Melinda Andrási

Application of capillary zone electrophoresis to the analysis and to a stability study of cephalosporins

The applicability of capillary zone electrophoresis (CZE) for the determination of cephalosporin antibiotics has been studied. In the case of the separation conditions optimised for fourteen cephalosporins, the precision of migration times was smaller than 1.3% RSD, and the values of the limit of detection ranged between 0.42 and 1.62 microg/ml. The proposed CZE method was applied to study the stability of cephalosporins in water at different temperatures (+25, +4 and -18 degrees C). It was established that the degradation of most cephalosporins was not higher than 20% at room temperature within 4 h of dissolution of these antibiotics.

Analysis and stability study of temozolomide using capillary electrophoresis

The applicability of micellar electrokinetic capillary chromatography (MEKC) for the analysis of temozolomide (TMZ) and its degradants, 3-methyl-(triazen-1-yl)imidazole-4-carboxamide (MTIC) and 5-amino-imidazole-4-carboxamide (AIC) has been studied. Using short-end injection, the analysis of TMZ and its degradants could be performed within 1.2 min. The obtained precision of migration times was better than 1.6 RSD%, and the limit of quantitation (LOQ) was 0.31-0.93 microg/mL. The therapeutic concentration of TMZ in blood samples can be determined after direct sample injection and conventional on-capillary UV detection. The proposed MEKC method was applied to study the stability of TMZ in water and serum at different pH values. It was established that the half-life of the TMZ in vitro serum at room temperature was 33 min, close to the half-life (28 min) obtained in water at pH 7.9.

Role of pathogenic oral flora in postoperative pneumonia following brain surgery

BackgroundPost-operative pulmonary infection often appears to result from aspiration of pathogens colonizing the oral cavity. It was hypothesized that impaired periodontal status and pathogenic oral bacteria significantly contribute to development of aspiration pneumonia following neurosurgical operations. Further, the prophylactic effects of a single dose preoperative cefazolin on the oral bacteria were investigated.MethodsA matched cohort of 18 patients without postoperative lung complications was compared to 5 patients who developed pneumonia within 48 hours after brain surgery. Patients waiting for elective operation of a single brain tumor underwent dental examination and saliva collection before surgery. Bacteria from saliva cultures were isolated and periodontal disease was scored according to type and severity. Patients received 15 mg/kg cefazolin intravenously at the beginning of surgery. Serum, saliva and bronchial secretion were collected promptly after the operation. The minimal inhibitory concentrations of cefazolin regarding the isolated bacteria were determined. The actual antibiotic concentrations in serum, saliva and bronchial secretion were measured by capillary electrophoresis upon completion of surgery. Bacteria were isolated again from the sputum of postoperative pneumonia patients.ResultsThe number and severity of coexisting periodontal diseases were significantly greater in patients with postoperative pneumonia in comparison to the control group (p = 0.031 and p = 0.002, respectively). The relative risk of developing postoperative pneumonia in high periodontal score patients was 3.5 greater than in patients who had low periodontal score (p < 0.0001). Cefazolin concentration in saliva and bronchial secretion remained below detectable levels in every patient.ConclusionPresence of multiple periodontal diseases and pathogenic bacteria in the saliva are important predisposing factors of postoperative aspiration pneumonia in patients after brain surgery. The low penetration rate of cefazolin into the saliva indicates that its prophylactic administration may not be sufficient to prevent postoperative aspiration pneumonia. Our study suggests that dental examination may be warranted in order to identify patients at high risk of developing postoperative respiratory infections.

Effectiveness of cephalosporins in the sputum of patients with nosocomial bronchopneumonia

ABSTRACT Nosocomial bronchopneumonia is a frequent complication in patients with chronic intratracheal intubation. Despite targeted antibiotic treatment, production of abundant bronchial secretion containing pathogen bacteria often tends to be chronic, and so mortality drastically increases. This problem led to an investigation of the penetration of five cephalosporin antibiotics into the sputum. Serum and sputum were collected from 24 chronically intubated patients having purulent nosocomial bronchopneumonia treated in an intensive care unit (ICU). Patients received the following doses intravenously every 24 h: five received 70 mg/kg of body weight cefuroxime, four received 110 mg/kg cefamandole, six received 80 mg/kg ceftriaxone, four received 80 mg/kg ceftazidime, and five received 80 mg/kg cefepime. Antibiotic concentrations in the serum and sputum were evaluated by capillary electrophoresis. MICs were determined for bacteria isolated from the purulent bronchial secretions. The mean levels of the cephalosporins in the sputum did not reach the MICs for the bacteria isolated from the same samples. Ceftriaxone was the only one of the investigated five cephalosporins that had a measurable concentration in the sputum (1.4 ± 1.2 mg/liter). The low concentration of antibiotics in the purulent tracheobronchial secretion can be one of the many reasons for ineffective therapy of nosocomial bronchopneumonia in intubated patients in the ICUs. In the case of intubated or mechanically ventilated patients having chronic bronchopneumonia, determination of drug concentration in the bronchial secretion might be considered when selecting an antibiotic for treatment.

Capillary electrophoresis for the direct determination of cephalosporins in clinical samples

SummaryThe possibility of determination of four cephalosporin antibiotics in clinical samples by capillary electrophoresis has been investigated. The separation conditions for capillary zone electrophoresis (CZE) were studied in detail. The precision of migration times measured by use of the optimized method was satisfactory (RSD<1%) and response was linearly dependent on concentration over the approximate range 2–150 mg L−1 for all the compounds studied (cefuroxime, cefotaxime, ceftriaxone, and ceftazidime). Complete separation could be achieved within 5 min. The CZE method was found to be highly suitable for direct determination of the antibiotics in clinical samples such as wound drainage, cerebrospinal fluid, and urine; for serum, however, the use of micellar electrokinetic capillary chromatography (MECC) was more advantageous.

Determination of temozolomide in serum and brain tumor with micellar electrokinetic capillary chromatography.

Micellar electrokinetic capillary chromatographic (MEKC) with photodiode-array detection was applied to determine temozolomide (TMZ) in human serum and brain tumor. The limit of quantitation (LOQ) was 0.096 μg/mL using 325 nm as detection wavelength. The method made possible that the TMZ could be detected in in vivo serum samples without sample pretreatment. In order to detect TMZ at lower concentration, an extraction with ethyl acetate was applied to preconcentrate the analyte. Small amount of brain tumor tissues (less than 1g) were lyophilized and pretreated using extraction as a clean up and concentrating step. After removing the organic solvent a final sample volume of only 10 μL was analyzed. The obtained peak concentrations (8.2-10.1 μg/mL) and T(max) (44-65 min) of TMZ in serum were similar to the data reported by others, the in vivo TMZ concentrations found in brain tumor ranged between 0.061 and 0.117 μg/g.

The Role of Equilibrium and Kinetic Properties in the Dissociation of Gd[DTPA‐bis(methylamide)] (Omniscan) at near to Physiological Conditions

[Gd(DTPA-BMA)] is the principal constituent of Omniscan, a magnetic resonance imaging (MRI) contrast agent. In body fluids, endogenous ions (Zn(2+), Cu(2+), and Ca(2+)) may displace the Gd(3+). To assess the extent of displacement at equilibrium, the stability constants of DTPA-BMA(3-) complexes of Gd(3+), Ca(2+), Zn(2+), and Cu(2+) have been determined at 37 °C in 0.15 M NaCl. The order of these stability constants is as follows: GdL≈CuL>ZnL≫CaL. Applying a simplified blood plasma model, the extent of dissociation of Omniscan (0.35 mM [Gd(DTPA-BMA)]) was found to be 17% by the formation of Gd(PO4), [Zn(DTPA-BMA)](-) (2.4%), [Cu(DTPA-BMA)](-) (0.2%), and [Ca(DTPA-BMA)](-) (17.7%). By capillary electrophoresis, the formation of [Ca(DTPA-BMA)](-) has been detected in human serum spiked with [Gd(DTPA-BMA)] (2.0 mM) at pH 7.4. Transmetallation reactions between [Gd(DTPA-BMA)] and Cu(2+) at 37 °C in the presence of citrate, phosphate, and bicarbonate ions occur by dissociation of the complex assisted by the endogenous ligands. At physiological concentrations of citrate, phosphate, and bicarbonate ions, the half-life of dissociation of [Gd(DTPA-BMA)] was calculated to be 9.3 h at pH 7.4. Considering the rates of distribution and dissociation of [Gd(DTPA-BMA)] in the extracellular space of the body, an open two-compartment model has been developed, which allows prediction of the extent of dissociation of the Gd(III) complex in body fluids depending on the rate of elimination of the contrast agent.

Determination of gadolinium-based magnetic resonance imaging contrast agents by micellar electrokinetic capillary chromatography

MEKC with DAD was applied to detect six Gd‐based contrasting agents (CAs) (Gd‐DTPA‐BMA (Omniscan), Gd‐HPDO3A (ProHance), Gd‐DOTA (Dotarem), Gd‐AAZTA, Gd‐BOPTA (Multihance) and Gd‐DTPA (Magnevist)) commonly used in MRI diagnostics. The achieved LODs ranged between 0.40 and 20 μM and the optimized method gave excellent precision, especially when two internal standards were applied (less than 0.34 RSD% for migration time). The MEKC technique made it possible to determine the CAs in urine and serum samples of patients having a therapeutic dose. Due to the SDS content of the running buffer, the serum samples can be directly injected to analyze Gd‐based CAs without interference of high protein content.

A comparative study of capillary electrophoresis and isothermal titration calorimetry for the determination of binding constant of human serum albumin to monoclonal antibody

This paper focuses on the investigation of the interactions between the anti‐HSA‐mAb and its protein antigen using CZE, ACE, and isothermal titration calorimetry. The CZE revealed the formation of the anti‐HSA‐mAb·HSA and anti‐HSA‐mAb·(HSA)2 complexes and the binding constants determined by plotting the amount of the bound anti‐HSA‐mAb as a function of the concentration of HSA. The ACE provided information on the binding strength from the change in effective electrophoretic mobility of the anti‐HSA‐mAb. These two separation techniques estimated the presence of two binding sites. The equilibrium dissociation constant values obtained by CZE and ACE were found to be 2.26 × 10−6 M for anti‐HSA‐mAb·HSA, 1.22 × 10−6 M for anti‐HSA‐mAb·(HSA)2 and 4.45 × 10−8 M for anti‐HSA‐mAb·HSA, 1.08 × 10−7 M for anti‐HSA‐mAb·(HSA)2, respectively. The dissociation constant data obtained by ACE were in congruence with the values obtained by isothermal titration calorimetry (2.74 × 10−8 M, 1.04 × 10−7 M).

Chemometric Assessment of Chromatographic Methods for Herbal Medicines Authentication and Fingerprinting

Nowadays, increasingly more individuals turn to supplementation of the diet with herbal medicines and many such products are marketed lately. Thus the problem that this article focuses on is that these products are not subjected to rigorous quality control like synthetic drugs are, which rises a constant debate whether the supplements actually contain the herb or mixture of herbs that the manufacturer claims they do. As a solution, micellar electrokinetic chromatography and high performance liquid chromatography were investigated in order to fingerprint and authenticate herbal medicines. For this purpose, minimal sample pre-treatment was applied to several fruit based herbal medicines, which were compared with the ethanolic extract of the respective fruit. The holistic evaluation of the electropherograms and chromatograms was made by using appropriate chemometric tools, such as principal component analysis (PCA), cluster analysis and a combination of PCA and linear discriminant analysis (PCA-LDA). The results suggest that the developed method was able to successfully discriminate between different herbal medicines, based on their raw material content. Moreover, this simple and efficient methodology might also be used for routine screening and authenticity control of different products and could be implemented in any quality control laboratory.