Liquid Chromatographic System For Separation Research Articles

Development of an analytic method for organosulfur compounds in Welsh onion and its use for nutritional quality analysis of five typical varieties in China

A reliable, simple, and sensitive method capable of quantifying six organosulfur compounds (OSCs) was established. The samples were extracted by water containing 3 % formic acid with a simple vortex, ultrasound, and centrifugation step, and the solutions were analyzed by ultra-high-performance liquid chromatography separation system coupled with a triple-quadrupole mass spectrometry (UHPLC − MS/MS). Then the method was applied for the analysis of six OSCs in five varieties of two types Welsh onions in China, and the moisture content, reducing sugar, total polyphenols, and 21 free amino acids were also analyzed to study the characters of these Welsh onions intensively. Multivariate statistical analysis was used to investigate the differences in OSCs and free amino acids profiles among the samples. This study showed that enzymatic inhibition method combined with UHPLC − MS/MS is an effective technique to analyze OSCs in Welsh onion, and could be valuable for the routine quantitation of OSCs in other foods.

Rapid and sensitive dilute-and-shoot analysis using LC-MS-MS for identification of multi-class psychoactive substances in human urine

The emergence of new psychoactive substances currently exceeding a thousand is rapidly changing substance prevalence patterns and straining the methods used for detection, most of which are suitable only for a single class of substances. This study presents a rapid and facile dilute-and-shoot system operated in conjunction with an optimized liquid chromatographic separation system for the high-sensitivity detection of substances across a range of substance classes with 3 isotopes used only. The proposed method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) is able to identify 68 substance and their metabolites in urine samples as small as 50 μL. Optimal chromatographic conditions including 95% water/methanol ratio with 0.1% added formic acid and a prolonged LC gradient run-time (15 min) improved the peak shape of polar compounds and enhanced signal strength by 5%. Under 4-fold dilution, all analytes were within 80–120% of tolerance response levels, indicating that the matrix effect was insignificant. In experiments, the limit of detection (LOD) ranged from 0.05 to 0.5 ng mL−1, while the coefficient of determination (R2) was > 0.9950. The retention time shift of each peak remained at < 2% with an inter-day relative standard deviation (RSD) of 0.9–14.9% and intra-day RSD of 1.1%− 13.8%. The rapid dilute-and-shoot presents a high-sensitivity, significant stability, robustness and reproducibility without serious interference. To demonstrate the effectiveness of the system, 532 urine samples were collected from suspected drug abusers, and the proposed method was used for rapid analysis. Of these samples, 79.5% contained between one and twelve analytes, and 12.4% tested positive for new psychoactive substances, mostly derivatives of amphetamine and synthetic cathinones. The study presents a high-sensitivity analytic system that is capable of detecting substances from multiple classes and can be used for effective monitoring of substance prevalence in urine.

A Streamlined High-Throughput Plasma Proteomics Platform for Clinical Proteomics with Improved Proteome Coverage, Reproducibility, and Robustness.

Mass spectrometry-based clinical proteomics requires high throughput, reproducibility, robustness, and comprehensive coverage to serve the needs of clinical diagnosis, prognosis, and personalized medicine. Oftentimes these requirements are contradictory to each other. We report the development of a streamlined High-Throughput Plasma Proteomics (sHTPP) platform for untargeted profiling of the blood plasma proteome, which includes 96-well plates and simplified procedures for sample preparation, disposable trap column for peptide loading, robust liquid chromatographic system for separation, data-independent acquisition in tandem mass spectrometry, and DIA-NN, FragPipe, and in-house peptide spectral library-based data analysis. Using the optimized platform at a throughput of 60 samples per day, over 600 protein groups including 57 FDA-approved biomarkers can be consistently identified from whole human plasma, and more than 85% of the detected proteins have 100% completeness in quantitative values across 300 samples. The balance achieved between proteome coverage, throughput, and reproducibility of this sHTPP platform makes it promising in clinical settings, where a large number of samples are to be measured quickly and reliably to support various needs of clinical medicine.

Cellulose tris-(3,5-dimethylphenylcarbamate)-based chiral stationary phase for the enantioseparation of drugs in supercritical fluid chromatography: comparison with HPLC.

A cellulose tris-(3,5-dimethylphenylcarbamate)-based chiral stationary phase was studied as a tool for the enantioselective separation of 21 selected analytes with different pharmaceutical and physicochemical properties. The enantioseparations were performed using supercritical fluid chromatography. The effect of the mobile phase composition was studied. Four different additives (diethylamine, triethylamine, isopropylamine, and trifluoroacetic acid) and isopropylamine combined with trifluoroacetic acid were tested and their influence on enantioseparation was compared. The influence of two different mobile phase co-solvents (methanol and propan-2-ol) combined with all the additives was also evaluated. The best mobile phase compositions for the separation of the majority of enantiomers were CO2 /methanol/isopropylamine 80:20:0.1 v/v/v or CO2 /propan-2-ol/isopropylamine/trifluoroacetic acid 80:20:0.05:0.05 v/v/v/v. The best results were obtained from the group of basic β-blockers. A high-performance liquid chromatography separation system composed of the same stationary phase and mobile phase of similar properties prepared as a mixture of hexane/propan-2-ol/additive 80:20:0.1 v/v/v was considered for comparison. Supercritical fluid chromatography was found to yield better results, i.e. better enantioresolution for shorter analysis times than high-performance liquid chromatography. However, examples of enantiomers better resolved under the optimized conditions in high-performance liquid chromatography were also found.

Two-dimensional liquid chromatography separation and high resolution mass spectrometry analysis for proteome of rice leaves based on different extraction methods

A two-dimensional liquid chromatography (2D LC) system was developed to separate proteins from rice leaves, which was extracted by phenol method, followed by the analysis with linear trap quadrupole orbitrap mass spectrometry (LTQ/Orbitrap MS). After proteins were extracted with phenol method, the enzymolytic peptides were separated by offline two-dimensional RP-RP system and detected by LTQ/Orbitrap MS, yielding 2712 proteins. Liquid chromatography separation system (1D LC and 2D LC) and protein extraction methods (phenol method, sodium dodecyl sulfate method (SDS method) and trichloroacetic acid/acetone method (TCA/acetone method)) were compared. Proteins identified by 2D LC were 2712, 2415 and 1914 with the above three extraction methods, respectively. The proteins were 2.7-fold, 2.5-fold and 1.9-fold the number of proteins identified by 1D LC respectively. And in terms of 2D LC, the proteins identified by phenol method were 297 and 798 more than SDS method and TCA/acetone method, respectively. Some proteins with extreme properties, such as very acidic or basic protein and high relative molecular mass proteins, were only identified in phenol method. Furthermore, proteins, which were extracted by different extraction methods and separated by 2D LC, were classified according to biological functions. It was found that protein functions by the three extraction methods were complementary. However, phenol method had the most variety of functions. The method provides technological support for rice proteomics and reference for research techniques of other crop proteomics.

Simultaneous screening for lipophilic and hydrophilic toxins in marine harmful algae using a serially coupled reversed-phase and hydrophilic interaction liquid chromatography separation system with high-resolution mass spectrometry

The presence of toxins in harmful algal blooms (HABs) poses considerable concerns because of their potential adverse effects on ecological environments and human health. When marine HABs occur, efficient screening and identification of toxins in different kinds of HAB algae remains a challenge. In this study, the applicability of serial coupling of reversed-phase liquid chromatography (RPLC) and hydrophilic interaction chromatography (HILIC) combined with high resolution mass spectrometry (HR-MS) for the simultaneous screening and identification of various kinds of known lipophilic and hydrophilic toxins in HAB algae was investigated for the first time. Ultrasound-assisted extraction (UAE) was explored to extract both lipophilic and hydrophilic toxins in algae simultaneously. As in most cases, toxin standards were not available; therefore, an identification procedure based on accurate mass data and chromatographic behavior was proposed. According to this procedure, eight known lipophilic toxins and 11 hydrophilic toxins were successfully detected in a single injection, and the proposed method was validated. Satisfactory sensitivity, repeatability (RSD <14.87%) and recovery (89.4–105.8%) of the method were achieved. A major advantage of the proposed method is that it can almost detect members of all eight groups of marine algal toxins in a single run. Using this method, several known toxins in different marine toxigenic algae including Alexandrium tamarense, Alexandrium minutum and Prorocentrum lima were successfully observed and identified. This work demonstrates that RPLC/HILIC-HR-MS combined with an accurate mass list of known marine algal toxins may be used as a powerful tool for screening of different classes of known toxins in marine harmful algae.

Simultaneous Online Enrichment and Identification of Trace Species Based on Microfluidic Droplets

A facile online enrichment protocol has been proposed based on microfluidic droplets acting as an interface between a liquid chromatography separation system and detection systems of ESI-MS/MS and laser-induced fluorescence. Low-abundance species were successfully concentrated and analyzed in this system via droplet shrinkage. The proposed platform significantly increased the enrichment efficiency and detection sensitivity with reduced sample handling steps, short analysis time, and no cross-contamination. The presented system is universal, shows no discrimination, and is easily coupled with other separation and detection approaches.

Comparative analysis of nucleosides and nucleobases from different sections of Elaphuri Davidiani Cornu and Cervi Cornu by UHPLC–MS/MS

A rapid and sensitive hydrophilic interaction liquid chromatography method was developed for the identification and quantitative determination of 17 nucleosides and nucleobases by using ultra-high performance liquid chromatography separation system coupled to triple quadrupole mass spectrometer. Sufficient separation of the target compounds was achieved on an Acquity BEH Amide column (2.1mm×100mm, 1.7μm) in a single run of 9min. The LOD and LOQ were in the range of 0.017–50.00ngml−1 and 0.056–146.00ngml−1, respectively. Then, the method was applied for the analysis of nucleosides and nucleobases in Elaphuri Davidiani Cornu and Cervi Cornu. Multivariate statistical analysis was also used to investigate the differences in the nucleosides and nucleobases profiles among the samples. The present study also provided helpful information for quality control of Elaphuri Davidiani Cornu and Cervi Cornu.

Comparison of optimised germanium gamma spectrometry and multicollector inductively coupled plasma mass spectrometry for the determination of 134Cs, 137Cs and 154Eu single ratios in highly burnt UO2

Non-destructive and destructive methods have been compared to validate their corresponding assessed accuracies in the measurement of 134Cs/137Cs and 154Eu/137Cs isotopic concentration ratios in four spent UO2 fuel samples with very high (52 and 71GWd/t) and ultra-high (91 and 126GWd/t) burnup values, and about 10 (in the first three samples) and 4 years (in the latter sample) cooling time. The non-destructive technique tested was high-resolution gamma spectrometry using a high-purity germanium detector (HPGe) and a special tomographic station for the handling of highly radioactive 400mm spent fuel segments that included a tungsten collimator, lead filter (to enhance the signal to Compton background ratio and reduce the dead time) and paraffin wax (to reduce neutron damage). The non-destructive determination of these isotopic concentration ratios has been particularly challenging for these segments because of the need to properly derive non-Gaussian gamma-peak areas and subtract the background from perturbing capture gammas produced by the intrinsic high-intensity neutron emissions from the spent fuel. Additionally, the activity distribution within each pin was determined tomographically to correct appropriately for self-attenuation and geometrical effects. The ratios obtained non-destructively showed a 1σ statistical error in the range 1.9–2.9%. The destructive technique used was a high-performance liquid chromatographic separation system, combined online to a multicollector inductively coupled plasma mass spectrometer (HPLC-MC-ICP-MS), for the analysis of dissolved fuel solutions. During the mass spectrometric analyses, special care was taken in the optimisation of the chromatographic separation for Eu and the interfering element Gd, as also in the mathematical correction of the 154Gd background from the 154Eu signal. The ratios obtained destructively are considerably more precise (1σ statistical error in the range 0.4–0.8% for most of the samples, but up to 2.8% for one sample). The HPGe gamma spectrometry can achieve a high degree of accuracy (agreement with HPLC-MC-ICP-MS within a few percent), only by virtue of the optimised setup, and the refined measurement strategy and data treatment employed.

LC‐MS/MS in the elucidation of an isomer of the recreational drug methylenedioxy ethylamphetamine: Methylenedioxy dimethylamphetamine

This paper describes the surplus value of a quadrupole-orthogonal acceleration TOF mass spectrometer, coupled to a liquid chromatographic separation system, for the unequivocal identification and structural elucidation of an unknown compound in the field of designer drugs. In a patient sample set (blood, tissues, vitreous humor, etc.), analyzed with a dedicated liquid chromatographic-fluorescence detection method for the determination of methylenedioxy amphetamine, methylenedioxy methamphetamine, and methylenedioxy ethylamphetamine (MDEA), a "strange" inexplicable peak appeared at a retention time not corresponding to any of our reference materials. Based on the identical excitation and emission wavelengths in detection, and a retention behavior comparable to MDEA, it was assumed that this unknown compound was an isomer of the recreational drug MDEA. With a simple and straightforward methodological crossover between LC fluorescence detection and LC-MS/MS, additional information for structural elucidation was easily obtained. Chromatographic separation was achieved on a Hypersil BDS C18 column (fluorescence detection part) and on a Hypersil BDS phenyl column (mass spectrometric detection part). MS showed that the unknown compound's molecular mass was identical to that of MDEA, and, in addition, its fragmentation pattern too proved quite similar to that of MDEA. A thorough literature overview and study of the fragmentation pattern by means of the MS/MS spectrum led to an evidence-based hypothesis of 3,4-methylenedioxy N,N-dimethylamphetamine (MDDM) being the unknown compound. To confirm this hypothesis, MDDM was synthesized and its presence in our biological sample was finally demonstrated by co-injection with alternatively synthesized MDDM and MDEA. This application shows the synergism between LC and MS in the elucidation of unknown compounds, nevertheless emphasizing the essence of chromatographic separation when dealing with isomers.

Measurement of plutonium isotope ratios in nuclear fuel samples by HPLC-MC-ICP-MS

Radioactive isotopes are traditionally quantified by means of radioactivity counting techniques (α, β, γ). However, these methods often require extensive matrix separation and sample purification before the identification of specific isotopes and their relative abundance is possible as it is necessary in the frame of post-irradiation examinations on nuclear fuel samples. The technique of multicollector inductively coupled plasma mass spectrometry (MC-ICP-MS) is attracting much attention because it permits the precise measurement of the isotope compositions for a wide range of elements combined with excellent limits of detection due to high ionization efficiencies. The present paper describes one of the first applications of an online high-performance liquid chromatographic separation system coupled to a MC-ICP-MS in order to overcome isobaric interferences for the determination of the plutonium isotope composition and concentrations in irradiated nuclear fuels. The described chromatographic separation is sufficient to prevent any isobaric interference between 238Pu present at trace concentrations and 238U present as the main component of the fuel samples. The external reproducibility of the uncorrected plutonium isotope ratios was determined to be between 0.04 and 0.2% (2 s) resulting in a precision in the ‰ range for the isotopic vectors of the irradiated fuel samples.

Separation and identification of compounds in Rhizoma chuanxiong by comprehensive two-dimensional liquid chromatography coupled to mass spectrometry

A comprehensive two-dimensional liquid chromatographic separation system based on the combination of a CN column and an ODS column is developed for the separation of components in a traditional Chinese medicine (TCM) Rhizoma chuanxiong. Two columns are coupled by a two-position, eight-port valve equipped with two storage loops and controlled by a computer. The effluent is detected by both the diode array detector and atmospheric pressure chemical ionization (APCI) mass spectrometer. More than 52 components in the methanol extract of R. chuanxiong were resolved and 11 of them were preliminary identified according to their UV and mass spectra.

Startup of transactinide chemistry in JAERI

The transactinide nuclides261Rf and262Db have been successfully produced in the248Cm(18O,5n) reaction at 99 MeV and in the248Cm(19F,5n) reaction at 100, 103, and 106 MeV, respectively, at the JAERI tandem accelerator. The on-line ion exchange experiments with an automated fast and repetitive liquid chromatography separation system were performed in the HNO3/HF system using Rf homologues89mZr and167,165Hf produced in the89Y(p,n) and152Gd(18O,xn) reactions, respectively. The radiotracers88Zr,175Hf, and234Th were also prepared and the distribution coefficients on ion exchange resins were measured systematically in 1-11 M HCl and 1-14 M HNO3with the batch method. It was found that anion exchange experiments of Rf in 8 M HNO3and 9 M HCl provided information useful to extract the ionic radius of Rf and to verify the influence of relativistic effects.

Rapid alcohol determination in plasma and urine by column liquid chromatography with biosensor detection

An enzyme based amperometric biosensor used as a selective and sensitive detection unit in column liquid chromatography for the determination of ethanol and methanol in biological fluids such as plasma and urine is described. The reagentless enzyme electrode is based on the co-immobilisation of alcohol oxidase and horseradish peroxidase in carbon paste. The selectivity of the biosensor was found to vary when four various alcohol oxidase enzyme preparations from Candida boidinii, Pichia pastoris, and Hansenula polymorpha were used in the biosensors described. High sensitivity could be obtained for a number of alcohols, organic acids, and aldehydes. Optimisation regarding the sensitivity and selectivity of the four alcohol oxidase co-immobilised biosensors are outlined. A fast and reliable liquid chromatographic separation system with a PLRP-S polymer based separation column used with a phosphate buffer as the mobile phase was optimised using the best biosensor which was based on alcohol oxidase from P. pastoris and which showed the highest turnover rate for alcohols, as the detector for the determination of ethanol and methanol in human urine and plasma samples. The selectivity and stability of the biosensor were retained by working at an applied potential of −50 mV versus Ag/AgCl, the optimal operational potential, and by the casting of a protective membrane on the electrode surface. High selectivity of the enzyme electrode was also found towards other easily oxidisable interfering species normally present in biological fluids. It was found that stable and reliable determinations of ethanol and methanol in plasma and urine could be performed with only a simple dilution and centrifugation step prior to injection into the liquid chromatographic system. An analysis time of 4 min was required for the assay, with a sample throughput of 13 samples h −1.

An automated liquid chromatographic plasma analysis of amino acids used in combination with positron emission tomography (PET) for determination of in vivo plasma kinetics

Quantification of physiological processes measured with positron emission tomography (PET) requires an “input” function which can be the concentration of administered radio-tracer in plasma. Radioactive nuclides used in PET have short half lives (2–20 min) and a limited time is available for the PET investigation including analysis of the composition of the radioactive signal in plasma. Therefore, an automated method for the analysis and separation of β-[ 11C]- l-5-hydroxy-tryptophan ([ 11C]- l-5-HTP), β-[ 11C]- l-3,4-dihydroxy-phenylalanine ([ 11C]- l-DOPA) and l-[methyl- 11C]-methionine and their respective metabolites in plasma was developed. A size exclusion column was used for isolation of the low molecular weight fraction. In the case of [ 11C]- l-5-HTP and [ 11C]- l-DOPA, the low molecular weight fraction was injected onto a liquid chromatographic system for separation of radioactive tracer from in vivo formed radio-labelled metabolites. The elution volume from the size exclusion column was 7.0, 5.0 and 3.5 ml for [ 11C]- l-5-HTP, [ 11C]- l-DOPA and l-[methyl- 11C]-methionine, respectively. An interaction with the column matrix and the solutes was observed for both [ 11C]- l-5-HTP and [ 11C]- l-DOPA. The yield in the isolation step was >98%. Separation of [ 11C]- l-5-HTP and [ 11C]- l-DOPA from their respective metabolites was performed with high-performance liquid chromatography with automated collection of fractions of the eluate corresponding to those of administered tracer and metabolites. The fractions were measured for radioactivity in a well counter. Inter- and intraday variation in retention were less than 3% RSD. Fortyfive minutes after injection of [ 11C]- l-5-HTP or [ 11C]- l-DOPA the radioactivity in the [ 11C]- l-5-HTP or [ 11C]- l-DOPA fraction was 58.5 ± 6.4 (RSD, n = 4) and 31.0 ± 2.7 (RSD, n = 3), respectively. High and low molecular weight fractions from plasma after injection of l-[methyl- 11C]-methionine were collected and measured for radioactivity. An incorporation of [ 11C]- l-methionine in plasma proteins, i.e. the high molecular weight fraction, was observed but with large inter-individual differences.

Human peripheral lymphocytes as indicators of microsomal epoxide hydrolase activity in liver and lung.

In this study, we have applied an improved assay for the determination of microsomal epoxide hydrolase activity to assess enzymatic levels in human lung, liver, and blood lymphocytes. The assay is fluorescence-based and monitors the epoxide hydrolase-mediated conversion of (+/-)-benzo[a]pyrene-4,5-epoxide to (+/-)-trans-benzo[a]pyrene-4,5-dihydrodiol, using a high pressure liquid chromatography separation system. Approximately a 40-fold range in microsomal epoxide hydrolase activities was detected in blood lymphocytes collected from 70 individual donors. In 38 individuals who were sampled twice after a 3-month interval, the repeatability of an individual's lymphocyte epoxide hydrolase activity was highly correlated (r = 0.80, p < 0.02). In addition, within the same individual there appeared to be a strong correlation between lymphocyte and liver epoxide hydrolase activity (r = 0.92, p = 0.02), and some correlation between liver and lung activity (r = 0.58, p = 0.05). Activities were assessed in lymphocytes from a styrene-exposed worker population but no significant associations between blood concentrations of styrene and epoxide hydrolase activity levels were observed. Neither were any correlations detected in these workers between epoxide hydrolase activities and age, years on the job, alcohol consumption, sex, or smoking status. The results of our study suggest that blood lymphocytes are a useful sentinel cell for epoxide hydrolase activity determinations in individuals, as these measures are relatively stable over time and appear to reflect activity levels in other target organs.

Fully automated on-line liquid chromatographic separation system for polar pollutants in various types of water

A fully automated column liquid chromatographic separation system using on-line trace enrichment, gradient elution and diode-array detection for the trace-level determination of polar pollutants is described. Automation of the system was achieved by means of an automated cartridge-exchange system (PROSPEKT). Relevant parameters such as pH, volume and ionic strength of the sample, flow-rate during the enrichment step and wavelengths and band widths during detection were optimized for eighteen pollutants in various types of water at concentration levels below 5 μg l −1. The determination limit for all test compounds in liquid chromatographic grade water was 0.1 μg l −1, and identification, via diode-array spectra, could be performed at the same level. The mean relative standard deviations of the peak areas and the retention times for all the test compounds were 10% and 0.3%, respectively, at the 5 μg l −1 level for river Rhine water.

Characterization of biotechnological processes and products using high‐performance liquid chromatography (H P L C). VII. Determination of organic compounds in sulphite liquor

AbstractCarbohydrates, acetic acid, furfural, 5‐hydroxymethylfurfural (HMF) were analyzed in sulphite liquor using HPLC with refractive index detection.Pentoses (xylose, arabinose) and hexoses (glucose, galactose, mannose) were separated on an Aminex HPX‐87P column as wel as by means of a self‐made glass cartridge packed with a piperazine impregnated silica gel. Before injecting into the HPLC apparatus the sulphite liquors were purified using a presep column packed with a cation‐exchange resin. The sugar quantification between the two different separation systems was satisfactory.Using an Aminex HPX‐87P column and an UV monitor set at 283 nm connected in parallel with the RI detector also furfural and HMF could be analyzed in sulphite liquor.Furthermore, an Aminex HPX‐87H column and an UV detector set at 210 nm were most suitable to quantify ethanol.These modified and optimized liquid chromatographic separation systems are the basis for further investigations of the utilization of hexoses as well as pentoses in sulphite liquors by means of suitable microorganisms.

Fibre optic sensors for the characterization of particle size and flow velocity

Photon correlation spectroscopy (PCS) has been applied to the routine monitoring of proteins eluting off liquid chromatographic separation systems. The technique has been configured as an on-line, in situ detector capable of remote analyses using fibre optic technology. Analysis of flowing protein samples has been demonstrated and its use in process biochemistry as a quality assurance monitor is discussed. The bulk optics, gas lasers and sophisticated detection systems found in conventional systems have been replaced with miniaturized solid-state components for incorporation into a fibre optic multiplexable (LANS) network for multipoint analyses at remote points in the non-laboratory environment. A fibre optic PCS system has been shown to analyse protein samples with close to theoretical maximum efficiency, permitting significant reductions in incident light powers to be achieved. The determination of flow velocity in more than one dimension is conventionally carried out using complex and expensive bulk optical systems. A fibre optic version, employing a single fibre pair, has been designed that will permit characterization of flow velocity profiles, independent of direction, for use as a multiplexable network capable of measurements in situations previously inaccessible to such analyses. The passive sensor head requires no alignment or adjustment and contains specially coated lenses on an individual fibre for generation of the required optical scattering volume.

Microcomputer-assisted liquid chromatographic separation system (mcasyst) for method development and data handling

A computer-assisted liquid chromatographic separation system (MCASYST) has been developed for method development and related data handling, focusing especially on automated identification of separated components. The system has six main functions: retention prediction system, liquid chromatographic database system, automated identification system, automated optimization system for separation conditions, data loading program from a UV multi-channel detector and UV spectral database system. The performance and potential of the MCASYST system are discussed with respect to the analysis of phenylthiohydantoin-amino acids.