Concentration Of Sodium Perchlorate Research Articles

Anion exchange-HPLC method for evaluating the encapsulation efficiency of mRNA-loaded lipid nanoparticles using analytical quality by design

Lipid nanoparticles (LNPs) are emerging nucleic acid delivery systems in the development of mRNA therapeutics such as the severe acute respiratory syndrome coronavirus 2 vaccines. However, a suitable analytical method for evaluating the encapsulation efficiency (EE) of the LNPs is required to ensure drug efficacy, as current analytical methods exhibit throughput issues and require long analysis times. Hence, we developed and validated an anion-exchange HPLC method using Analytical Quality by Design. Three critical method parameters (CMPs) were identified using risk assessment and Design of Experiments: column temperature, flow rate, and sodium perchlorate concentration. The CMPs were optimized using Face-Centered Central Composite Design. The discriminating power of the optimized HPLC method and RiboGreen assay was comparable. The main advantage of this method is that LNPs can be directly injected into the HPLC system without bursting the LNPs loaded with encapsulated poly(A). The optimized HPLC method was validated as robust, high-throughput, and sufficiently sensitive according to the ICH Q2 guidelines. We believe our findings could promote efficient LNPs-based drug development.

Thyroid-gonadal hormonal interplay in zebrafish exposed to sodium perchlorate: Implications for reproductive health

Perchlorate, a widespread environmental contaminant originating from various industrial applications, agricultural practices, and natural sources, poses potential risks to ecosystems and human health. While previous studies have highlighted its influence on the thyroid endocrine system and its impact on gonadal maturation, reproduction, and sex hormone synthesis, the specific interplay between thyroid and steroid hormones, in this context, remains largely unexplored. Therefore, this study was undertaken to investigate the adverse effects and underlying mechanisms triggered by exposure to sodium perchlorate (SP) on reproductive endocrine activity in zebrafish. For 21 d, the fish were exposed to test SP concentrations (0, 3, 30, 300 mg/L), which were determined based on the exposure concentrations that induced various toxic effects in the fish, considering naturally occurring concentrations. Exposure to SP, except at 3 mg/L in males, significantly decreased the production of thyroid hormone (TH) in both female and male zebrafish. Moreover, gonadal steroid levels were markedly reduced in both sexes. The expression of hepatic vitellogenin (VTG) mRNA in female zebrafish was significantly decreased, whereas aromatase activity in male zebrafish was significantly elevated in the SP exposure groups. The reduced levels of THs and gonadal steroid hormones were strongly correlated. Abnormal responses to SP exposure led to reduced reproductive success in the 300 mg/L SP exposure group. These findings indicate that prolonged and continuous exposure to a specific concentration of SP may lead to long-term reproductive problems in zebrafish, primarily through hormonal imbalances and suppression of hepatic VTG mRNA expression.

Monitoring contamination of perchlorate migrating along the food chain to dairy products poses risks to human health

Assessments of human exposure to sodium perchlorate via dairy sources are limited. The current study applied untargeted metabolomics (LOD, 1.08–35.60 μg L-1; LOQ, 2.54–90.58 μg L-1; RSD < 6.2%) and proteomics methods by UHPLC-Q-Orbitrap HRMS to investigate the metabolic pathways and nutritional quality of goat milk contaminated with sodium perchlorate. Specifically, 11 metabolites including lactose (from 2.01 to 0.58 mg L-1), adenosine 5′-monophosphate (from 1.23 to 0.45 mg L-1), hypoxanthine (from 0.63 to 0.08 mg L-1), etc. and 3 crucial enzymes include α-lactalbumin, xanthine dehydrogenase and creatine kinase related to the quality traits of goat milk after sodium perchlorate treatment. Overall, except for spermidine, other related metabolites significantly decreased with the increase of sodium perchlorate concentration 0–160 μg L-1 and storage time (4–12 h). Collectively, we provide previously uncharacterized goat milk nutritional quality degradation mechanism induced by sodium perchlorate and a reference to ensure its safe use in human health.

Electrochemical mineralization of iron-tannate stain on HAp and bovine enamel-A non-peroxide approach

Prolonged treatments for the destaining of teeth using high concentrations of hydrogen peroxide may cause secondary unwanted effects such as tooth hypersensitivity and gingival irritation. Hence, it is aimed to develop a non-peroxide-based method to oxidize iron-tannate (Fe-TA) stained hydroxyapatite (HAp) and bovine enamel (BE) samples. Constant current electrolysis (CCE) experiments were carried out on Pt working electrode in aqueous NaCl, KCl and KI solutions at discrete concentrations under continuous experiment and a non-continuous experiment. CCE shows that in the presence of iron tannate (Fe-TA) stained HAP, approximately 30 ppm of iodine was generated using 0.1M KI and nearly 40 ppm was produced with 0.2 M KI. By using a non-continuous CCE process, the lowest amount of chlorine was generated from NaCl solution, which was well within the safety limits for oral applications. Depending on the experimental conditions used, between 13 ppm and 124 ppm of chlorine was generated. CCE of Fe-TA stained on HAp using KCl reveals that at the lowest current density of 10 mA/cm2, the amount of hypochlorite generated was 20 ppm on Pt electrode having a surface area of 6 cm2. Ion chromatographic (IC) analysis revealed that non-continuous CCE of Fe-TA-BE in NaCl generated a low concentration of sodium perchlorate (0.8 ppm), whereas the continuous process generated no perchlorate, but a considerable higher quantity of chlorate for Fe-TA-BE (37 ppm) and Fe-TA-HAp (140 ppm) samples.

Template‐free electrochemically polymerized polypyrrole nanowires and their application in flexible solid‐state supercapacitors

AbstractTreated carbon cloths combined with polypyrrole (PPy) nanowires (TCC@PPyW) were fabricated by template‐free electrochemical deposition of pyrrole in the presence of sodium citrate dihydrate and sodium perchlorate. The morphologies and electrochemical performances of the samples were studied using scanning electron microscopy, cyclic voltammetry, galvanostatic charge–discharge measurements and electrochemical impedance spectroscopy. The morphologies of PPy nanostructure were controlled by regulating interfacial pH, deposition time and concentration of sodium perchlorate. PPy nanorods were generated with the aid of water oxidation when the pH of pyrrole solution was 7.9, and PPy nanowires could be obtained by increasing the deposition time or the concentration of sodium perchlorate. The resulting PPy nanowires were about 100 nm in diameter and up to 600 nm in length. TCC@PPyW electrodes provided a large contact area for electrolyte, which facilitated the rapid delivery of electric charge. As a result, the TCC@PPyW electrodes exhibited a high areal specific capacitance of 841.2 mF cm−2 at 0.4 mA cm−2 and a high energy density of 420.6 μWh cm−2 at a power density of 0.2 mW cm−2. An all‐solid‐state flexible symmetric supercapacitor without any binder, incorporated with TCC@PPyW as electrodes and poly(vinyl alcohol)–LiCl gel as gel electrolyte, is also reported. The TCC@PPyW supercapacitor displayed excellent flexibility and rate capability. © 2021 Society of Chemical Industry

Na0.44MnO2/Polyimide Aqueous Na-ion Batteries for Large Energy Storage Applications

Aqueous salt batteries with high concentrations of salt or water in salt aqueous systems have received considerable attention with focus on improving working voltage range and energy density. Here, the effect of NaClO4 salt concentration on the electrochemical performance and stability of tunnel-type Na0.44MnO2 (NMO) cathodes and organic polyimide (PI) derivative anodes was studied. High capacity retention and 100% coulombic efficiency were shown for NMO/PI full cell in saturated NaClO4 electrolyte. A high, stable capacity of 115 mAh/g was achieved for the PI anode material, and the full cell showed a stable capacity of 41 mAh/g at 2C rate for 430 cycles (calculated for the weight of NMO cathode). Even at a fast 5C rate, a discharge capacity of 33 mAh/g was maintained for 2,400 prolonged cycles with nearly 100% efficiency. The full cell device can achieve an average voltage of 1 V with energy density of 24 Wh/kg. This study highlights concentrated sodium perchlorate as a promising electrolyte solution for stabilization of electrodes and enhancement of electrochemical performance in aqueous media.

Survival and growth of soil microbial communities under influence of sodium perchlorates

AbstractPreviously conducted space missions revealed the presence of perchlorates, which are known to have a high oxidizing potential in Martian regolith, at the level of 0.5%. Due to hygroscopic properties and crystallization features of perchlorate-containing solutions, assumptions leading to the possibility of the existence of liquid water in the form of brines, which can contribute to the vital activity of microorganisms, have been made. At the same time, high concentrations of perchlorates can inhibit the growth of microorganisms and cause their death. Previously performed studies have discovered the presence of highly diverse microbial communities in terrestrial perchlorate-containing soils and have also demonstrated the stability and activity of some prokaryotes cultured on highly concentrated perchlorates media (over 10%). Nevertheless, the limits of microbial tolerance to perchlorates and whether microbial communities are able to withstand the effects of high concentrations of perchlorates remain uncertain. The aim of this research was to study the reaction of microbial communities of hot-arid and cryo-arid soils and sedimentary rocks to the adding of a highly concentrated solution of sodium perchlorate (5%) in situ. An increase in the total number of prokaryotes, the number of metabolically active Bacteria and Archaea, and the variety of the consumed substrates were revealed. It was observed that in samples incubated with sodium perchlorate, a high taxonomic diversity of the microbial community is preserved at a level comparable to control sample. The study shows that the presence of high concentrations of sodium perchlorate (5%) in the soil does not lead to the death or significant inhibition of microbial communities.

Separation and pre-concentration of Sunset Yellow in beverages and effervescent vitamin C tablets by a new flotation technique prior to spectrophotometric determination

ABSTRACTA new, simple, low cost, high efficiency, and rapid floating technique was developed for the determination of Sunset Yellow in beverage and effervescent vitamin C. The method is based on two main steps: 1)- formation of dye-surfactant complex by the addition of a cationic surfactant, cetyltrimethylammonium bromide, to the solution containing an anionic dye (Sunset Yellow) and 2)- flocculation of the dye-surfactant complex with the participation of sodium perchlorate as a coagulant followed by aggregation and enlargement of the flocs while floating at the top of the solution. After the extraction, separated and preconcentrated Sunset Yellow was investigated by spectrophotometric determination. IR spectrum, scanning electron microscopy (SEM), and light microscopy were used to characterise the flocs. The effects of different parameters such as pH, concentration of surfactant, concentration of sodium perchlorate, and temperature on the extraction of dye were investigated and optimized. Under optimum conditions, a linear calibration curve was obtained in the range of 0.05–5 mg L−1. The detection limit (3Sb/m) was 0.02 mg L−1 with a preconcentration factor of 25. The relative standard deviation (RSD) for 0.1 mg L−1 of Sunset Yellow was 2.6% (n = 10). The applicability of the method was evaluated by measuring the amount of dye in beverage and effervescent vitamin C. Interferences of typical cations and anions which may participate in the matrices were also checked. The amount of Sunset Yellow found in real samples was 1.32–3.89 mgL−1.

High Tolerance of Hydrogenothermus marinus to Sodium Perchlorate.

On Mars, significant amounts (0.4–0.6%) of perchlorate ions were detected in dry soil by the Phoenix Wet Chemistry Laboratory and later confirmed with the Mars Science Laboratory. Therefore, the ability of Hydrogenothermus marinus, a desiccation tolerant bacterium, to survive and grow in the presence of perchlorates was determined. Results indicated that H. marinus was able to tolerate concentrations of sodium perchlorate up to 200 mM ( 1.6%) during cultivation without any changes in its growth pattern. After the addition of up to 440 mM ( 3.7%) sodium perchlorate, H. marinus showed significant changes in cell morphology; from single motile short rods to long cell chains up to 80 cells. Furthermore, it was shown that the known desiccation tolerance of H. marinus is highly influenced by a pre-treatment with different perchlorates; additive effects of desiccation and perchlorate treatments are visible in a reduced survival rate. These data demonstrate that thermophiles, especially H. marinus, have so far, unknown high tolerances against cell damaging treatments and may serve as model organisms for future space experiments.

Thermal, aggregation, counterion binding, light scattering, and adsorption behavior of cis-chlorobis(ethylenediamine)dodecylaminecobalt(III) perchlorate metallosurfactant in aqueous sodium perchlorate medium

Studying the fundamental aspects of aggregation and adsorption of metallosurfactants is necessary by looking at the potential applications of these novel surfactants. cis-Chlorobis(ethylenediamine)dodecylaminecobalt(III) perchlorate (CDCP) metallosurfactant was synthesized and its thermal gravimetric analysis showed that it is stable up to 217 °C. Critical micelle concentration (cmc) of CDCP in aqueous sodium perchlorate medium was determined by using surface tension, conductivity, and spectrophotometric methods. It is shown that the cmcs of metallosurfactants do not follow the trend according to the positions of the counterions in the Hofmeister series, which is contrary to the trend observed in the case of conventional ionic surfactants. The counterion binding constant of CDCP is equal to 0.81. The variation of surface excess of CDCP with sodium perchlorate concentration exhibits a minimum. Dynamic light scattering measurements showed the presence of large size aggregates of CDCP in solution, and these aggregates grow further as the counterion concentration increased.

Retention Behavior of Inorganic Anions in Hydrophilic Interaction Chromatography.

The retention behavior of inorganic anions was studied in hydrophilic interaction chromatography (HILIC). In this study, five kinds of HILIC stationary phases (amino, imidazole, amide, pyridine and zwitterionic) were investigated. It was found that only amino and imidazole columns exhibited the separation of inorganic anions under HILIC conditions. The retention mechanism was further investigated under both columns. A reversed elution order of inorganic anions was observed under the HILIC condition compared with those observed under the ion-exchange chromatography mode (IEC). The effect of salt species and their concentration in the eluent were investigated under constant acetonitrile (ACN) content. Sodium chloride and sodium perchlorate were chosen as the salt, and the salt (sodium perchlorate) concentration was varied from 10 to 40 mM to confirm the effect of the electrostatic interaction. The slope values of the plots of the log retention factor (k) versus the log eluent concentration were calculated to be between -0.43 and -0.45 for the amino column, while those obtained on the imidazole column were between -0.68 and -0.73. Various concentrations of ACN were also examined with 20 mM sodium perchlorate, and the typical HILIC retention behavior was observed on both amino and imidazole columns. Due to the obtained results, it is considered that the separation of inorganic anions under the HILIC condition was achieved by both electrostatic interaction and partition.

Synthesis of Zn:Cu2O thin films using a single step electrodeposition for photovoltaic applications.

Zinc-doped cuprous oxide (Zn:Cu2O) thin films have been prepared via single step electrodeposition from an aqueous solution containing sodium perchlorate. The Zn/Cu molar ratio in the Zn:Cu2O films can be tuned between 0.006 and 0.236 by adjusting the magnitude of the applied potential and the sodium perchlorate concentration. Electrical characterization reveals that zinc dopants increase the Fermi level in Zn:Cu2O films, enabling a 3-fold improvement in the power conversion efficiency of a fully electrodeposited Cu2O homojunction photovoltaic device.

Effects of water temperature on perchlorate toxicity to the thyroid and reproductive system of Oryzias latipes

Water temperature is expected to increase in many parts of the world due to global climate change. The change in water temperature may affect ecosystems through alterations of the chemical properties or by affecting the susceptibility of organisms. Perchlorate can disrupt thyroid function of an organism by inhibiting iodide uptake. In the present study, the effect of water temperature on perchlorate toxicity was evaluated using Japanese medaka (Oryzias latipes). Pairs of adult medaka fish were exposed to a sublethal concentration of sodium perchlorate (100mg/L) and a control, at a ‘low’ (26°C), ‘medium’ (29°C) or ‘high’ water temperature (33°C) for seven days. The effects of the water temperature on reproduction, thyroid hormones and cortisol concentrations were determined. Transcription of several genes related to thyroid function and stress were also investigated. Significant down-regulation of thyroid hormone receptor alpha (THR-α) and beta (THR-β) transcripts and up-regulation of deiodinase 2 (DIO2) transcripts were observed in the fish exposed to perchlorate. Thyroxine (T4) concentrations were decreased, while triiodothyronine (T3) levels remained constant following exposure to perchlorate, and this effect became more pronounced under the high water temperature conditions (33°C). Up-regulation of the DIO2 gene may explain these observations. The total number of spawned eggs decreased slightly as the water temperature increased, and this reduction became significant when fish were exposed to perchlorate. Our observations indicate that exposure to perchlorate could affect thyroid function and overall reproductive fitness, and these effects could be aggravated under high water temperatures.

Theranostic studies of human sodium iodide symporter imaging and therapy using 188Re: a human glioma study in mice.

ObjectiveTo investigate the role of 188Re in human sodium iodide symporter (hNIS) theranostic gene-mediated human glioma imaging and therapy in model mice.MethodsThe human glioma cell line U87 was transfected with recombinant lentivirus encoding the hNIS gene under the control of cytomegalovirus promoter (U87-hNIS). The uptake and efflux of 188Re were determined after incubating the cells with 188Re. 188Re uptake experiments in the presence of various concentrations of sodium perchlorate were carried out. In vitro cell killing tests with 188Re were performed. U87-hNIS mediated 188Re distribution, imaging and therapy in nude mice were also tested.ResultsU87-hNIS cell line was successfully established. The uptake of 188Re in U87-hNIS cells increased up to 26-fold compared to control cells, but was released rapidly with a half-life of approximately 4 minutes. Sodium perchlorate reduced hNIS-mediated 188Re uptake to levels of control cell lines. U87-hNIS cells were selectively killed following exposure to 188Re, with a survival of 21.4%, while control cells had a survival of 92.1%. Unlike in vitro studies, U87-hNIS tumor showed a markedly increased 188Re retention even 48 hours after 188Re injection. In the therapy study, there was a significant difference in tumor size between U87-hNIS mice (317±67 mm3) and control mice (861±153 mm3) treated with 188Re for 4 weeks (P<0.01).ConclusionThe results indicate that inserting the hNIS gene into U87 cells is sufficient to induce specific 188Re uptake, which has a cell killing effect both in vitro and in vivo. Moreover, our study, based on the function of hNIS as a theranostic gene allowing noninvasive imaging of hNIS expression by 188Re scintigraphy, provides detailed characterization of in vivo vector biodistribution and level, localization, essential prerequisites for precise planning and monitoring of clinical gene therapy that aims to individualize gene therapy concept.

Hydrogen generation by tin corrosion in lactic acid solution promoted by sodium perchlorate

A method to produce high purity hydrogen using the corrosion of tin metal in lactic acid solutions is studied. The addition of sodium perchlorate has been also investigated for promoting the tin-lactic acid reaction. The data reveal that the rate of hydrogen production increases with increasing lactic acid concentration. The presence of perchlorate ions in lactic acid solution enhances the active dissolution of tin metal and tends to breakdown the passive film and promoting the hydrogen generation rate. Polarization measurements show that the breakdown potential (Epit) decreases with increase in sodium perchlorate concentration. An increase in temperature accelerates the rate of solubility of passive layer on the tin surface. Moreover, a synergistic effect of sodium perchlorate in combination with increasing the solution temperature is key in promoting the hydrogen generation rate. Results obtained from hydrogen and polarization measurements are in good agreement. These measurements are complemented with SEM, EDX and XRD examinations of the electrode surface.

Optimization of ion‐pair based hollow fiber liquid phase microextraction combined with HPLC–UV for the determination of methimazole in biological samples and animal feed

Ion-pair based hollow fiber liquid phase microextraction (IP-HFLPME) coupled with high performance liquid chromatography-ultraviolet detection was applied for the preconcentration and determination of methimazole in biological samples and animal feed. Optimization of the conditions for the high extraction efficiency was studied simultaneously using the experimental design. For the first step, the Plackett-Burman design was applied to screen the significant factors on the extraction efficiency. Central composite design (CCD) was then used for the optimization of important factors and the response surface equations were obtained. The optimum experimental conditions were donor phase pH, 12.2; extraction temperature, 45°C; extraction time, 50 min; sodium perchlorate concentration, 1.5 M; cetyltrimethylammonium bromide concentration, 0.65 mM, and without salt addition in donor phase. The limit of detection and the dynamic linear range were in the range of 0.1-0.7 μg L(-1) and 0.5-1000 μg L(-1), respectively. Preconcentration factors were obtained in the range of 93-155 in different matrices. Finally, the performance of the proposed method was tested for the determination of trace amounts of methimazole in plasma, urine, bovine milk, and animal feed samples, and satisfactory results were obtained (RSDs < 7.1%).

Effect of sodium and acetate ions on 8-hydroxyguanine formation in irradiated aqueous solutions of DNA and 2′-deoxyguanosine 5′-monophosphate

Purpose: The aim of this work was to study the combined effect of sodium and acetate ions on the radiation yield of 8-hydroxyguanine (8-OHG), one of the major DNA base lesions induced by free radicals.Materials and methods: Aqueous solutions of DNA and 2′-deoxyguanosine 5′-monophosphate (dGMP) with various concentrations of sodium acetate and sodium perchlorate were γ-irradiated, enzymatically digested and analyzed by high-performance liquid chromatography (HPLC) methods.Results: It was found that both salts decrease the 8-OHG radiation yield in the concentration range studied for both DNA and dGMP, except in the case of dGMP wherein an increase in yield occurs in the concentration range from 0.1–1 mM. The dependence of the 8-hydroxy-2′-deoxyguanosine radiation yield on the concentration of both sodium acetate and sodium perchlorate have different shapes and have steeper slopes for the DNA compared with the dGMP solutions.Conclusions: The observed decrease in the radiation yield of 8-OHG with increasing concentrations of sodium acetate is consistent with the hypothesis that sodium acetate produces two concentration-dependent effects in the DNA solutions: (1) A conformational change in the DNA caused by Na+ counterions; and (2) free radical reactions related to the radiolysis of acetate ion.

Influence of Water Temperature on Perchlorate-induced Toxicity in Oryzias latipes

PP-29-086 Background/Aims: Thyroid hormones play an essential role in the metabolism, growth, and development in vertebrate. Some synthetic chemicals such as perchlorate have been reported to disrupt thyroid function. Perchlorate interferes with iodine uptake in the thyroid hormone synthesis process. Water temperature can induce stress hormone cortisol, which may influence thyroid function as well. However, the effect of water temperature on perchlorate-induced thyroid toxicity is not well understood. Methods: In this study, Oryzias latipes were exposed to sublethal concentration (100 mg/L) of sodium perchlorate at 3 different water temperature, that is, low (26°C), medium (29°C), and high temperature (33°C) for 7 days, and the effect of water temperature on perchlorate toxicity was observed. Results: Perchlorate exposure decreased transcription of both thyroid hormone receptor alpha and beta mRNAs. Perchlorate exposure at high water temperature induced upregulations of mRNA levels of both NaKATPase alpha and beta, presumably because of increased induction of stress hormone cortisol. Moreover, as water temperature increased, fecundity of fish, eg, number of eggs per pair was decreased. Conclusion: The results of our study suggested that the decrease of reproduction might be partly explained by disruption in thyroid function, which was resulted from inhibition of thyroid hormone synthesis by perchlorate exposure and interference of tissue response to thyroid hormone signal by high water temperature.

ChemInform Abstract: Evaluation of the Activity Coefficients of Some Sulfonated Azo Dyes and of Their Complexes with Metal Ions

The protonation and complexation conditional constants of equilibria involving large, multicharged organic ions depend on the ionic medium in which they occur. The possibility of evaluating these effects on the basis of the Specific Interaction Theory (SIT) was tested, by determining spectrophotometrically the conditional protonation constants (in concentration) of some bisulfonated azo dyes [2-hydroxy-1-(1H-tetrazol-5-ylazo)naphthalene-3,6-disulfonic acid, 1,8-dihydroxy-2-(1H-tetrazol-5-ylazo)naphthalene-3,6-disulfonic acid and 1,8-dihydroxy-2-(p-nitrophenylazo)-naphthalene-3,6-disulfonic acid] at different sodium perchlorate concentrations. The same was done for complexation with thorium(IV), indium(III) and copper(II). It was found that the dependance of protonation and complexation conditional constants on the ionic medium can be described by evaluating the activity coefficients of the ions on the basis of SIT. The activity coefficients of the multicharged free organic ligands are best estimated by considering their charge globally, exactly like in the case of small inorganic ions. On the other hand, the activity coefficients of the complexes with a metal ion are best evaluated by considering them as bearing two separate charged sites, one corresponding to the site where the metal ion is bound, and the other to the residual charge of the ligand. In this way the specific interaction coefficients were found to be constant, as expected from SIT, and their summations were experimentally determined, together with the thermodynamic constants, by linear extrapolation.

Analysis of cholesterol levels in lipoprotein(a) with anion-exchange chromatography

We previously established an analysis method for determining the cholesterol levels of five major lipoprotein classes [HDL, LDL, intermediate density lipoprotein (IDL), VLDL, and chylomicrons] in serum by an anion-exchange (AEX)-HPLC method, but lipoprotein(a) [Lp(a)], a well-known risk factor for atherosclerotic diseases, was not determinable. Therefore, we established new AEX-HPLC separation conditions for analyzing the cholesterol levels of six lipoprotein classes, including Lp(a). Serum lipoproteins were separated by HPLC with a diethylaminoethyl-ligand nonporous polymer-based column by elution with a stepwise gradient of the sodium perchlorate concentration. In this improved method, HDL, LDL, IDL, VLDL, chylomicrons, and Lp(a) were each eluted from the column. The cholesterol levels of the eluted lipoproteins were measured enzymatically by a postcolumn reaction. The within-day assay and between-day assay coefficients of variation for the lipoprotein cholesterol levels were in the ranges of 0.29-11.86% and 0.57-11.99%, respectively. The Lp(a) cholesterol levels determined by AEX-HPLC were significantly correlated with the amounts of Lp(a) protein measured by an immunoturbidimetry method available commercially (r = 0.9503, P < 0.0001). Taken together, this AEX-HPLC method may be effectively applied to the analysis of serum lipoproteins with high levels of Lp(a).