Development and validation of a near infrared method for the analytical control of a pharmaceutical preparation in three steps of the manufacturing process

A near infrared diffuse reflectance spectroscopy (NIRS) procedure for the quantitative control analysis of the active compound (otilonium bromide) in a pharmaceutical preparation in three steps of the production process (blended product, cores and coated tablets) and a methodology for its validation are proposed. The analytical procedure is composed by two consecutive steps. First, the sample is identified by comparing its spectrum with a second derivative spectral library. If the sample is positively identified, the active compound is quantified by using a previously established partial least squares (PLS) calibration model. The procedure was validated by studying repeatability, intermediate precision, accuracy and linearity. To this end, an adaptation of ICH (International Conference on Harmonisation) validation methodology to an NIR multivariate calibration procedure is proposed. The relative standard error of prediction (RSEP) was < or = 1% and the suitability of the procedure for control analysis was confirmed by the results obtained analysing new production samples produced over a three-month period.     

Improved ultrasonic extraction procedure for the determination of polycyclic aromatic hydrocarbons in sediments

The aim of this work was to optimize an ultrasonic extraction procedure for the determination of polycyclic aromatic hydrocarbons (PAHs) in sediments and to compare it with the reflux procedure using methanolic potassium hydroxide. Sample extracts were purified with a miniaturized silica gel chromatographic column and analyzed by gas chromatography-mass spectrometry (GC-MS). Ultrasonication using n-hexane-acetone (1:1, v/v) solvent mixture on dried homogenized marine sediment gave better precision (smaller relative standard deviation (RSD) values) and comparable quantities of individual PAH's compared to the reflux procedure. Ultrasonication with the n-hexane-acetone (1:1, v/v) mixture, utilizing four 15 min extraction cycles, was found to be sufficient for extracting PAHs from wet sediments. The optimized ultrasonic extraction procedure extracted aliphatic and aromatic hydrocarbons from the National Institute of Standards and Technology SRM 1941a with recoveries greater than 90%. The major advantages of ultrasonication compared to the reflux method are the lower extraction times, simplicity of the apparatus and extraction procedure. The optimized ultrasonication procedure has been used in our laboratory to extract hydrocarbons from naturally wet sediments from rivers, and coastal and marine areas.     

Development and validation of a near infrared method for the analytical control of a pharmaceutical preparation in three steps of the manufacturing process

A near infrared diffuse reflectance spectroscopy (NIRS) procedure for the quantitative control analysis of the active compound (otilonium bromide) in a pharmaceutical preparation in three steps of the production process (blended product, cores and coated tablets) and a methodology for its validation are proposed. The analytical procedure is composed by two consecutive steps. First, the sample is identified by comparing its spectrum with a second derivative spectral library. If the sample is positively identified, the active compound is quantified by using a previously established partial least squares (PLS) calibration model. The procedure was validated by studying repeatability, intermediate precision, accuracy and linearity. To this end, an adaptation of ICH (International Conference on Harmonisation) validation methodology to an NIR multivariate calibration procedure is proposed. The relative standard error of prediction (RSEP) was ≤ 1% and the suitability of the procedure for control analysis was confirmed by the results obtained analysing new production samples produced over a three-month period.     

流动注射-分光光度-原子吸收串联技术在硅酸盐分析中的应用研究

本文利用流动注射、分光光度、原子吸收联测技术,对硅酸盐系统分析中的Mg、Na、P和Ti进行了Ti-Mg和P-Na联测的技术探讨。装置简单,操作方便,快速。方法用于地质矿产部一级岩石标准参考样,分析结果与推荐值吻合。     

烧结工艺对Ti/IrO2电极在酸性溶液中的电催化活性的影响

采用循环伏安(CV)与极化曲线测试了几种不同烧结工艺制备所得Ti/IrO2电极在酸性Na2SO4溶液中的电催化活性. 对传统单一高温(500 ℃)烧结与改进的分段烧结及程序升温工艺进行了比较; 扫描电镜(SEM)、X射线衍射(XRD)、电化学阻抗谱(EIS)测试表明, 传统工艺所得电极裂纹形貌不明显, 晶粒尺寸及电极的物理电阻均较大, 电化学活性较低; 改进工艺则可明显降低Ti基体的氧化, 提高电极的导电性, 其中程序升温还可使电极表面的裂纹增多, 但若该工艺的起始温度较高, 电极的表观活性下降.     

Ultrasonic energy as a new tool for fast isotopic 18O labeling of proteins for mass spectrometry-based techniques: preliminary results

Preliminary results regarding fast isotopic labeling of proteins with (18)O in conjunction with matrix assisted laser desorption ionization time of flight mass spectrometry technique are presented. Similar (16)O/(18)O isotopic labeling ratios were found for the overnight procedure (12h) and the new fast ultrasonic one (30 min) for the BSA, ovalbumin and alpha-lactalbumin proteins. The procedure, however, failed to promote double (18)O isotopic labeling for the proteins, ovalbumin and alpha-lactalbumin. Two different sonication frequencies, 35 and 130 kHz, were studied at two different sonication times of 15 and 30 min, being best results obtained with the procedure at 130 kHz of sonication frequency and 30 min of sonication time. For comparative purposes the overnight isotopic (18)O labeling procedure was done. In addition, the new fast isotopic labeling procedure was also studied without ultrasonication, in a water bath at 60 degrees C.     

Determination of triphenylphosphine oxide in active pharmaceutical ingredients by hollow-fiber liquid-phase microextraction followed by reversed-phase liquid chromatography

A versatile procedure has been developed and validated for the determination of triphenylphosphine oxide (TPPO) at low levels in various active pharmaceutical ingredients (APIs). This procedure incorporates the use of the novel hollow-fiber liquid-phase microextraction (LPME) for the measurement of this potential process-related impurity in aqueous solutions of APIs. A small volume (40 microL) of 1-octanol contained within a hollow polypropylene fiber is used for the extraction of TPPO from low pH aqueous API solutions. More than a 100-fold increase in the TPPO concentration is obtained without additional evaporation of the extract. Experimental parameters of the extraction procedure were investigated to optimize extraction efficiency and minimize sample matrix interference. Using HPLC/UV as the end analysis technique, the procedure was validated for TPPO in the concentration range of 3-16 microg/L with an API present at 1500 mg/L. The versatility of the method was demonstrated by applying the procedure to the analysis of APIs with different molecular structures. This simple LPME procedure is inexpensive and offers appropriate sensitivity for the intended use while providing several advantages over other analysis methods for pharmaceutical samples.     

One-pot in situ formation and reaction of trimethyl(trichloromethyl)silane: application to the synthesis of 2,2,2-trichloromethylcarbinols

2,2,2-Trichloromethylcarbinols are 1 are valuable synthetic intermediates with a multitude of uses. A scalable procedure for the synthesis of TMS-protected-2,2,2-trichloromethylcarbinols and 2,2,2-trichloromethylcarbinols 1 was developed that employs the in situ generation and reaction of trimethyl(trichloromethyl)silane (CCl(3)-TMS). The procedure avoids the exposure of the carbonyl compounds to the strongly basic conditions typically used for this transformation and also avoids isolation of the difficult-to-handle CCl(3)-TMS. This procedure was applied to diastereoselective trichloromethyl additions to 2-substituted 4-piperidinones and to reactions with a variety of structurally diverse aldehydes and ketones.     

Modification of catalytic adsorptive stripping voltammetric method of hexavalent chromium determination in the presence of DTPA and nitrate

The voltammetric procedure for determination of traces of Cr(VI) [Anal. Chim. Acta (1992) 262:103] was modified by changing the temperature of the measurements. It was found that at the temperature of 40 °C the time of decrease of the Cr(III) signal was shortened from 30 to 5 min. As a result the total analysis time was drastically shortened. The modified procedure does not show any disadvantage as compared to the original method. The results of Cr(VI) determination by the modified procedure are less affected by Cr(III) as compared to the original method. The detection limit of the method was 2.5 × 10^-11 mol L^-1 (1.2 ng L^-1). The validation of the modified procedure was performed by comparison of the results of analyses of tap and river water samples with those obtained using original procedure.     

Development and validation of analytical methodology using capillary electrophoresis for separation and determination of anions in rainwater

A new capillary electrophoresis (CE) procedure was developed for simultaneous determination of both organic and inorganic anions in rain water using a background electrolyte (BGE) containing 5 mM molybdate, 0.15 mM CTAH, 0.01% PVA and 5 mM Tris buffer to adjust pH at 7.9. Under optimised conditions, good repeatability (RSD for sulphate in migration time=0.36% and peak area=4.2%), low detection limit (2 ppb for chloride) and satisfactory working range (50 ppb-20 ppm for hydrodynamic injection, 10 ppb-3 ppm for electrokinetic injection for chloride) were obtained. The reliability of the CE procedure developed was established by satisfactory recovery tests and good agreement of results obtained by both the CE and ion chromatography (IC) methods. The procedure developed had been successfully applied for field monitoring of rainwater showing good repeatability and capability of detecting trace anions at ppb levels beyond the IC working range. Thus, the new CE procedure developed provides a quick, sensitive, economic and reliable method to meet the need for the simultaneous determination of both organic and inorganic anions in the acid rain monitoring programme.     

A New Simple and Sensitive Spectrophotometric Procedure for Determination of Adrenaline

Abstract

A new spectrophotometric procedure is described for the determination of ppm concentrations of adrenaline. The procedure is based on formation of Tris(O-phenanthroline)iron(II) complex (ferroin) upon reaction of adrenaline with an iron(III)-O-phenanthroline mixture in slightly acidic medium. The ferroin complex is then spectrophotometrically measured at 510 nm. In addition to being facile and rapid, the procedure is sufficiently selective and accurate, being particularly suitable for the assay of adrenaline in pharmaceutical formulations; the standard deviation didnot exceed 0.64%.     

Determination of aqueous plutonium oxidation states by solvent extraction

The reliability of two solvent extraction techniques for the determination of Pu oxidation states in solution was tested with low-ionic-strength solutions and with high-Na and high-Mg brines that contained Pu concentrations sufficient for spectrophotometric analysis. One procedure only differentiates between reduced Pu [Pu(III) and Pu(IV)] and oxidized Pu [Pu(V) and Pu(VI)], whereas the second procedure was designed to differentiate between Pu(IV), Pu(V), and Pu(VI) in solution. Both procedures successfully differentiated between oxidized and reduced Pu in both dilute solutions and brines when tested with samples that contained only the Pu(IV), Pu(V), or Pu(VI) oxidation states. However, when the second solvent extraction procedure, which differentiates between Pu(V) and Pu(VI), was employed for solutions that did not contain a strong oxidant to maintain the Pu(VI) oxidation state, significant quantities of Pu(VI) were reduced to Pu(V) during extraction, indicating that accurate quantification of Pu(V) and Pu(VI) is not possible with this procedure.Work supported by the U. S. Department of Energy under Contract DE-ACO6-76RLO 1830.     

High Performance Liquid Chromatographic Determination of (-)-N-Formylnorephedrine in Plasma

Abstract

An HPLC procedure for the detection and quantitative estimation of (-)-N-formylnorephedrine in rabbit plasma had been developed. The procedure involved the extraction of (-)-N-formylnorephedrine from plasma spiked with the internal standard (phenacetin), using ethyl acetate. The ethyl acetate extract is evaporated under nitrogen and the residue is reconstituted in water and injected onto the column. A u-Bondapak-C18 column 30 cm × 3.9 mm ID was used. The mobile phase is 20% acetonitrile in water; at a flow rate of 1.5 ml/min and uv detection at 256 nm. A linear relationship between concentration and peak height ratio (I/internal standard) was obtained (r = 1.00). The reported procedure allows the measurement of (-)-formylnorephedrine in concentrations as low as 150 ng/ml of plasma with total procedure time of about 10 min. The applicability of the procedure to pharmacokinetic studies is illustrated and metabolites are shown not to interfere with the assay procedure.     

Liquid Chromatographic Analysis of Ciprofloxacin and Ciprofloxacin Metabolites in Body Fluids

Abstract

An isocratic HPLC assay procedure for analysis of ciprofloxacin and three metabolites was developed. The procedure requires only dilution of bile, saliva, and urine samples prior to reverse-phase chromatography on a polystyrene-divinylbenzene (PSDVB) column; analysis of serum samples requires a cleanup step on a PSDVB cartridge prior to chromatography. The dependence of chromatographic efficiency on flow rate and temperature was investigated and the accuracy, precision, selectivity, and sensitivity of the procedure were evaluated. The developed procedure was also compared to a modified version of a published ciprofloxacin procedure that requires an octadecyl-silane (ODS) column for chromatographic separation. Similar efficiency, precision, and accuracy were observed with both procedures and both were used for analysis of clinical samples. However, the procedures were used for different purposes. The PSDVB procedure, because of more favorable column selectivity, was used to assay ciprofloxacin and its metabolites in bile, urine and saliva samples. The ODS procedure, because of a simpler serum preparation step, was used t o assay ciprofloxacin in serum samples.     

Supramolecular microextraction of cobalt from water samples before its microsampling flame atomic absorption spectrometric detection

The supramolecular solvent system consists of tetrahydrofuran (THF) and 1-decanol, that was used as an extraction solvent for a microextraction procedure for the preconcentration and separation of Co(II). The proposed supramolecular-based procedure was combined with microsampling flame atomic absorption spectrometry for the determination of cobalt at trace levels in water samples. N-Benzoyl-N,N-diisobutylthiourea was used to chelate Co(II) in an aqueous solution. Quantitative extraction efficiency was obtained at pH 6.5. The effects of analytical parameters including pH, amount of ligand, type, ratio and volume of supramolecular solvent, sample volume and interfering ions were investigated for optimisation of the procedure. The proposed supramolecular solvent-based microextraction procedure (Ss-ME) exhibits a limit of detection (LOD) of 1.29 µg L^?1 and a limit of quantification (LOQ) of 3.88 µg L^?1. The procedure was validated by addition/recovery tests and by applying TMDA 64.2 and TMDA 53.3 water certified reference materials. The microextraction method was successfully applied for the preconcentration and determination of cobalt in water samples.     

Analysis of trace contamination of phthalate esters in ultrapure water using a modified solid-phase extraction procedure and automated thermal desorption-gas chromatography/mass spectrometry

The present study was aimed to develop a procedure modified from the conventional solid-phase extraction (SPE) method for the analysis of trace concentration of phthalate esters in industrial ultrapure water (UPW). The proposed procedure allows UPW sample to be drawn through a sampling tube containing hydrophobic sorbent (Tenax TA) to concentrate the aqueous phthalate esters. The solid trap was then demoisturized by two-stage gas drying before subjecting to thermal desorption and analysis by gas chromatography-mass spectrometry. This process removes the solvent extraction procedure necessary for the conventional SPE method, and permits automation of the analytical procedure for high-volume analyses. Several important parameters, including desorption temperature and duration, packing quantity and demoisturizing procedure, were optimized in this study based on the analytical sensitivity for a standard mixture containing five different phthalate esters. The method detection limits for the five phthalate esters were between 36 ng l(-1) and 95 ng l(-1) and recovery rates between 15% and 101%. Dioctyl phthalate (DOP) was not recovered adequately because the compound was both poorly adsorbed and desorbed on and off Tenax TA sorbents. Furthermore, analyses of material leaching from poly(vinyl chloride) (PVC) tubes as well as the actual water samples showed that di-n-butyl phthalate (DBP) and di(2-ethylhexyl) phthalate (DEHP) were the common contaminants detected from PVC contaminated UPW and the actual UPW, as well as in tap water. The reduction of DEHP in the production processes of actual UPW was clearly observed, however a DEHP concentration of 0.20 microg l(-1) at the point of use was still being quantified, suggesting that the contamination of phthalate esters could present a barrier to the future cleanliness requirement of UPW. The work demonstrated that the proposed modified SPE procedure provided an effective method for rapid analysis and contamination identification in UPW production lines.     

A novel approach for speciation of airborne chromium by convective-interaction media fast-monolithic chromatography with electrothermal atomic-absorption spectrometric detection

A new analytical procedure using an anion-exchange separation support based on convective-interaction media (CIM) was developed for the speciation of chromium. The separation of Cr(VI) was performed on a weak anion-exchange CIM diethylamine (DEAE) fast-monolithic chromatographic disc. Buffer A (0.005 mol dm(-3) TRIS-HCl, pH 8.0) and buffer B (buffer A plus 3 mol dm(-3) NH4NO3) were employed in the separation procedure. The separated chromium species were determined 'off-line' by ETAAS in 0.5 cm3 fractions. The applicability of the CIM DEAE-ETAAS procedure was investigated for the determination of airborne Cr(VI) at a plasma cutting workplace. Aerosols were collected on polycarbonate membrane filters of 8 and 0.4 microm pore size (inhalable and respirable aerosols). Alkaline extraction of filters in a heated ultrasonic bath was applied to leach chromium. Good repeatability of measurement (+/-3.0%) of the alkaline extracts was obtained for Cr(VI). The LOD (3s) was found to be 0.30 microg m(-3) Cr(VI), when 0.25 m3 of air was collected on the filter. The validation of the procedure was performed by spiking filters with Cr(VI) and by the analysis of the standard reference material CRM 545, Cr(VI) in welding dust loaded on a filter. Good recoveries for spiked samples (101-102%) and good agreement between Cr(VI) found and the reported certified value for CRM 545 were obtained. The extracts were also analysed by the FPLC-ETAAS technique. Good agreement between two techniques (r2 = 0.9978) confirmed the reliability of the CIM DEAE-ETAAS procedure developed. The main advantage of the procedure lies in the speed of the chromatographic separation (chromatographic run completed in 15 min).     

Quantitative analysis of morphine in dried blood spots by using morphine-d3 pre-impregnated dried blood spot cards

Two different internal standard dried blood spot (DBS) pre-impregnation procedures (prior to blood spotting) were investigated. In the first procedure DBS pre-impregnation is performed by immersing the DBS card fully into an internal standard solution. In the second procedure pre-impregnation is performed by pipetting a certain volume of an internal standard solution onto the DBS card. Morphine-d3 was used as the model compound for all experiments. The pre-impregnation procedure by immersing was further investigated with respect to homogeneity of impregnation, influence of different blood spotting techniques and the influence of spotting different blood volumes on the internal standard distribution, calibration and stability of pre-impregnated cards. Finally, the immersing procedure was used for the analysis of morphine in dried blood spots and the results were compared to the conventional procedure in which the internal standard morphine-d3 was added to the extraction solvent. The new pre-impregnated cards couple simplicity of operation and convenient use in the field to results equivalent to the conventional procedure.     

The release of heparinase from the periplasmic space of flavobacterium heparinum by three-step osmotic shock

Heparinase was released from the periplasmic space of Flavobacterium heparinum by three-step osmotic shock procedure. The procedure involves resuspending exponentially growing cells consecutively into (1) 40% sucrose, (2) 10 mM sodium phosphate, 2 mM magnesium chloride, pH 7, and (3) 10 mM sodium phosphate, 300 mM sodium chloride, 2 mM magnesium chloride, pH 7. Typically, 50-75% of the total heparinase activity is recovered by this procedure with an observed 7-15-fold increase in purity. The majority of heparinase activity is released in the final step of the procedure allowing for resolution from cytoplasmic and nonspecific periplasmic material. F. heparinum cells can be stored in 40% sucrose at 4 degrees C for up to one week without significant losses in recovery yields.     

高效液相色谱-旋光检测器联用实时检测对映体过量

在手性合成或拆分过程中,实时监测反应产物的对映体过量(e.e.)对于确定产物光学纯度至关重要。根据在一定测定条件下,单一对映体旋光度仅与其浓度成正比,利用高效液相色谱与旋光检测器联用,提出了一种实时测定e.e.的新方法。并在猪胰脂肪酶催化酯交换反应制备(S)-2-甲基-1-丁醇的过程中,比较了该方法与常规测定方法,证实了该方法简单,快速,有望作为一种手性技术中e.e.在线测定通用方法。