Journal of Pharmaceutical Analysis

Application of analytical instruments in pharmaceutical analysis

2013, (5): Ⅰ-IV.

Abstract(74) PDF(5)

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Fused-core particle technology in high-performance liquid chromatography:An overview

Joseph J. Kirkland, Stephanie A. Schuster, William L. Johnson, Barry E. Boyes

2013, (5): 303-312.

Abstract(125) PDF(2)

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The advent of superficially porous particles (SPPs) for packed HPLC columns has changed the way that many practitioners have approached the problem of developing needed separations. The very high efficiency of such columns, combined with convenient operating conditions, modest back pressures and the ability to use conventional HPLC instruments has resulted in intense basic studies of SPP technology, and widespread applications in many sciences. This report contains an overview of the SPP technology first developed in 2006 by Advanced Materials Technology, Inc., for sub-3-mm particles, then expanded into a family of SPP products with different particle sizes, pore sizes and other physical parameters. This approach was designed so that each particle of the family could be optimized for separating a particular group of compounds, usually based on solute size.

Kinetic performance comparison of fully and superficially porous particles with sizes ranging between 2.7 lm and 5 lm:Intrinsic evaluation and application to a pharmaceutical test compound

K. Broeckhoven, D. Cabooter, G. Desmet

2013, (5): 313-323.

Abstract(82) PDF(0)

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The reintroduction of superficially porous particles has resulted in a leap forward for the separation performance in liquid chromatography. The underlying reasons for the higher efficiency of columns packed with these particles are discussed. The performance of the newly introduced 5 mm superficially porous particles is evaluated and compared to 2.7 mm superficially porous and 3.5 and 5 mm fully porous columns using typical test compounds (alkylphenones) and a relevant pharmaceutical compound (impurity of amoxicillin). The 5 mm superficially porous particles provide a superior kinetic performance compared to both the 3.5 and 5 mm fully porous particles over the entire relevant range of separation conditions. The performance of the superficially porous particles, however, appears to depend strongly on retention and analyte properties, emphasizing the importance of comparing different columns under realistic conditions (high enough k) and using the compound of interest.

Determination of sodium hyaluronate in pharmaceutical formulations by HPLC-UV

K. Ruckmani, Saleem Z. Shaikh, Pavne Khalil, M.S. Muneera, O.A. Thusleem

2013, (5): 324-329.

Abstract(184) PDF(4)

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A liquid chromatography (HPLC) method with UV detection was developed for determination of sodium hyaluronate in pharmaceutical formulation. Sodium hyaluronate is a polymer of disaccharides, composed of D-glucuronic acid and D-N-acetylglucosamine, linked via alternating β-1, 4 and β-1, 3 glycosidic bonds. Being a polymer compound it lacks a UV absorbing chromophore. In the absence of a UV absorbing chromophore and highly polar nature of compound, the analysis becomes a major challenge. To overcome these problems a novel method for the determination of sodium hyaluronate was developed and validated based on size exclusion liquid chromatography (SEC) with UV detection. An isocratic mobile phase consisting of buffer 0.05 M potassium dihydrogen phosphate, pH adjusted to 7.0 using potassium hydroxide (10%) was used. Chromatography was carried out at 25 1C on a BioSep SEC S2000, 300 mm ? 7.8 mm column. The detection was carried out using variable wavelength UV-vis detector set at 205 nm. The compounds were eluted isocratically at a steady flow rate of 1.0 mL/min. Sodium hyaluronate retention time was about 4.9 min with an asymmetry factor of 1.93. A calibration curve was obtained from 1 to 38 g/mL (r40.9998). Within-day%RSD was 1.0 and between-day%RSD was 1.10. Specificity/selectivity experiments revealed the absence of interference from excipients, recovery from spiked samples for sodium hyaluronate was 99-102. The developed method was applied to the determination of sodium hyaluronate in pharmaceutical drug substance and product.

A critical quality parameter in quantitative fused-core chromatography:The injection volume

Jente Boonen, Matthias D’hondt, Lieselotte Veryser, Kathelijne Peremans, Christian Burvenich, Bart De Spiegeleer

2013, (5): 330-334.

Abstract(97) PDF(1)

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As part of the method development, the injection volume as a critical quality attribute in fast fused-core chromatography was evaluated. Spilanthol, a pharmaceutically interesting N-alkylamide currently under investigation in our laboratory, was chosen as the model compound. Spilanthol was dissolved in both PBS and MeOH/H2O (70/30, v/v) and subsequently analyzed using a fused-core system hereby selecting five chromatographic characteristics (retention time, area, height, theoretical plates and symmetry factor) as responses. We demonstrated that the injection volume significantly influenced both the qualitative and quantitative performance of fused-core chromatography, a phenomenon which is confounded with the nature of the used sample solvent. From 2 mL up to 100 mL injection volume with PBS as solvent, the symmetry factor decreased favorably by 20%. Moreover, the theoretical plates and the quantitative parameters (area and height) increased up to 30%. On the contrary, in this injection volume range, the theoretical plates for the methanol-based samples decreased by more than 60%, while the symmetry factor increased and the height decreased, both by 30%. The injection volume is thus a critical and often overlooked parameter in fused-core method description and validation.

Assay method for quality control and stability studies of a new antimalarial agent (CDRI 99/411)$

Kiran Khandelwal, Shakti Deep Pachauri, Sofia Zaidi, Pankaj Dwivedi, Ashok Kumar Sharma, Chandan Singh, Anil Kumar Dwivedi

2013, (5): 335-340.

Abstract(116) PDF(0)

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CDRI compound no. 99/411 is a potent 1,2,4-trioxane antimalarial candidate drug under development at our Institute. An HPLC method for determination of CDRI 99/411 with its starting material and intermediates has been developed and validated for in process quality control and stability studies. The analytical performance parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD) and lower limit of quantification (LLOQ) were determined according to International Conference on Harmonization ICH Q2(R1) guidelines. HPLC separation was achieved on a RP-select B Lichrospheres column (250 mm ? 4 mm, 5μm, Merck) using water containing 0.1%glacial acetic acid and acetonitrile as the mobile phase in a gradient elution. The eluents were monitored by a photo diode array detector at 245 and 275 nm. Based on signal to noise ratio of 3 and 10 the LOD of CDRI 99/411 was 0.55 mg/mL, while the LLOQ was 1.05 mg/mL. The calibration curves were linear in the range of 1.05-68 mg/mL. Precision of the method was determined by inter- and intra-assay variations within the acceptable range.

Application of LC-MS/MS for quantitative analysis of glucocorticoids and stimulants in biological fluids

Jamshed Haneef, Mohammad Shaharyar, Asif Husaina, Mohd Rashid, Ravinesh Mishra, Shama Parveen, Niyaz Ahmed, Manoj Pal, Deepak Kumar

2013, (5): 341-348.

Abstract(70) PDF(4)

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Liquid chromatography tandem mass chromatography (LC-MS/MS) is an important hyphenated technique for quantitative analysis of drugs in biological fluids. Because of high sensitivity and selectivity, LC-MS/MS has been used for pharmacokinetic studies, metabolites identification in the plasma and urine. This manuscript gives comprehensive analytical review, focusing on chromatographic separation approaches (column packing materials, column length and mobile phase) as well as different acquisition modes (SIM, MRM) for quantitative analysis of glucocorticoids and stimulants. This review is not meant to be exhaustive but rather to provide a general overview for detection and confirmation of target drugs using LC-MS/MS and thus useful in the doping analysis, toxicological studies as well as in pharmaceutical analysis.

Chiral separation of bavachinin in Fructus Psoraleae and rat plasma by liquid chromatography using permethylated-b-CD as a chiral selector

Jing-Jing Liu, Juan Zhang, Zi-Lin Chen

2013, (5): 349-353.

Abstract(41) PDF(1)

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A simple, sensitive and selective method of high-performance liquid chromatography (HPLC) has been successfully developed for separation of bavachinin enantiomers in Fructus Psoraleae and rat plasma. The separation and detection conditions of HPLC were optimized. Chiral bavachinin were separated with the mobile phase of methanol and water (70:30, v/v) at a flow rate of 1.0 mL/min. The linear ranges were in the range of 20-1000 mg/mL. The detection limits were tested as 4 ng/mL and 6 ng/mL for (t)-bavachinin and (à)-bavachinin, respectively. The method has been applied to analyze chiral bavachinin in rat plasma. HPLC-MS method was used to test the accuracy.

Copper interactions with DNA of chromatin and its role in neurodegenerative disorders

M.Govindaraju, H.S. Shekar, S.B.Sateesha, P.Vasudeva Raju, K.R.Sambasiva Rao, K.S.J. Rao, A.J.Rajamma

2013, (5): 354-359.

Abstract(116) PDF(0)

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In this study, we have demonstrated the conformational changes to DNA induced by abnormal interactions of copper using circular dichroism, in combination with UV-absorbance and fluorescence spectroscopy. Results confirm that binding of copper to bases of DNA in chromatin is concentration dependent. Binding efficiency of Cu2+ions to DNA is increased in proportion to the degree of unwinding of the double helix induced by denaturation. Altered B-DNA conformation will alter the integrity of DNA which may affect the normal process of DNA replication and transcription. Copper induced DNA damage in the brain may cause neurotoxicity and the neuronal cell death and is implicated in Alzheimer's disease and other neurological disorders.

A novel luminol-based chemiluminescence method for the determination of amikacin sulfate in serum by using trivalent copper-periodate complex

Yu-Fei Hu, Gong-Ke Li, Zhu-Jun Zhang

2013, (5): 360-366.

Abstract(101) PDF(0)

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A novel chemiluminescence (CL) reaction was based on the oxidizing reaction of luminol by the trivalent copper-periodate complex (K5[Cu(HIO6)2], DPC) in alkaline medium. The CL intensity could be enhanced in the presence of amikacin sulfate (AKS). A new CL method was developed for the determination of AKS by coupling with flow injection (FI) technology. Because of the distinctive oxidative effect of DPC, the luminol-based CL reaction could occur at a low concentration of 10à7 M. The relative CL intensity was proportional to the concentration of AKS in the range of 4.0 ? 10à9-4.0 ? 10à6 g/mL with the detection limit of 1.2 ? 10à9 g/mL. The relative standard deviation was 2.1% for 8.0 ? 10à9 g/mL AKS (n?9). The proposed method was successfully applied to the direct determination of AKS at the level of ng/mL in serum samples. The recovery varied from 97.0% to 106.3%. A possible mechanism of the CL reaction was discussed in detail by relating to the CL kinetic characteristics and electrochemical activities of the oxidant DPC.

In situ modified screen printed and carbon paste ion selective electrodes for potentiometric determination of naphazoline hydrochloride in its formulation

Gehad G. Mohamed, F.A. Nour El-Dien, Eman Y.Z. Frag, Marwa El-Badry Mohamed

2013, (5): 367-376.

Abstract(59) PDF(0)

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The construction and performance characteristics of new sensitive and selective in situ modified screen printed (ISPE) and carbon paste (ICPE) electrodes for determination of naphazoline hydrochloride (NPZ-HCl) have been developed. The electrodes under investigation show potentiometric response for NPZ-HCl in the concentration range from 7.0 ? 10-7 to 1.0 ? 10-2 M at 25 1C and the electrode response is independent of pH in the range of 3.1-7.9. These sensors have slope values of 59.770.6 and 59.270.2 mV decade?1 with detection limit values of 5.6 ? 10-7 and 5.9 ? 10-7 M NPZ-HCl using ISPE and ICPE, respectively. These electrodes show fast response time of 4-7 s and 5-8 s and exhibits lifetimes of 28 and 30 days for ISPE and ICPE, respectively. Selectivity for NPZ-HCl with respect to a number of interfering materials was also investigated. It was found that there is no interference from the investigated inorganic cations, anions, sugars and other pharmaceutical excipients. The proposed sensors were applied for the determination of NPZ-HCl in pharmaceutical formulation using the direct potentiometric method. It showed a mean average recovery of 100.2%and 102.6%for ISPE and ICPE, respectively. The obtained results using the proposed sensors were in good agreement with those obtained using the official method. The proposed sensors show significantly high selectivity, response time, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ) compared with other proposed methods.

Simultaneous determination of human plasma protein binding of bioactive flavonoids in Polygonum orientale by equilibrium dialysis combined with UPLC-MS/MS

Yong Huang, Hui Chen, Feng He, Zhi-Rong Zhang, Lin Zheng, Yue Liu, Yan-Yu Lan, Shang-Gao Liao, Yong-Jun Li, Yong-Lin Wang

2013, (5): 376-381.

Abstract(93) PDF(0)

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A simple and selective ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) assay was developed for the determination of the human plasma protein binding of four bioactive flavonoids (such as orientin and vitexin) in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C18 column with a mobile phase composed of 0.1%formic acid in acetonitrile and 0.1%aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITY?TQD system was operated under the multiple reaction monitoring (MRM) mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations (r40.99) of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.

A novel and rapid microbiological assay for ciprofloxacin hydrochloride

Edith Cristina Laignier Cazedey, Hérida Regina Nunes Salgado

2013, (5): 382-386.

Abstract(56) PDF(0)

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The present work reports a simple, fast and sensitive microbiological assay applying the turbidimetric method for the determination of ciprofloxacin hydrochloride (CIPRO HCl) in ophthalmic solutions. The validation method yielded good results and included excellent linearity, precision, accuracy and specificity. The bioassay is based on the inhibitory effect of CIPRO HCl upon the strain of Staphylococcus epidermidis ATCC 12228 used as the test microorganism. The results were treated statistically by analysis of variance (ANOVA) and were found to be linear (r?0.9994, in the range of 14.0-56.0 mg/mL), precise (intraday RSD%?2.06;interday RSD%?2.30) and accurate (recovery ? 99.7%). The turbidimetric assay was compared to the UV spectrophotometric and HPLC methods for the same drug. The turbidimetric bioassay described on this paper for determination of ciprofloxacin hydrochloride in ophthalmic solution is an alternative to the physicochemical methods disclosed in the literature and can be used in quality control routine.

2013 Vol. 3, No. 5