Anion-exchange Liquid Chromatography Research Articles

Using QuEChERS sample preparation technique followed by ICP-Ms method to determination Cr(III) and Cr(VI) in seawater

Currently, Cr(III) and Cr(VI) are analyzed directly in trace level using anion-exchange liquid chromatography inductively coupled plasma mass spectrometry (LC – ICP – MS), which is well-known as an expensive and complicated method. Therefore, determination of Cr(VI) and Cr(III) by spectrometer or combine with ICP (or AAS) is seemly prefer to choose in almost laboratories although this method presents a higher limit of detection (LOD) and is influenced by salt ions in seawater. In this study, after forming reddish-violet complex by adding diphenylcarbazide (DPC) to seawater samples, a QuEChERS extraction technique was applied to separate Cr from high salt level matrix and transfer the Cr(VI)-DPC structure to the acetonitrile phase. In case of determination of total Cr, the Cr(III) in samples had to convert to Cr(VI) by oxidate with KMnO4 before adding DPC. Then, a digestion process with HNO3 – H2O2 mixture was implemented before analyzing Cr(VI) by ICP-MS. The Cr(III) is calculated indirectly based on total Cr and Cr(VI) results. This method had a linear concentration range of Cr(VI) from 0.02 to 50 µg/L with a detection limit of 0.015µg/L, the relative standard deviations under 3,03%, and the recovery was 92-108 % for seawater. According to the outcome of parallel experiment with common method, which used spectrometer and did not apply QuEChERS extraction step, the ICP-MS method presented a better result with smaller LOD, RSD, and recovery range.

Direct, automated and sensitive determination of glyphosate and related anionic pesticides in environmental water samples using solid-phase extraction on-line combined with liquid chromatography tandem mass spectrometry

An automated procedure for the simultaneous determination of six anionic pesticides, including glyphosate (GLY) and its transformation product aminomethylphosphonic acid (AMPA), was developed and applied to the analysis of environmental water samples. The proposed method combines on-line concentration of water samples (0.160 mL), with compounds separation in an anion-exchange liquid chromatography (LC) column, followed by their selective determination by tandem mass spectrometry (MS/MS). The global procedure was completed in 25 min, providing limits of quantification (LOQs) between 5 ng L−1 and 20 ng L−1, with reduced effect of the surface water matrix in the efficiency of process (SPE and ionization yields). The method was applied to the analysis of grab samples obtained from three watersheds, in two rural and one residential area, in Galicia (Northwest Spain). Out of six investigated compounds, Fosetyl, AMPA and GLY were noticed in the set of processed samples. Their detection frequencies increased from 12% (Fosetyl) to 88% (AMPA). Median concentrations followed the same trend varying from 9 ng L−1 (Fosetyl) to 44 ng L−1 (AMPA). The higher levels and the large seasonal variations in the residues of the latter species were noticed in small rivers affected by discharges of municipal sewage treatment plants (STPs).

Chemical Fruit Profiles of Different Raspberry Cultivars Grown in Specific Norwegian Agroclimatic Conditions

Raspberries are considered valuable fruits due to their high levels of nutrients and phytochemicals, which have many beneficial effects on humans. As many external factors affect the composition of these fruits (the type of cultivation, soil characteristics, ripeness, storage time and post-harvest technologies, cultivar/genotype, and climatic conditions), the goal of this study was to analyze different raspberry cultivars grown in Norway. Considering that Norway is a country with specific climatic conditions, as well as has a limited period of fruit vegetation, another important goal of this study was also to compare raspberries from different Norwegian areas, as well as different grown cultivars. Modern analytical techniques, such as high-performance anion-exchange liquid chromatography with pulsed amperometric detection (HPEAC-PAD), ultra-high-performance liquid chromatography with diode array detector coupled to triple quadrupole mass spectrometry (UHPLC-DAD MS/MS), and inductively coupled plasma–optical emission spectrometry (ICP-OES), provided a detailed examination of the raspberry extract samples. Based on their high levels of minerals (especially N, P, and K), organic acids (predominantly citric and malic acids), sugars (glucose, fructose, sucrose, and galactose), and polyphenols (ellagic acid, syringic acid, quercetin, and rutin), Norwegian raspberries could be considered fruits with increased health-beneficial compounds. The chemical composition of the studied cultivars depended on the locality of growth.

Speciation of Arsenic(III) and Arsenic(V) in Plant-Based Drinks.

Recently, food products based only on plants have become increasingly popular and are often found on store shelves. It is a specific market response to the growing demand for, and interest in, plant foods. Cow’s milk has also gained its counterpart in the form of plant-based beverages, based on cereals, nuts or legumes. The emergence of an increasingly wide range of plant-based food products has also led to increased research on safe plant food consumption. This study was conducted to quantify total arsenic content and its species (arsenic(III) and (V)) in samples of plant-based beverages purchased at Polish markets. Speciation analysis of arsenic was performed by high-performance liquid chromatography combined with inductively coupled plasma mass spectrometry. The presented study was conducted on six selected plant-based beverages, including almond, millet, soybean, rice, coconut and oat. An analysis using size exclusion chromatography was performed. In order to initially visualize the content of the observed elements and the particle size of the compounds in which they occur, at first the samples were subjected to the size-exclusion chromatography. Speciation analysis of arsenic was carried out using anion-exchange liquid chromatography, combined with inductively coupled plasma mass spectrometry. The presented method was validated with certified reference material (CRM rice flour).

Combination of strong anion exchange liquid chromatography with microchip capillary electrophoresis sodium dodecyl sulfate for rapid two-dimensional separations of complex protein mixtures

Two-dimensional separations provide a simple way to increase the resolution and peak capacity of complex protein separations. The feasibility of a recently developed instrumental approach for two-dimensional separations of proteins was evaluated. The approach is based on the general principle of two-dimensional gel electrophoresis. In the first dimension, semi-preparative strong anion exchange high-performance liquid chromatography is utilized and fractions are collected by means of a fraction collector. They are subsequently analyzed in the second dimension with microchip capillary electrophoresis sodium dodecyl sulfate. Microchip capillary electrophoresis provides the necessary speed (approximately 1 min/fraction) for short analysis. In this study, three different samples were investigated. Different constructs of soluble guanylyl cyclase were expressed in Sf9-cells using the baculovirus expression system. Cell lysates were analyzed and the resulting separations were compared. In our experimental setup, the soluble guanylyl cyclase was identified among hundreds of other proteins in these cell lysates, indicating its potential for screening, process control, or analysis. The results were validated by immunoblotting. Samples from Chinese hamster ovary cell culture before and after a purification step were investigated and approximately 9% less impurities could be observed. The separation patterns obtained for human plasma are closely similar to patterns obtained with two-dimensional gel electrophoresis and a total of 218 peaks could be observed. Overall, the approach was well applicable to all samples and, based on these results, further directions for improvements were identified.Graphical abstract.

Innovatively Established Analysis Method for Lipoprotein Profiles Based on High-Performance Anion-Exchange Liquid Chromatography.

Separation analysis of lipoprotein classes have various methods, including ultracentrifugation, electrophoresis, and gel permeation chromatography (GPC). All major lipoprotein classes can be separated via ultracentrifugation, but performing the analysis takes a long time. Low-density lipoprotein (LDL), intermediate-density lipoprotein (IDL), and very low-density lipoprotein (VLDL) in patient samples cannot be sufficiently separated via electrophoresis or GPC. Thus, we established a new method [anion-exchange high-performance liquid chromatography (AEX-HPLC)] by using HPLC with an AEX column containing nonporous gel and an eluent containing chaotropic ions. AEX-HPLC can separate five lipoprotein fractions of high-density lipoprotein (HDL), LDL, IDL, VLDL, and others in human serum, which can be used in substitution for ultracentrifugation method. The method was also approved for clinical use in the public health-care insurance in Japan in 2014. Furthermore, we developed an additional method to measure cholesterol levels of the four leading lipoprotein fractions and two subsequent fractions (i.e., chylomicron and lipoprotein(a)). We evaluated the clinical usefulness of AEX-HPLC in patients with coronary heart disease (CHD), diabetes, and kidney disease and in healthy volunteers. Results indicate that the cholesterol levels in IDL and VLDL measured by AEX-HPLC may be useful risk markers of CHD or diabetes. Furthermore, we developed another new method for the determination of alpha-tocopherol (AT) in lipoprotein classes, and this method is composed of AEX-HPLC for the separation of lipoprotein classes and reverse-phase chromatography to separate AT in each lipoprotein class. The AT levels in LDL were significantly correlated with the lag time to copper ion-induced LDL oxidation, which is an index of oxidation resistance. The application of AEX-HPLC to measure various substances in lipoproteins will be clinically expected in the future.

Analytical methodology for the simultaneous determination of NMG-Sb(v), iSb(v), and iSb(iii) species by anion exchange liquid chromatography in Glucantime® and its biological application in Wistar rat urine

An analytical method for the determination of Sb species in Glucantime® and rat urine samples.

PBN11-8, a Cytotoxic Polypeptide Purified from Marine Bacillus, Suppresses Invasion and Migration of Human Hepatocellular Carcinoma Cells by Targeting Focal Adhesion Kinase Pathways.

The development of antitumor drugs has attracted cancer researchers and the identification of novel antitumor lead compounds is certainly of great interest. The fermentation broth of Bacillus sp. N11-8, which was isolated from the Antarctic waters, showed cytotoxicity towards different cells. A cytotoxic polypeptide, PBN11-8, was purified from the fermentation broth of Bacillus sp. N11-8 using ultrafiltration, ammonium sulfate precipitation, anion exchange liquid chromatography and high performance liquid chromatography (HPLC). Cloning and sequence analysis showed that PBN11-8 polypeptide (MW: ~19 kDa by the electrospray-ionization (ESI)) displayed high similarity with peptidase M84 from Bacillus pumilus. PBN11-8 possessed moderate cytotoxicity towards several cancer cell lines with IC50 values of 1.56, 1.80, 1.57, and 1.73 µg/mL against human hepatocellular carcinoma cell line BEL-7402, human renal clear cell adenocarcinoma cell line 786-0, human hepatocellular carcinoma cell line HepG2, and human pancreatic cancer cell line Panc-28, respectively. Moreover, the polypeptide displayed weak cytotoxicity towards normal cell line renal tubular epithelial cell line HK2 and human normal liver cell line L02 cells. Wound healing migration and Transwell experiments demonstrate that PBN11-8 could inhibit the migration and invasion of BEL-7402. Further investigation revealed that PBN11-8 suppresses focal adhesion kinase (FAK)-mediated adhesion, migration, and invasion by disturbing FAK/extracellular regulated protein kinases (ERK) signaling and matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9 (MMP-9) in BEL-7402 cells. Thus, PBN11-8 represents a potential novel anti-cancer lead compound.

Dual Electrolytic Eluent Generation for Oligosaccharides Analysis Using High-Performance Anion-Exchange Chromatography.

The research on oligosaccharides is growing and gaining in importance at a rapid pace. The efforts to understand their bioactivity and to develop new products based on oligosaccharides in biotherapeutics and food industry require effective and reliable tools for analysis of oligosaccharides. Here we present a dual electrolytic eluent generation platform for the analysis of oligosaccharides by high-performance anion-exchange liquid chromatography (HPAE) in both analytical and capillary column formats. The system consists of one eluent generator producing methanesulfonic acid (MSA) connected in series with a second eluent generator producing potassium hydroxide (KOH). Through manipulating the concentration output of both eluent generators, chromatographic performance comparable to that obtained using the conventional sodium acetate/sodium hydroxide (NaOAc/NaOH) eluents is achieved using the electrolytically generated potassium methanesulfonate/potassium hydroxide (KMSA/KOH) eluent. This platform utilizes deionized water as the only carrier stream through a single isocratic pump, overcomes the various drawbacks associated with manually prepared NaOAc/NaOH eluents, and offers an easy to use, simplified operation solution for oligosaccharides profiling with increased precision and accuracy.

Speciation analysis of arsenic in seafood and seaweed: Part II-single laboratory validation of method.

Single laboratory validation of a method for arsenic speciation analysis in seafood and seaweed is presented. The method is based on stepwise extraction of water-soluble and non-polar arsenic with hot water and a mixture of dichloromethane and methanol, respectively. While the water-soluble arsenicals were speciated by anion and cation exchange liquid chromatography inductively coupled plasma mass spectrometry (LC-ICP-MS), the non-polar arsenicals were collectively determined by ICP-MS after digestion in acid. The performance characteristics and broad application of the method were evaluated by analyzing eight commercial samples (cod, haddock, mackerel, crab, shrimp, geoduck clam, oyster, and kombu) and four reference materials (fish protein (DORM-4), lobster hepatopancreas (TORT-3), mussel tissue (SRM 2976), and hijiki seaweed (CRM 7405-a)) representing finfish, crustaceans, molluscs, and seaweed. Matrices spiked at three levels in duplicates were also analyzed. The stepwise extraction provided 76-106% extraction of the total arsenic from the test materials. The method demonstrated satisfactory repeatability for analysis of replicate extracts prepared over several days. The accuracy of the method was evaluated by analyzing reference materials certified for both total arsenic and a few arsenicals; the experimental results were 90-105% of the certified values. Comparison between the total water-soluble arsenic and the sum of the concentrations of the chromatographed species gave 80-92% mass balance. While spike recoveries of most arsenicals were in the acceptance range set by CODEX, a few species spiked into cod, haddock, and shrimp were poorly recovered due to transformation to other forms. After thorough investigations, strategies were devised to improve the recoveries of these species by averting their transformations. Limits of quantification (LOQ) for the extraction and quantification of 16 arsenicals using the current method were in the range 6-16ng g-1 arsenic.

Analysis of Main Arsenic Species in Canned Fish Marketed in the Czech Republic and Austria

Total arsenic and water-soluble arsenic species were determined in canned fish bought in Graz (n = 6) and Prague (n = 8) with inductively-coupled plasma mass spectrometry and anion-exchange liquid chromatography. A difference in total arsenic content between samples from Prague (mean 3.4; standard deviation (s.d.) 1.2 mg kg–1) and Graz (mean 2.5; s.d. 0.7 mg kg–1) was not significant, however Prague samples differed in abundance of arsenobetaine (mean 83% of extractable arsenic; s.d. 16%) vs. Graz samples (mean 66%; s.d. 10%). No differences in total arsenic were caused by fish species (mackerel, sardine, herring, tuna) and origin (Northeast and Eastern-central Atlantic).

Effect of dietary modification by calorie restriction on cholesterol levels in lipoprotein(a) and other lipoprotein classes.

Background Dietary habits are associated with obesity which is a risk factor for coronary heart disease. The objective is to estimate the change of lipoprotein(a) and other lipoprotein classes by calorie restriction with obesity index and Framingham risk score. Methods Sixty females (56 ± 9 years) were recruited. Their caloric intakes were reduced during the six-month period, and the calorie from fat was not more than 30%. Lipoprotein profiles were estimated at baseline and after the six-month period of calorie restriction. Cholesterol levels in six lipoprotein classes (HDL, LDL, IDL, VLDL, chylomicron and lipoprotein(a)) were analysed by anion-exchange liquid chromatography. The other tests were analysed by general methods. Additionally, Framingham risk score for predicting 10-year coronary heart disease risk was calculated. Results Body mass index, waist circumference, insulin resistance, Framingham risk score, total cholesterol, LDL-cholesterol and IDL-cholesterol were significantly decreased by the calorie restriction, and the protein and cholesterol levels of lipoprotein(a) were significantly increased. The change of body mass index was significantly correlated with those of TC, VLDL-cholesterol and chylomicron-cholesterol, and that of waist circumference was significantly correlated with that of chylomicron-cholesterol. The change of Framingham risk score was significantly correlated with the change of IDL-C. Conclusion Obesity indexes and Framingham risk score were reduced by the dietary modification. Lipoprotein profile was improved with the reduction of obesity indexes, but lipoprotein(a) was increased. The changes of obesity indexes and Framingham risk score were related with those of triglyceride-rich lipoproteins, e.g. IDL, VLDL and CM.

Purification and characterization of trypsin from Luphiosilurus alexandri pyloric cecum

Trypsin from L. alexandri was purified using only two purification processes: ammonium sulfate precipitation and anion exchange liquid chromatography in DEAE-Sepharose. Trypsin mass was estimated as 24kDa through SDS-PAGE, which showed only one band in silver staining. The purified enzyme showed an optimum temperature and pH of 50°C and 9.0, respectively. Stability was well maintained, with high levels of activity at a pH of up to 11.0, including high stability at a temperature of up to 50°C after 60min of incubation. The inhibition test demonstrated strong inhibition by PMSF, a serine protease inhibitor, and Kinetic constants km and kcat for BAPNA were 0.517mM and 5.0S−1, respectively. The purified enzyme was also as active as casein, as analyzed by zymography. Therefore, we consider trypsin a promising enzyme for industrial processes, owing to its stability in a wide range of pH and temperature and activity even under immobilization.

Online eluent-switching technique coupled anion-exchange liquid chromatography–ion trap tandem mass spectrometry for analysis of non-steroidal anti-inflammatory drugs in pig serum

A novel method for online extraction, pH-gradient separation, and analysis of nine non-steroidal anti-inflammatory drugs (NSAIDs) was developed by coupling online eluent-switching technique to single anion-exchange chromatographic column/ion trap mass spectrometer (MS) and used for monitoring NSAIDs residues in pig serum. A neutral eluent and a pH-gradient eluent were used for extraction and separation of NSAIDs, respectively. Each of nine NSAIDs has an MS precursor ion of either [M−H]− or [M−Na]−. The extracted ion chromatogram for a specific product ion of each NSAID was used for its quantitative analysis. The dynamic linear ranges of calibration curves were all 0–200ngmL−1 (R2>0.9950). The analysis accuracies estimated by spiking standard concentrations at 20, 100, and 200ngmL−1 were 80.5–99.9%. The corresponding intra-day and inter-day precisions (RSD%) were 2.5–14.5% and 2.9–15.2%, respectively. The limit of detection/limit of quantitation of NSAIDs were 1.3/4.3, 0.5/1.6, 0.2/0.5, 2.5/8.2, 1.5/4.9, 0.6/2.1, 0.6/2.0, 0.5/1.7, and 0.6/2.1ngmL−1 for carprofen, diclofenac, flunixin, ibuprofen, ketoprofen, meclofenamic acid sodium, mefenamic acid, niflumic acid, and tolfenamic acid, respectively. After 1h injection of a dose containing 2mgkg−1 weight pig of flunixin and tolfenamic acid to the pigs, a residue amount of 3480±36ngmL−1 and 431±13ngmL−1, respectively, was reached for the incurred pig serum specimens and both residues were reduced to about 20ngmL−1 at the time of 24h.

Determination of Methamphetamine and Amphetamine in Hair by Mixed-Mode Anion Exchange Solid Phase Extraction and Liquid Chromatography–Tandem Mass Spectrometry

Methamphetamine is one of the most widely used sympathomimetic amines in Asia. To identify cases of methamphetamine abuse and to estimate its abuse, hair segmental analysis is used extensively. However, existing hair analysis has several limitations due to matrix interferences. In an attempt to resolve these issues, a simple, rapid, and sensitive liquid chromatography–tandem mass spectrometry method is reported for the determination of methamphetamine and amphetamine in human hair with anion exchange solid phase extraction. Hair dissolved in sodium hydroxide was loaded on a mixed-mode anion exchange cartridge and amphetamine and methamphetamine were eluted with methanol in less than ten minutes followed by liquid chromatography–tandem mass spectrometry. The recovery of this method was between 93.9 and 102.0 percent. The limits of detection and quantitation were 0.01 and 0.05 nanograms per milligram for both analytes. The accuracy and precision were satisfactory for quantitative analysis. The suitability of the method for practical analysis was investigated using hair from methamphetamine users.

Development and validation of a stability indicating method for S-carboxymethyl-l-cysteine and related degradation products in oral syrup formulation

A stability-indicating method for the determination of S-carboxymethyl-l-cysteine and related degradation impurities in Exputex® 250mg/5mL syrup was developed in anion-exchange liquid chromatography mode. A forced degradation study supported the method development to ensure stability indicating conditions. Aqueous solutions of the active pharmaceutical ingredient and syrup samples at different pH-values were stress-tested in different thermal, light exposure and headspace conditions. One degradation product was detected in thermal stress studies at 60°C and 80°C in the pH range 5.0–7.0 and was identified by mass spectrometry as 5-oxo-thiomorpholine-3-carboxylic acid (lactam of carbocysteine). A second degradation product was only generated in moderately strong oxidizing conditions (0.5% H2O2 aqueous solution) and was identified as S-carboxymethyl-l-cysteine-(R/S)-sulphoxide (carbocysteine sulphoxide).The method was developed on a Zorbax SAX column, in isocratic mode. The mobile phase consisted of 200mM phosphate solution at pH 4.0 and acetonitrile (50:50 v/v) and UV detection was performed at a wavelength of 205nm. The method was linear for carbocysteine (R>0.9982) over a concentration range of 2.5–50μg/mL and 0.4–0.6mg/mL. Linearity for the impurities was shown from the LOQ to 50μg/mL. Specificity was verified and accuracy demonstrated for the active ingredient and its degradation products in syrup samples at 3 levels around their respective specification limits. Repeatability, intermediate precision and inter-laboratory reproducibility were assessed on three commercial batches, analyzed in triplicate by two operators at both the transferring and the receiving site and demonstrated a successful method transfer to the manufacturing quality control laboratory.

Simultaneous speciation and determination of arsenic, chromium and cadmium in water samples by high performance liquid chromatography with inductively coupled plasma mass spectrometry

The simultaneous separation and determination of As(iii), As(v), monomethylarsenic acid (MMA), dimethylarsenic acid (DMA), Cr(iii), Cr(vi) and Cd(ii) in water samples have been carried out by anion exchange liquid chromatography with inductively coupled plasma mass spectrometry.

Development and validation of an ion-exchange chromatography method for heparin and its impurities in heparin products.

An anion-exchange liquid chromatography method for the determination of heparin and its impurities (dermatan sulfate and oversulfated chondroitin sulfate) was developed using chemometric-assisted optimization, including multivariate experimental design and response surface methodology. The separation of heparin, dermatan sulfate, and oversulfated chondroitin sulfate (Rs above 2.0) was achieved on a Dionex RF IC IonPac AS22 column with a gradient elution of 10-70% of 2.5M sodium chloride and 20mM Tris phosphate buffer (pH 2.1) at a flow rate of 0.6mL/min and UV detection at 215nm. Method validation shows good linearity (r > 0.99), acceptable precision (%relative standard deviations <11.4%) and trueness (%recovery of 92.3-103.9%) for all analytes. The limits of detection for dermatan sulfate and oversulfated chondroitin sulfate are equivalent to 0.11% w/w (10.5μg/mL) and 0.07% w/w (7.2μg/mL), while the limits of quantification are 0.32% w/w (31.5μg/mL) and 0.22% w/w (22.0μg/mL) relative to heparin, respectively. The method is specific for heparin, dermatan sulfate, and oversulfated chondroitin sulfate without interference from mobile phase and sample matrices and could be used for accurate quantitation the drug and its impurities in a single run. Applications of the method reveal contents of heparin between 90.3 and 97.8%. Dermatan sulfate and oversulfated chondroitin sulfate were not detected in any of the real-life samples.

Separation and Purification of Sinigrin and Gluconapin from Defatted Indian Mustard Seed Meals by Macroporous Anion Exchange Resin and Medium Pressure Liquid Chromatography

Defatted seed meals, the byproducts of Indian mustard seed oil industry, which are just used as animal feedstock and nitrogen fertilizers, possess various nutrients and phytochemical compounds, such as sinigrin and gluconapin. The present study demonstrated a novel, low-cost, and efficient method for the co-production of sinigrin and gluconapin from defatted Indian mustard seed meals by D261 strong basic anion-exchange macroporous resin and medium pressure liquid chromatography (MPLC). The adsorption/desorption capacities of eight adsorbents characterized by BET, IR, and EDS were screened. 26.4 g defatted Indian mustard seed meals, which contained about 1210.3 mg sinigrin and 696.6 mg gluconapin, could produce 2059.6 mg of glucosinolate-rich extract of 53.67% sinigrin and 31.30% gluconapin after separation by D261 resin. Then, 949.40 mg of 97% sinigrin and 568.40 mg of 95% gluconapin could be obtained after the extracts were purified by MPLC, with the recovery of 76.03% of sinigrin and 77.38% of gluconapin. The products were assessed by analytical HPLC and characterized by UV, MS, 1H NMR, and 13C NMR. In conclusion, this method is practical and environmentally friendly, and it is a low-cost process to make full use of Indian mustard seeds.

Phosphorus-specific determination of glyphosate, glufosinate, and their hydrolysis products in biological samples by liquid chromatography–inductively coupled plasma–mass spectrometry

Anion exchange liquid chromatography (LC) with inductively coupled plasma–mass spectrometry (ICP–MS) was used for simultaneous determination of glyphosate and glufosinate (phosphonic and amino acid group-containing herbicides; PAAHs) and their hydrolysis products, aminomethylphosphonic acid (AMPA) and 3-methylphosphinicoacetic acid (MPPA), in biological samples. The target compounds were separated using an anion exchange resin column and gradient elution with sodium carbonate and sodium hydroxide system. The chromatographic eluates were passed through a membrane suppressor system and monitored by phosphorus-specific detection at m/z 31. PAAHs and their hydrolysis products were baseline separated from each other within 40 min of elution time, and phosphoric acid did not interfere with detection. The detection limits in serum samples were 0.1–0.7 µg/ml; those in urine samples, 0.2–1.6 µg/ml for the four compounds. The spiked recoveries for the four compounds were over 91 % in serum and urine samples. The detection of the compounds was not subject to interference from sample matrix components. The present method would be useful for toxicological analysis of PAAHs and their products in actual forensic practice. To our knowledge, this is the first trial to establish an LC–ICP–MS method for analysis of PAAHs and their products in biological samples .