Polypyrrole/carbon dot nanocomposite as an electrochemical biosensor for liquid biopsy analysis of tryptophan in the human serum of normal and breast cancer women.

Liquid biopsy analysis represents a suitable alternative analysis procedure in several cases where no tumor tissue is available or in poor patient conditions. Amino acids can play a crucial role in aiding cancer diagnosis. Monitoring of tryptophan (Trp) catabolism can aid in tracking cancer progression. Therefore, a novel nanocomposite was fabricated using overoxidized polypyrrole film doped with nano-carbon dots (nano-CDs) on the pencil graphite electrode (PGE) surface for sensitive evaluation of Trp in human serum. Using square wave voltammetry (SWV), the overoxidized polypyrrole/carbon dots/pencil graphite electrode (Ov-Ox PPy/CDs/PGE) achieved excellent electrochemical catalytic activity for evaluating Trp. The modified electrode, known as Ov-Ox PPy/CDs/PGE, demonstrated superior electrochemical catalytic activity compared to bare PGE, CDs/PGE, PPy/PGE, and PPy/CDs/PGE for evaluation of Trp. The method's excellent sensitivity was confirmed by the low limits of detection (LOD = 0.003 µmol L-1) and limit of quantitation (LOQ = 0.009 µmol L-1). The biosensor that was developed can measure tryptophan (Trp) levels in the serum of both healthy individuals and female breast cancer patients with high accuracy and sensitivity. The results indicate that there is a significant difference, as shown by the F-test, between healthy individuals and those with breast cancer. This suggests that Trp amino acid could be an essential biomarker for cancer diagnosis. Consequently, liquid biopsy analysis presents a valuable opportunity for early disease detection, particularly for cancer.

Fabrication of water compatible and biodegradable super-paramagnetic molecularly imprinted nanoparticles for selective separation of memantine from human serum prior to its quantification: An efficient and green pathway.

A novel green magnetic molecularly imprinted solid phase extraction (MMI-SPE) for separation of memantine (MEM) from complicated matrices was proposed. The nanomaterial was synthesized via crosslinking of chitosan (CHIT) with [3-(2, 3-epoxypropoxy)-propyl] trimethoxysilane (EPPTMS) in presence of MEM as a template. The nanocomposites, in all steps, were characterized by SEM, FTIR and PXRD techniques. The adsorbed drug was removed from magnetic molecular imprinted polymer (MMIP) cavity by ethanol: acetic acid (8:2, v/v) and then, coupled with sodium 1, 2-naphthoquinone-4-sulphonate (NQS) in iodine/alkaline medium to yield highly fluorescent product, after reduction with potassium borohydride (KBH4). Variables affecting extraction of MEM from imprinted sites and its fluorometric analysis were studied. The linearity was achieved over concentration range of 1.84-95.0 ng mL-1 with LOD of 0.6 ng mL-1. The method was successfully applied for determination of MEM in its pharmaceutical tablets and human serum with recoveries of 100.8 ±â€¯3.0, 97.6 ±â€¯2.9, respectively.

Highly sensitive UHPLC-DAD method for simultaneous determination of two synergistically acting antiepileptic drugs; levetiracetam and lacosamide: Application to pharmaceutical tablets and human urine.

A simple and highly sensitive ultra-high-performance liquid chromatographic-diode array (UHPLC-DAD) detection method was developed and validated for the simultaneous estimation of levetiracetam (LEV) and lacosamide (LAC). It was clinically proven that the combination of LEV and LAC exhibits a synergistic effect against refractory seizures in mice, which was the motivation for the analysis of this binary mixture both in bulk and in human urine samples. The binary mixture was resolved on a Hypersil BDS C18 analytical column, utilizing a mobile phase of 0.050 mol L-1 phosphate buffer (pH 5.60), methanol and acetonitrile in the ratio (80:10:10 v/v/v) using catechol as an internal standard. The mobile phase was pumped at a flow rate of 1.2 mL min-1 with diode array detection at 205 nm for both drugs and 270 nm for IS. Calibration curves were linear with correlation coefficient >0.9990 over the studied concentration range of 0.1-70.0 µg mL-1 for both drugs. The developed method was reproducible with low relative standard deviation values for intra- and inter-day precision (<2.0%). Both drugs were determined in bulk, pharmaceutical formulations and human urine samples without any interference from complex matrices.

Determination of donepezil in spiked rabbit plasma by high-performance liquid chromatography with fluorescence detection.

The aim of this paper is to develop sensitive, accurate, reproducible and robust RP-HPLC with fluorescence detection for estimation of donepezil (DZ) in rabbit plasma using silodosin as the internal standard (IS). The prepared samples were quantified on reversed phase column Luna C18(2) (150 × 4.6 mm i.d., 5 µm particle size) operated at room temperature using the mobile phase consisting of methanol: 0.1% acetic acid (50 : 50, v/v) at a flow rate of 1 ml min-1. The method was fully validated according to bioanalytical validation guidelines of FDA in terms of system suitability, selectivity, sensitivity, precision and stability. It was found that the increase in peak areas followed the increase of DZ concentration in the range of 2.56-200.00 ng ml-1 with LOD of 0.85 ng ml-1. The method was successfully applied for the determination of DZ in rabbit plasma using manual shaking dispersive liquid-liquid microextraction.

Micelle and inclusion complex enhanced spectrofluorimetric methods for determination of Retigabine: Application in pharmaceutical and biological analysis.

Two new, simple, selective, and highly sensitive spectrofluorimetric methods were developed and validated for the determination of the antiepileptic drug; retigabine (RTG). The first method (Method-I) depends on enhancement of the weak native fluorescence of RTG via the use of an organized medium; sodium dodecyl sulphate (SDS) in acetate buffer (pH 3.74). The second method (Method-II) depends on the enhancement of RTG weak native fluorescence through complexation with a macromolecule; beta cyclodextrin (ß-CD) in phosphate buffer (pH 3.20). A full study of different experimental parameters influencing the fluorescence intensity was carried out. In addition, a thorough investigation of the fluorescence quantum yield, fluorophore brightness and mechanism of fluorescence enhancement was performed. A seven-fold improvement in the fluorescence intensity was brought by the first method, whereas a six and half-fold enhancement of the fluorescence intensity was obtained by the second one. Linearity was achieved over wide ranges (0.05-12.5 µg mL-1) and (0.05-15 µg mL-1) with low limits of detection (LOD) of 10.6 and 14.3 ng mL-1, and limits of quantification (LOQ) of 32.0 and 43.2 ng mL-1 for (Method-I) and (Method-II), respectively. The proposed methods were validated according to ICH and US-FDA guidelines. The applicability of the proposed methods was tested for determination of RTG in its pharmaceutical dosage forms, and to study the stability of RTG under different stress conditions according to ICH guidelines including alkaline, acidic, oxidative, thermal, and photolytic stress conditions. Moreover, the high sensitivity achieved by the proposed methods permitted the determination and detection of RTG in both spiked and real rabbit plasma samples utilizing a simple protein precipitation step followed by liquid-liquid extraction method. Percentage recoveries from rabbit plasma samples were within the acceptable limits; (93.47-104.74%) and (91.33-105.70%) for (Method-I) and (Method-II), respectively.

Selective densitometric determination of four alpha-aminocephalosporins using ninhydrin reagent.

A simple, selective, and precise densitometric method for analysis of four alpha-aminocephalosporins, namely cefaclor monohydrate, cefadroxil monohydrate, cefalexin anhydrous, and cefradine anhydrous, both in bulk drugs and in formulations was developed and validated. The method employed thin-layer chromatography (TLC) aluminium sheets precoated with silica gel G 60 F(254) as the stationary phase. The solvent system consists of ethyl acetate-methanol-water with different ratios for all studied drugs (R(f) values of 0.40-0.60). The separated spots were visualized as blue to violet color after spraying with ninhydrin reagent. The linear regression analysis data for the calibration plots of all studied drugs produced a good linear relationship with correlation coefficients ranging from 0.9990 to 0.9996 and coefficients of determination ranging from 0.9986 to 0.9992 over the concentration range 2-10 microg/spot. The limits of detection and quantitation for all studied drugs ranged from 0.09 to 0.23 and from 0.27 to 0.84 microg/spot, respectively. The developed method was applied successfully for the determination of the studied drugs in their pharmaceutical dosage forms with good precision and accuracy. Also, the method can be employed as a promising stability-indicating assay.

Kinetic spectrophotometric determination of certain cephalosporins using oxidized quercetin reagent.

A simple, precise and accurate kinetic spectrophotometric method for determination of cefoperazone sodium, cefazolin sodium and ceftriaxone sodium in bulk and in pharmaceutical formulations has been developed. The method is based upon a kinetic investigation of the reaction of the drug with oxidized quercetin reagent at room temperature for a fixed time of 30 min. The decrease in absorbance after the addition of the drug was measured at 510 nm. The absorbance concentration plot was rectilinear over the range 80-400 microg mL(-1) for all studied drugs. The concentration of the studied drugs was calculated using the corresponding calibration equation for the fixed time method. The determination of the studied drugs by initial rate, variable time and rate-constant methods was feasible with the calibration equations obtained but the fixed time method has been found to be more applicable. The analytical performance of the method, in terms of accuracy and precision, was statistically validated; the results were satisfactory. The method has been successfully applied to the determination of the studied drugs in commercial pharmaceutical formulations. Statistical comparison of the results with a well established reported method showed excellent agreement and proved that there is no significant difference in the accuracy and precision.

Analysis of cephalosporin antibiotics.

A comprehensive review with 276 references for the analysis of members of an important class of drugs, cephalosporin antibiotics, is presented. The review covers most of the methods described for the analysis of these drugs in pure forms, in different pharmaceutical dosage forms and in biological fluids.

A validated spectrofluorimetric method for determination of some psychoactive drugs.

Five psychoactive drugs namely, chlorpromazine HCl, thioridazine HCl, clomipramine HCl, imipramine HCl and desipramine HCl were analyzed by a simple spectrofluorimetric method. The method is based on oxidation of the studied drugs using cerium(IV) in presence of sulphuric acid and monitoring the fluorescence of the formed cerium(III) at lambda(ex.) = 254 nm and lambda(em.) = 355 nm. All variables affecting the reaction conditions such as; cerium(IV) concentration, sulphuric acid concentration, heating time, temperature and dilution solvents were carefully studied. The effect of potential interference due to common ingredients as glucose, sucrose, lactose, citric acid and propylene glycol were investigated. A validation study of the proposed method was carried out according to USP 2002. Beer's law was obeyed for all the studied drugs in the concentration range of 0.05-1.3 microg/ml. Limits of detection range was 0.035-0.038 microg/ml and limits of quantitation of 0.116-0.125 microg/ml were obtained. The method was successfully applied for the assay of the studied drugs in pure form and in pharmaceutical dosage forms. Results were compared with official methods. The t- and F-values were calculated and compared with the theoretical values, which indicate high accuracy and good precision of the proposed method.

Spectrofluorimetric determination of acetaminophen with N-bromosuccinimide.

A simple, sensitive, and selective method for determination of acetaminophen based on its oxidation using N-bromosuccinimide (NBS) to produce a highly fluorescent product. Optimization of reaction variables was carried out concerning NBS concentration, pH, temperature, reaction time, and stability time. Under optimal analytical conditions, the fluorescent intensity was measured at lambda emission. 442 nm (excitation at lambda 330 nm). The linearity range is 120-800 ng/mL with lower detection limit of 33.6 ng/mL acetaminophen. The method was applied successfully to the determination of the compound in pharmaceutical preparations, with average recovery of 100.3 +/- 2%. The method was also applied successfully to the determination of the drug in spiked plasma samples, with an average recovery of 101.2 +/- 1%. Interference effects of some compounds, present in combination with acetaminophen, were studied and the tolerance limits of these compounds were determined.