light scattering detector). The linear ranges of five compounds were determined and method validation was evaluated completely. The established method is rapid and accurate with high repeatability, and can be applied for the quality control of Herba Delphinii.

Voltammetric quantitation of nitazoxanide by glassy carbon electrode

Rajeev Jain, Rajeev Kumar Yadav, Jahangir Ahmad Rather

2013, (6): 452-455.

Abstract(76) PDF(0)

Abstract:
The present study reports voltammetric reduction of nitazoxanide in Britton-Robinson (B-R) buffer by cyclic and square-wave voltammetry at glassy carbon electrode. A versatile fully validated voltammetric method for quantitative determination of nitazoxanide in pharmaceutical formulation has been proposed. A squrewave peak current was linear over the nitazoxanide concentration in the range of 20-140 mg/mL. The limit of detection (LOD) and limit of quantification (LOQ) was calculated to be 5.23μg/mL and 17.45μg/mL, respectively.

Preparative isolation of Heteroclitin D from Kadsurae Caulis using normal-phase flash chromatography

Xiao-Xue Yu, Qian-Wen Wang, Xin-Jun Xu, Wei-Jian Lv, Ming-Qian Zhao, Zhi-Kun Liang

2013, (6): 456-459.

Abstract(62) PDF(0)

Abstract:
Heteroclitin D (H.D) was successfully isolated from Kadsurae Caulis by using flash chromatography and recrystallized by methanol, 10.2 mg of H.D was obtained from 4.86 g of crude extract, and the purity determined by HPLC was 99.4%. The structure was identified by UV, IR, MS, and NMR analysis. The fast, simple and efficient method can be applied to the preparation of reference substance of H. D.

2013 Vol. 3, No. 6