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Introduction

During the manufacturing of sterile drugs, it is of the utmost importance to meet the minimum requirements for asepsis recommended by the legislations on good manufacturing practices-based efficient environmental monitoring.

Aims and methods

The availability of relatively simple to use matrix-assisted laser desorption ionization–time of flight mass spectromtomy (MALDI-TOF MS) devices in the last years has changed the laboratory workflows for the microbial identification, mainly in the clinical area. Thus, the objective of this work was to evaluate the suitability of the MALDI-TOF MS technique for the identification of bacteria isolated from the environment of clean rooms used in some stages of the production of a viral vaccine. Eighteen known bacterial species commonly isolated from clean rooms studied were identified by MALDI-TOF technique and by a biochemical technique (BBL Crystal® System).

Results

Performance of MALDI-TOF MS was better than biochemical technique for correct species identifications (88.89% and 38.89%, respectively) and produced less unreliable identification (5.55% and 22.22%).

Conclusion

MALDI-TOF MS can be implemented for routine identification of bacteria in a pharmaceutical quality control laboratory, but as a database-dependent system, maybe some isolated not identified by this technique must be additionally studied and, if appropriate, added to an in-house database.

Open access

The alternative system of medicines like Unani and Ayurveda is preferred worldwide nowadays due to its therapeutic efficacy, lower side effects, holistic approach, psychological dimensions, and qualitative action of weather and seasonal requirement. A simple procedure is described for the simultaneous extraction and estimation of piperlongumine and piperine in a well-known Unani polyherbal formulation using reversed-phase high-performance liquid chromatography (HPLC). The chromatography was carried out on reversed-phase C18 (250 × 4.6 mm) column with a mobile phase containing acetonitrile—water (50:50 v/v). Detection was accomplished with ultraviolet (UV) detection at λ = 325 nm. The flow rate was kept as 1.0 mL−1. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines for accuracy (94.4–105.0%), precision (0.37–2.17% RSD), and robustness (0.14–2.11% RSD). The limit of detection (LOD) values were found as 30 and 10 ng mL−1, while limit of quantification (LOQ) was 100 and 30 ng mL−1 for piperlongumine and piperine, respectively, which proved the sensitivity of the method satisfactory enough for accurate analysis of the both piperlongumine and piperine.

Open access

In this research, a novel method was developed for the matrix solid phase dispersion (MSPD) followed by high-performance liquid chromatography (HPLC) quantification of four marker constituents (vitamin C, gallic acid, rutin, and ellagic acid) in the freeze-dried pomegranate fruit juice. Various MSPD parameters like type of dispersant, sample–dispersant ratio, solvents, its volume, and time of extraction have been optimized after many trials. Furthermore, HPLC method has been developed and optimized for the analysis of all four components. The HPLC separation was achieved using a 250 × 4.6 mm column, particle size of 5 μm, C18 reverse phase column, with a mobile phase consisting of acetonitrile and 0.05% H3PO4, in gradient elution mode with a mobile phase flow rate of 1 mL/min, using ultraviolet (UV)–visible detection at 254 nm. All calibration curves showed good linear regression (r 2 ≥ 0.9925) within test ranges. The extraction recoveries of the marker constituents analyzed by MSPD methods were found as ranging from 97.5% to 103.5%. From comparing the chromatograms, validation data and other parameters like time, labor, and feasibility, we found that MSPD technique was most suitable for the analysis as compared to conventional liquid–liquid extraction technique.

Open access
Clinical and Experimental Medical Journal
Authors: Mamun-Al-Mahtab, Kabir Uddin, Salimur Rahman, Mobin Khan, Kamal, Moniruzzaman Bhuiyan and Gulzar Hussain

Abstract

Primary biliary cirrhosis (PBC) is a rare autoimmune liver disease, not infrequent in the West, but hardly reported from the East. We report a case of PBC for the first time from Bangladesh. The patient was an elderly woman presenting with vague complaints. The diagnosis was confirmed by antibody testing and histopathology, whereas other possible causes of cirrhosis were also excluded.

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Abstract

The aim of this study was to compare the histological activity index and viral load, which one response more with lamivudine, an anti-viral agent used for treating chronic hepatitis B infected patients.

Materials and methods

Following the defined case selection protocol a total of 39 CHB cases were enrolled in the study. After initial evaluation they received with lamivudine orally for 52 weeks. Repeat biopsy could be obtained in 16 cases. 16 patients were analysed in this study. All patients had a liver biopsy at the beginning of the study before starting the drug. Lamivudine was given orally at a dose of 100 mg once daily for 52 weeks. A second liver biopsy was taken for comparison at the end of week 52. Biopsy materials were stained with Haematoxylin and Eosin staining as a routine procedure. Immunohistochemical staining for both surface and core antigens by using antibodies against these antigens were done as special staining. In routine staining biopsies were scored according to Knodell's Histological Activity Index (HAI). Improvements in histological activity index were regarded in cases where Knodell's HAI improved by 2 points or more after treatment. Both Viral Load and staining intensity of each antigen were assessed by visual inspection of 300 hepatocytes randomly under light microscope.

Results

68% (11/16) patients had a reduction of hepatic necroinflammatory HAI score (Components 1 through 3) by 2 points or more at the end of 52 weeks of lamivudine therapy compared to the pretreatment values. Improvement in viral load was found in 80% (12/15) cases. Staining intensity of both viral antibodies was also decreased following treatment.

Discussion

One of the key objectives of treatment of chronic hepatitis B remains to improve hepatic histology in addition to clearing the virus. The quest continues in this regard and antivirals are potential tools at hand in achieving this goal.

Conclusion

Lamivudine causes significant improvement in viral load in the majority of patients treated for CHB.

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Interventional Medicine and Applied Science
Authors: Kamal Dua, Shakti Dhar Shukla, Terezinha de Jesus Andreoli Pinto and Philip Michael Hansbro

Considering the various limitations associated with the conventional dosage forms, nanotechnology is gaining increased attention in drug delivery particularly in respiratory medicine and research because of its advantages like targeting effects, improved pharmacotherapy, and patient compliance. This paper provides a quick snapshot about the recent trends and applications of nanotechnology to various translational and formulation scientists working on various respiratory diseases, which can help paving a new path in developing effective drug delivery system.

Open access

Summary

Andrographolide and betulinic acid are the terpenoids having potential anti-cancer activity. The cytotoxicity activity of both the drugs was carried out separately and in combination on liver cancer HepG2 cell lines. High-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) methods were developed and validated for simultaneous estimation of these two terpenoids as per the International Conference on Harmonization (ICH) guidelines, which was applied for quantification in nanoformulation. The retention time by HPLC and retardation factor by HPTLC for andrographolide and betulinic acid were found to be 2.2 and 6.6 min, and 0.24 ± 0.01 and 0.66 ± 0.01, respectively. Both the methods were validated for accuracy, precision, repeatability, robustness, limit of detection (LOD), and limit of quantitation (LOQ). The content of andrographolide and betulinic acid in nanoformulation was found to be 96.0% and 98.0% by HPLC and 96.59% and 98.33% by HPTLC, respectively, of labelled claim.

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Summary

A validated reversed-phase HPLC method has been developed for quantitative analysis of berberine in Berberis aristata fruits and in a polyherbal formulation. Separation of berberine was achieved on a C18 column with a mobile phase consisting of a 10–80% acetonitrile gradient in 0.05% aqueous orthophosphoric acid. The flow rate was 1 mL min−1. Detection was at 266 nm. A sharp, well defined peak was obtained at a retention time of 10.0 ± 0.4 min. The method was validated in accordance with ICH guidelines for accuracy, precision, robustness, and the limits of detection (LOD) and quantification (LOQ). Results from linear regression analysis were indicative of a good linear relationship (r 2 = 0.998 ± 0.0011) in a wide concentration range (5–500 μ g mL−1). LOD and LOQ were 1.5 and 5.3 μg mL−1, respectively. Satisfactory recovery results (94.6–103.1%) were obtained by the method of standard addition. Intra-day, inter-day, and intersystem precision was satisfactory, with relative standard deviation in the range 0.7–1.8%. The berberine content of fruit of Berberis aristata and the herbal formulation were 0.033% and 0.0089% (w/w), respectively. This HPLC method for quantification of berberine can be used for quality control and standardization of several crude drugs and different herbal formulations in which berberine is present as a phyto constituent.

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Summary

In this chapter, an isocratic reverse phase HPLC determination of mimosine was developed and validated in an anti-psoriatic topically applied formulation “Lajjalu”. The chromatography was performed on a C18 column with water-orthophosphoric acid (98.8:0.2, υ/υ) as a mobile phase with a pH of 3.0 at a flow rate of 1.0 mL min−1. Detection was performed at 284 nm, and a sharp peak was obtained for mimosine at a retention time of 2.62 ± 0.01 min. Linear regression analysis data for the calibration plot showed a good linear relationship between response curve and concentration in the range of 0.050–5000 ng mL−1 and the regression coefficient was 0.9998 with the linear regression equation y = 4766.8x−17726. The detection (LOD) and quantification (LOQ) limits were 10.3 and 35.6 ng mL−1, respectively. The wide linearity range, sensitivity, accuracy, short retention time, and simple mobile phase imply the method is suitable for routine quantification of mimosine with high precision and accuracy.

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Journal of Radioanalytical and Nuclear Chemistry
Authors: Alireza Khorami-Moghadam, Amir Jalilian, Kamal Yavari, Bahram Bolouri, Ali Bahrami-Samani and Mohammad Ghannadi-Maragheh

Abstract  

Antiangiogenic monoclonal antibodies in combination with therapeutic radionuclides are potential targeted therapy agents in cancer. In this study, bevacizumab was successively labeled with [166Ho]HoCl3 after conjugation with DOTA-NHS-ester with a radiochemical purity of higher than 95% (RTLC). The conjugates were purified by molecular filtration, the average number of DOTA conjugated per mAb was calculated and total concentration was determined by spectrophotometric method and the average chelate to antibody ratio (c/a) for the conjugate used in this study was 5.8:1 and protein integrity experiments (SDS-PAGE). The biodistribution studies in wild-type rats demonstrate a similar pattern to the other radiolabeled anti-vascular endothelial growth factor A (VEGF-A) immunoconjugates. 166Ho-DOTA-bevacizumab is a potential compound for therapy/imaging of VEGF-A expression in oncology.

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