Therapeutic drug monitoring of levetiracetam: Method validation using high-performance liquid chromatography-ultraviolet detector technique and usefulness in patient care setting.

Objectives: To develop and validate a modified HPLC-UV method for the estimation of serum levetiracetam levels and to assess the usefulness of serum levetiracetam estimation in epileptic patients. Materials and Methods: Modification of a previously existing HPLC-UV method was performed using liquid- liquid phase extraction and processing using reverse phase analytic HPLC-UV detector technique followed by method validation. Serum samples of patients attending our hospital's Therapeutic Drug Monitoring Outpatient Department services were analyzed for levetiracetam levels using the study method. Data of the past 6 years (2015-2020) were descriptively analyzed. Results: The modified HPLC-UV method was validated as per ICH Q2 (R1) 2005 guidelines. Usefulness of levetiracetam estimation was assessed in 1383 patients (635 children, 683 adults, 40 elderly, and 25 pregnant women). Levetiracetam levels were within the therapeutic range (TR) in 520 children, 543 young adults, 35 elderly patients, and nine pregnant women. In 112 of 232 patients with low levetiracetam levels, poor compliance was elicited. Of 641 patients on polytherapy, 446 patients had levetiracetam values within TR, whereas 29 had values above and 166 patients had values less than TR. Sodium valproate, phenytoin sodium, and carbamazepine affected levetiracetam levels when given concomitantly. Levetiracetam dose was adjusted in 61 patients with abnormal levels for better clinical response. Good seizure control was noted in 913 (82.47%) patients whose levels were within TR, whereas 136 (58.62%) patients with low levels reported an increase in seizure frequency. Conclusions: The modified HPLC-UV method is simple, rapid, efficient, and reliable for assaying serum levetiracetam.

Evaluating the antibacterial and antibiofilm potential of sulphated polysaccharides extracted from green algae Chlamydomonas reinhardtii.

AIM OF THE STUDY: In this study, antibacterial and antibiofilm potential of sulphated polysaccharides (SPs) extracted from Chlamydomonas reinhardtii (Cr) was evaluated against Neisseria mucosa, Escherichia coli, Streptococcus sp. and Bacillus subtilis. METHODS AND RESULTS: Antibacterial potential of Cr-SPs was evaluated by agar-cup diffusion, time-kill and colony-forming ability (CFU), minimum inhibitory and bactericidal concentration assays. Antibiofilm potential was evaluated by biofilm inhibition, eradication, extracellular-DNA, metabolic activity and microscopy assays. Cr-SPs at 0·5 mg ml-1 showed 34·52, 48·6, 66·1 and 55·6% reduced CFU in B. subtilis, Streptococcus, N. mucosa and E. coli respectively. Minimum inhibitory concentration of Cr-SPs was as low as 480 µg ml-1 for Streptococcus, N. mucosa and 420 µg ml-1 for B. subtilis and E. coli. At 1 mg ml-1 , Cr-SPs showed 50% biofilm inhibition, whereas 4-8 mg ml-1 showed 100% inhibition. Cr-SPs also effectively dissolved preformed biofilms. Dose-dependent reduction in extracellular DNA revealed that Cr-SPs interacts with the extra polymeric substance of the biofilm and destroys them. Light microscopy reconfirmed the above results. CONCLUSION: Cr-SPs not only inhibited biofilm formation but also effectively dissolved preformed-biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: The current study showed the promising potential of Cr-SPs as antibiofilm agents. Further validation will help in developing Cr-SPs as natural antibiotics against biofilm-causing bacteria.

DNA minor groove cleaving agents: synthesis, binding and strand cleaving properties of anthraquinone-oligopyrrolecarboxamide hybrids.

Three 9,10-anthraquinone-oligopyrrolecarboxamide hybrids have been prepared as potential DNA minor groove cleaving agents. Each conjugate was designed to contain a bis- or tris-pyrrolecarboxamide moiety related to netropsin or distamycin covalently linked to a 2-substituted anthraquinone chromophore capable of triggering photocleavage of DNA. AQ(NC)-Dist, having three pyrrole rings, is related to distamycin. AQ(NC)-Net and AQ(CN)-Net are related to netropsin; they differ only by the orientation of the amide bond between the anthraquinone and the netropsin moiety. The binding properties of these compounds to various natural DNAs have been studied by footprinting and circular dichroism. The introduction of the chromophore does not abolish the capacity of the drugs to recognize AT-rich sequences in DNA selectively. There is apparently little correlation between this property and the ability to trigger photo-induced DNA cleavage. AQ(CN)-Net is almost totally inactive in the cleavage assays whereas it manifestly binds selectively to AT-rich tracts. With AQ(NC)-Net and AQ(NC)-Dist, complete conversion of form I to form II of circular DNA is obtained. Moreover, in most cases the cleavage of DNA proved to be non-specific.