Effect of a nano-sized natural clinoptilolite modified by the hexadecyltrimethyl ammonium surfactant on cephalexin drug delivery

The use of porous materials as host systems for medical applications has been considered in recent years. The aim of this work is to construct an efficient adsorbent for the adsorption and delivering of cephalexin. For this pupose, pretreated natural nano-sized clinoptilolite (NZ) was modified by the cationic hexadecyltrimethyl ammonium surfactant (HDTMA), and the obtained modified zeolite nanoparticles (SMZ) were used to design systems for storage and release of cephalexin (CPX). The adsorbed and released extents of the drug onto/from the modified zeolite were determined by UV–Vis spectroscopy. The results showed that both decreasing the particle size of clinoptilolite and modifying its surface significantly increase the adsorbed drug extent. All the compounds were characterized by SEM, TEM, FT–IR, TG/DTG, and XRD. TG/DTG and also FT–IR results showed sufficient loading amounts of HDTMA and CPX onto the raw and modified zeolite, respectively. It was proven by means of TG that the composites are more stable thermally when the admicelles contain cephalexin in their interior. IR spectroscopy studies indicated that the zeolite structure remained unchanged after the modification with the surfactant and after the cephalexin drug has been loaded. Due to the presence of hydroxyl and amine groups in the cephalexin structure, pH plays an important role on the adsorbed CPX extent, so that the maximum adsorbed CPX was observed at pH = 12. While the delivery of CPX was better at pH = 2, because at alkaline pHs, the anionic carboxylate form of CPX has higher attractive force with the positive head of the surfactant on the SMZ. Hence, the stomach's acidic pH is appropriate for drug delivery. The effects of some cations in the delivery extent confirm that the diet can significantly affect the delivery of the CPX from the proposed adsorbent.     

Development of enzyme-linked immunosorbent assays for the detection of deacetoxycephalosporin C and isopenicillin N synthase activity

Although there are a number of existing assays for monitoring the activity of both isopenicillin N synthase (IPNS) and deacetoxycephalosporin C synthase (DAOCS), none have demonstrated the qualities required for screening a mutant library. Hence, enzyme-linked immunosorbent assays (ELISAs) for IPNS and DAOCS were developed based on the detection of the catalytic turnover products isopenicillin N and cephalexin/phenylacetyl-7-aminodeacetoxycephalosporanic acid (G-7-ADCA), respectively. These assays are relatively fast compared to existing assays, such as the hole-plate bioassay, and are amenable with high-throughput screening. Both the IPNS and DAOCS-ELISAs were optimised for use with crude protein extracts rather than purified protein, thereby eliminating any additional time required for purification. The ELISA developed for the detection of cephalexin had an IC₅₀ value of 154 ± 9 ng mL⁻¹ and LOD of 7.2 ± 2.2 ng mL⁻¹ under conditions required for the assay. Good recoveries and correlation was observed for spiked samples when the concentration of crude protein was kept below 1 mg mL⁻¹. The DAOCS-ELISA was found to have increased sensitivity compared to the hole-plate bioassay (10.3 μg mL⁻¹). The IPNS-ELISA did not significantly increase the sensitivity (approximately 5 μg mL⁻¹) compared to that of the hole-plate bioassay (16 μg mL⁻¹) for isopenicillin N. The minimum amount of crude protein extract required for producing detectable amounts of product for both assays was below 0.5% of the maximum amount of protein that the assay could contain without any effect on the ELISA. This suggests that when screening a mutant library, mutants producing low amounts of the product could still be detected using these assays.     

Spectrofluorometric Determination of Cephalexin in Pharmaceutical Preparations and Spiked Human Urine Using 2‐Cyanoacetamide

A simple, accurate, sensitive, and validated method was developed for the spectrofluorometric determination of cephalexin. The method involves the reaction of cephalexin with 2‐cyanoacetamide in presence of 33% ammonia solution. The formed fluorescent product exhibited maximum fluorescence intensity at λ 439 nm, after excitation at λ 339 nm. Different experimental parameters affecting the derivatization reaction were carefully studied and incorporated in the procedure. Under the described conditions, the proposed method was linear over the concentration range 0.04–0.4 µg/mL. The average percent found was 99.6±0.9%. The LOD was 7.76 ng/mL. The proposed method was applied for determination of cephalexin in pharmaceutical preparations as well as in spiked human urine. A mechanism of the reaction is postulated.     

Spectrophotometric Determination of Certain Cephalosporins Using Ferrihydroxamate Method

Cephalexin, cefixime, ceftriaxone and cefotaxime were determined spectrophotometrically in the pure form and in pharmaceutical formulations by using ferrihydroxamate method. Reaction optimization with respect to reaction time and temperature has been investigated. Influence of the presence of ester functional group on the determination of cephalosporins as ß-lactams under conditions optimized was evaluated. Using cefotaxime sodiume as model drug with ester functional group, it was shown that proposed method gives equally acurate and precise results even in the presence of ester functional group.     

大网格吸附剂提取头孢氨苄

本文研究了用大网格吸附剂提取头孢氨苄的方法，包括吸附剂和解吸剂的筛选，吸附条件和解吸条件的考察。确定了用H—103型树脂在pH5、流速1／10（V／V／min）条件下吸附；用50％丙酮在pH1．5、流速1／30（V／V／min）条件下解吸。吸附容量可达75mg／ml，是国内报道过的CAD—40的3倍，是国外报道过的DiaionHP－20的3．8倍，解吸率可达92％以上。     

Development of a new high performance low pressure chromatographic system using a multisyringe burette coupled to a chromatographic monolithic column

A novel combination of high performance low pressure chromatography with multisyringe flow injection analysis is presented. This system comprises a multisyringe module, three low pressure solenoid valves, a monolithic Chromolith Flash RP-18e column and a diode array spectrophotometer. UV detection is carried out at 250 nm. AutoAnalysis software is used for instrumental control and automated data collection. The results obtained with multisyringe liquid chromatography (MSC) were compared with those obtained with a HPLC system using similar conditions. The chromatographic parameters were calculated from a mixture of anthracene and thiourea using a mobile phase containing acetonitrile-water (60:40) at a flow rate of 2 ml min⁻¹. The proposed MSC system has been successfully applied to the determination of amoxicillin, ampicillin and cephalexin using a mobile phase of sodium acetate buffer (pH 6.2, 0.1 mol l⁻¹)-methanol (90:10) at a flow rate of 2 ml min⁻¹. The low-cost, flexibility and simplicity of MSC should be highlighted.     

银微盘电极上头孢氨苄降解产物的伏安行为及其应用

基于头孢氨苄降解产物中含有巯基，能与Ａｇ＾＋生成难溶化合物的特性，在银微盘电极上研究了ＣＥＸ降解产物的伏安行为，探讨了电极反应的机理。运用示差脉冲溶出伏安法，在０．１ｍｏｌ／Ｌ ＨＡｃ－ＮａＡｃ介质中，ＣＥＸ降解产物的还原峰电流与ＣＥＸ浓度于８．０×１０＾－８－７．０×１０＾－６ｍｏｌ／Ｌ范围内呈良好线性关系；检测限为１．０×１０＾０８ｍｏｌ／Ｌ；测定了头孢氨苄胶囊中ＣＥＸ含量为９２．０％，与药典     

光化学荧光分析法测定头孢氨苄的研究

在ｐＨ４．０的缓冲溶液中，头孢氨苄在紫外光照射下能发生光化学反应，形成具有荧光的产物，λｅｘ为３４５ｎｍ，λｅｍ为４３２ｎｍ。在头孢氨苄浓度为０．１－４．０μｇ／ｍｌ范围内，荧光强度与浓度呈良好的线性关系，由此建立了头孢氨苄的光化学荧光测定法。方法的检出限为０．０１μｇ／ｍｌ，相对标准偏差为０．９３％。本法简便，快速且灵敏度高，可用于尿液中头孢氨苄含量的测定。     

A new derivatization procedure for the determination of cephalexin with 1,2-naphthoquinone 4-sulphonate in pharmaceutical and urine samples using solid-phase extraction cartridges and UV–visible detection

The present report shows how to derivatize cephalexin with 1,2-naphthoquinone-4-sulphonate (NQS) into solid-phase extraction cartridges (C₁₈) using UV–visible detection. Optimum conditions for this new procedure are: hydrogen carbonate/carbonate buffer pH=10.5, 5 min reaction time at 25°C and NQS concentration of 7.1×10⁻³ mol l⁻¹. The accuracy and the precision of the method were tested. The procedure was used to measure cephalexin in pharmaceutical and urine samples. The results obtained were contrasted with those reported by UV-spectrophotometric and HPLC methods for pharmaceutical samples and with HPLC method for urine samples. The H-point standard additions method was used to measure cephalexin in pharmaceutical samples, and the generalized H-point standard additions method was used to measure cephalexin in urine samples.     

Molecularly imprinted solid phase extraction for rapid screening of cephalexin in human plasma and serum

A molecularly imprinted solid phase extraction (MISPE) method was developed for the rapid screening of cephalexin in human plasma and serum. The method employed a micro-column packed with molecularly imprinted polymer (MIP) particles for the selective solid phase extraction (SPE) of cephalexin. Since the MIP interacted indiscriminately with two other α-aminocephalosporins, cefradine and cefadroxil, their removal was ultimately achieved using differential pulsed elution (DPE) with acetonitrile+12% acetic acid. Cephalexin was then determined in a final pulsed elution (FPE) with methanol+1% trifluoroacetic (CF₃COOH, TFA) acid. This excellent selectivity represents a significant advance in analytical separation, demonstrating how a MIP can differentiate between molecules that are structurally dissimilar only in their non-hydrogen-bonding moieties, even if their hydrogen-bonding moieties are identical to each other. With UV detection, a concentration detection limit of 0.1 μg/ml (or 2 ng in 20 μl) was afforded for cephalexin. By increasing the CHCl₃ flow rate to 1.25 ml/min, each MISPE-DPE-FPE analysis required only 2 min to complete. Rapid screening was demonstrated in a modified MISPE-PE method, which used 14% CH₃COOH+CH₃CN as the mobile phase, followed by direct PE with 1% TFA+CH₃OH.