Automated, on-line membrane extraction

Over the last few years, membranes have been used to develop new approaches in analytical extraction, concentration and cleanup. An important advantage of membrane processes is that the sample and the extraction phase can be continuously brought into contact without physical mixing, and may be directly interfaced to an analytical instrument. This provides the basis for automated, real-time monitoring. Membrane extraction has been applied to a wide range of organic and inorganic analytes, and has been directly interfaced with chromatography, spectroscopy and mass spectrometry. Implementations of membrane extraction are diverse, encompassing different types of membranes, module designs and configurations. This review highlights some of these, and particularly the unique capabilities in automated, and on-line measurements.     

Normal-phase high-performance liquid chromatographic determination of phenols

Normal-phase high-performance liquid chromatography has been used for separation of phenol and its monoderivatives. Multi-component mixtures of hexane (non-polar component) with butan-1-ol, chloroform, butyl bromide, butyl chloride or diethyl ether (polar additives) were used as selective eluents. Silica gel "Silasorb 600" with specific surface area of about 600 m(2)/g and average particle size of $ 10 mum was used as the sorbent. Phenol and the o-, m-, p-isomers of cresol were concentrated by extraction with n-butyl acetate from aqueous solutions. A method for determination of microamounts of phenols in aqueous solutions in the presence of 160-fold amounts of aromatic hydrocarbons has been developed.     

Automated high-performance liquid chromatographic determination of chloramphenicol in milk and swine muscle tissue using on-line immunoaffinity sample clean-up.

An on-line high-performance liquid immunoaffinity chromatographic (HPLIAC) system for the direct determination of chloramphenicol in milk and swine muscle tissue is described. The system consisted of a dual-column system in which an HPLIAC column was directly coupled to an RP-8 high-performance liquid chromatographic column. Skimmed and deproteinated milk or aqueous muscle tissue extract was directly injected into the HPLIAC column. After a washing step with phosphate-buffered saline, chloramphenicol was desorbed by a glycine-NaCl buffer (pH 2.8) and directly concentrated on the RP-8 column. Next, chromatography was carried out using acetonitrile-sodium acetate buffer as the mobile phase. Chloramphenicol was detected at 280 nm. Mean recoveries from spiked raw milk were 70 +/- 2% (1-50 micrograms/kg) and from spiked swine muscle tissue 64 +/- 2% (10-50 micrograms/kg). The calibration curves were linear in the range 1-200 micrograms/kg spiking levels. Limits of determination were 1 microgram/kg for milk and 10 micrograms/kg for muscle tissue.     

Evaluation of an on-line coupled continuous flow liquid membrane extraction and precolumn system as trace enrichment technique by liquid chromatographic determination of bisphenol A

An on-line coupled continuous flow liquid membrane extraction (CFLME) and C₁₈ precolumn system was developed for sample preconcentration in liquid chromatography determination. After preconcentration by CFLME, which is based on the combination of continuous flow liquid–liquid extraction and supported liquid membrane, bisphenol A (BPA) was enriched in 960 μl of 1 mol l⁻¹ NaOH used as acceptor. This acceptor was on-line neutralized and transported onto the C₁₈ precolumn where analytes were absorbed and focused. Then the focused analytes were injected onto a C₁₈ analytical column for separation and detected at 220 nm with a diode array detector. CFLME related parameters such as flow rates, pH of donor and acceptor, and enrichment time were optimized. The proposed method presents a detection limit of 0.03 μg l⁻¹ (S/N=3) when 60 ml samples was enriched with an enrichment time of 30 min. Compared with C₁₈ based column-switching procedure, this proposed procedure presents similar sample throughput and lower detection limits. The proposed method was successfully applied to determine BPA in tap water, river water, and municipal sewage effluent samples.     

High-performance liquid chromatographic determination of phenolic compounds in natural water coupled with on-line flow injection membrane extraction-preconcentration

A liquid chromatographic method for determination of trace phenolic compounds has been established, coupled with an on-line supported liquid membrane extraction-preconcentration flow-injection system. Tributyl-phosphate dissolved in kerosene was used as the carrier of the supported liquid membrane. Four phenolic compounds (phenol, catechol, resorcinol and hydroquinone) were chosen as the model compounds and the experiment conditions were optimized. Under the optimum conditions, calibrations were linear in the range of 1-500 microg/L, with good correlation coefficients (r > 0.999). The total analysis time of the system was 22 min, including the membrane extraction, liquid chromatographic separation and equilibration times.     

Photometric determination of phenols in essential oils

A photometric procedure is described for determining terpene phenols in essential oils which is based on the formation of colored indophenols with p-quinone chloroimine in an alkaline medium. A comparative evaluation of the results obtained with those of the pharmacopoeial method shows the greater economy, sensitivity, and accuracy of the proposed procedure. Zaporozh'e Medical Institute. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 509–511, July–August, 1988.     

Automated high-performance liquid chromatographic assay for the determination of 7-ethoxycoumarin and umbelliferone.

An improved high-performance liquid chromatographic assay is presented for the determination of 7-ethoxycoumarin O-deethylase activity. Following a 30-min microsomal incubation, 7-ethoxycoumarin, 4-methylumbelliferone (internal standard), and the metabolite umbelliferone were extracted with chloroform. Separation was achieved with an isocratic mobile phase using a microBondapak phenyl (300 mm x 3.9 mm I.D.) analytical column. The effluent was monitored by fluorescence detection with an excitation wavelength of 360 nm and an emission wavelength of 470 nm. The intra- and inter-assay coefficients of variation were 10 and 6%, respectively. A detection limit of 0.07 micrograms/ml was achieved, making this method suitable for characterizing P-450 activity of human livers.     

Photometric determination of phenols in essential oils

A photometric procedure is described for determining terpene phenols in essential oils which is based on the formation of colored indophenols with p-quinone chloroimine in an alkaline medium. A comparative evaluation of the results obtained with those of the pharmacopoeial method shows the greater economy, sensitivity, and accuracy of the proposed procedure.Zaporozh'e Medical Institute. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 509–511, July–August, 1988.     

High-performance liquid chromatography-fluorescence determination of dinitropyrenes in soil after column chromatographic clean-up and on-line reduction.

In order to quantify 1,3-dinitropyrene (DNP), 1,6-DNP and 1,8-DNP in soil, we developed an efficient clean-up procedure and a sensitive determination method using fluorescence detection. DNP isomers were efficiently cleaned by three stages of fractionation, i.e., a silica gel open column chromatography using stepwise elution and two further purification steps by high-performance liquid chromatography (PPLC) using a monomeric-type octadecylsilyl (ODS) column and a polymeric-type ODS column. The recoveries of DNPs during the whole clean-up process were 94% or more. The fraction corresponding to DNPs was injected into an analytical polymeric-type ODS column for HPLC to separate DNP isomers. The effluent from the analytical ODS column was directly introduced to a catalyst column, which was packed with 5 microns alumina coated with platinum and rhodium (Pt-Rh), in order to reduce DNPs to diamino compounds, and then the fluorescence of diaminopyrenes was detected. The immediate detection of diaminopyrene isomers after on-line reduction afforded a sensitive detection of DNP isomers. The detection limits for DNPs were in the range of 0.7 to 4 pg. These developed methods were applied to four soil samples collected at parks in residential areas.     

Determination of sterols, erythrodiol, uvaol and alkanols in olive oils using combined solid-phase extraction, high-performance liquid chromatographic and high-resolution gas chromatographic techniques.

A method is described for the determination of the sterol, erythrodiol, uvaol and alkanol content in olive oils by means of solid-phase extraction and high-performance liquid chromatography, instead of liquid-liquid and thin-layer chromatographic separations, the following step being high-resolution gas chromatographic separation. This type of procedure allows the simultaneous analysis of a larger number of samples and a substantial reduction in manual operations. Comparisons were made between the two methods on 100 different olive oils and with a statistical analysis of the results (Student's t-test).     

Automated liquid chromatographic determination of ranitidine in microliter samples of rat plasma

An improved high-performance liquid chromatographic method with UV detection at 313 nm has been developed for quantitation of ranitidine in 100 microliter of rat plasma over the range 25 to 1000 ng/ml. To each sample were added the internal standard (metiamide) and 2 M NaOH. After dichloromethane extraction, the nitrogen-dried extracts were reconstituted in the mobile phase of 0.01 M phosphate buffer-triethylamine-methanol-water (530:5:390:75 v/v). Chromatography on mu Bondapak C18 with quantitation by peak height ratios showed an analyte recovery of 97%; a limit of detection of 10 ng/ml; a precision of 1-10% and an accuracy of 1-5%. About 90 samples can be processed in 24 h.     

Automated high-performance liquid chromatographic determination of 5-S-cysteinyl-3,4-dihydroxyphenylalanine in urine

An automated high-performance liquid chromatographic (HPLC) method has been developed for measurement of 5-S-cysteinyl-DOPA in urine (DOPA = 3,4-dihydroxyphenylalanine). The urinary sample was injected into an HPLC boronate column. With a mobile phase of 0.1 M phosphate buffer containing 0.2 mM disodium ethylenediaminetetraacetate (Na2EDTA) (pH 6.0) mixed with methanol (9:1), 5-S-cysteinyl-DOPA was adsorbed while most other compounds were washed away. By column switching, the column flow was reversed and 5-S-cysteinyl-DOPA was desorbed by a mobile phase of 0.1 M formic acid and 0.2 mM Na2EDTA at pH 3.0 and chromatographed on a reversed-phase column. The precision, as estimated from repeated analysis of an urinary sample and from duplicate analysis of a number of samples, ranged from 1.4 to 5.2% (coefficient of variation), and the analytical recovery was 93 +/- 4.1%. The method is suitable for use in the clinical laboratory.     

Automated on-line multi-dimensional high-performance liquid chromatographic techniques for the clean-up and analysis of water-soluble samples

The application of an automated on-line multi-dimensional liquid-liquid chromatographic technique for the clean-up and analysis of water-soluble samples was investigated. The use of microparticulate aqueous-compatible steric exclusion columns as the primary separation step coupled to either reversed-phase, normal-phase or ion-exchange columns as the secondary step allowed the direct injection of complex samples without prior clean-up. The entire operation was automatically controlled by a microprocessor-based liquid chromatograph with time-programmable events which allowed precise switching of high-pressure pneumatically operated valves. Both heart-cutting and on-column concentration methods were used. The heart-cutting technique had the advantage of selectivity but lacked sensitivity; more successful was the on-column concentration technique, which, by the concentration of the solute from a larger volume of exclusion column effluent on to the secondary column, gave better sensitivity. The technique was applied to the analysis of theophylline and caffeine in biological fluids, catecholamines in urine, vitamins in a protein food supplement and sugars in molasses and candy bars.     

Automated liquid chromatographic analysis of drugs in urine by on-line sample cleanup and isocratic multi-column separation

A multi-column system has been developed for automated analysis of basic drugs in urine. Two polymeric pre-columns, containing PRP-1 and Aminex A-28, were used to isolate the drugs. A short reversed-phase column, coupled to a 150 x 4.6 mm I.D. silica column, produced the analytical separation. Sample preparation consisted of dilution and centrifugation. The entire procedure required less than 30 min. Careful optimization of mobile phase conditions led to retention of benzoylecgonine and barbiturates. For most drugs, levels of 0.3 mg/l were sufficient to produce peaks that could be matched against stored spectra with a computerized library search program.     

Multidimensional gas chromatographic determination of naphthenes, paraffins, olefins, and aromatics in reformed gasolines

Increasing demand for gasoline, changing regulations concerning the reduction of environmental impact, and new refining technologies have led to the refinement of its composition. Nowadays, gasoline is a complex mixture of different fractions deriving from processes of reforming, cracking, isomerization, and alkylation, with the addition of both oxygenated compounds and butanes. There are regulations governing the mixing of various fractions and it is necessary to analyse the composition of these fractions to ensure that the final composition of commercial gasoline satisfies the required specifications. Moreover, analysis of the composition of each fraction enables the technological process of the fraction examined to be modified as appropriate. In this work some reformed gasolines were analysed by multidimensional gas chromatography. This technique allows good separation of the hydrocarbon types in a single analysis and gives the carbon number distribution within each hydrocarbon type.     

Determination of phenols in water by on-line solid-phase disk extraction and liquid chromatography with electrochemical detection

Poly(styrene-divinylbenzene) (PS-DVB) membrane extraction disks were used as sorbents for the on-line solid phase extraction of 13 phenols (nitro and chlorophenols) in river and tap waters. Determination was performed by liquid chromatography with electrochemical detection (LC-ED). An acetate buffer-acetonitrile-methanol mixture as mobile phase and amperometric detection at +1100 mV were used. High water volumes, up to 250 ml, can be preconcentrated without loss of phenols (recoveries between 80% and 100%) except for the more polar ones. Moreover, detection limits between 0.01 and 0.1 μg l⁻¹ in tap water and between 0.1 and 1.0 μg⁻¹ in river water were obtained. The method has been applied to the analysis of two river water samples.