Splicing factor ESRP1 derived circ_0068162 promotes the progression of oral squamous cell carcinoma via the miR-186/JAG axis.

OBJECTIVES: Oral squamous cell carcinoma (OSCC) is a common malignancy in the oral and maxillofacial regions with an increasing incidence rate. Circular RNA (circRNA) is a recently discovered long-chain non-coding RNA family member. The objective of this study was to analyze the role of circ_0068162 in OSCC development. METHODS: We downloaded sample data GSE145608 from the Gene Expression Omnibus database. Online databases Starbase, TargetScan and miRDB were used to predict the target microRNAs (miRNAs) and genes. Cell viability and proliferation were assessed using the CCK-8 and EdU assays, respectively. Cell migration and invasion abilities were detected using transwell assay. The double luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to verify the interaction relationship between the identified target molecules. RNase R and actinomycin D treatment were performed to analyze the stability of circ_0068162. RESULTS: We found that circ_0068162 was overexpressed in the cytoplasm of OSCC cells and clinical OSCC tissues. Knockdown of circ_0068162 inhibited the growth, migration and invasion of OSCC cells. We also identified miR-186 as the target miRNA of circ_0068162, and JAG1 and JAG2 as the target genes of miR-186. The miR-186 inhibitor rescued the effects of sh-circ_0068162 and JAG1/JAG2 overexpression rescued the effects of miR-186 mimic in OSCC cells. Furthermore, ESRP1 promoted the biosynthesis of circ_0068162. CONCLUSIONS: The circ_0068162/miR-186/JAGs/ESRP1 feedback loop is closely related to OSCC development.

Magnetic porous aromatic framework with a core-shell structure as a sorbent for rapid extraction of phenols and their quantitation in urine by HPLC-UV.

A porous aromatic framework (PAF) is shown to be a viable sorbent for the adsorption of phenols. To overcome the difficulty of quick adsorption and enrichment by phenols from the matrix, a sorbent material consisting of porous aromatic framework magnetic nanoparticles (PAF MNPs) with a core-shell structure was fabricated by an in situ growth method. The PAF MNP sorbent was characterized by transmission electron microscopy, Fourier transform infrared spectroscopy and other techniques. The factors affecting enrichment performance including the amount of PAF-6 MNPs, sample pH, extraction time and elution conditions were optimized. Under the optimal conditions, a method utilizing high-performance liquid chromatography with an ultraviolet detector (HPLC-UV) was developed to quantify phenols in urine. The method showed good linearity (r ≥ 0.998), good precision (RSD ≤ 9.9%, n = 6) and a low limit of detection (1.0-2.0 ng/mL, S/N = 3), with recoveries performed in urine matrix ranging from 76.7% to 113.2%. The method is simple, time-saving and sensitive. Moreover, compared with traditional mass spectrometry detection methods, this method has advantages in terms of low cost and repeatability. Graphical abstract.

Correlation of urine protein/creatinine ratios to 24-h urinary protein for quantitating proteinuria in children.

BACKGROUND: The purpose of this study was to evaluate the correlation and consistency between urine protein/creatinine ratio (UPCR) and 24-h urine protein (24HUPr) in children, and to determine cutoff values of UPCR relative to 24HUPr at 100 mg/m2/d (≥ 100 mg/m2/d as pathological proteinuria) and 1000 mg/m2/d (≥ 1000 mg/m2/d as nephrotic-range proteinuria). METHODS: Three hundred sixty-six children were enrolled, including 81 controls, 109 with Henoch-Schönlein purpura nephritis, 167 with nephrotic syndrome, 5 with IgA nephropathy, and 4 with lupus nephritis. Patients were divided into three groups: normal group; non-nephrotic-range proteinuria group; nephrotic-range proteinuria group. The cutoff values of UPCR in predicting the different levels of proteinuria were determined using ROC curve analysis. RESULTS: UPCR was positively correlated with 24HUPr (r = 0.915, p < 0.01). Bland-Altman diagrams showed that UPCR and 24HUPr had good consistency, and > 95% spots of UPCR and 24HUPr were within 95% confidence intervals. Relative to 24HUPr at 100 mg/m2/d, the cutoff value of UPCR (0.18 g/g Cr) had the highest sensitivity (94%) and specificity (98.8%) which is close to 0.2 g/g Cr as proposed by the American College of Rheumatology. Relative to 24HUPr at 1000 mg/m2/d, the cutoff value of UPCR (2.09 g/g Cr) had the highest sensitivity (92.1%) and specificity (92.1%) which is close to the 2.0 g/g Cr proposed in KDIGO guidelines. CONCLUSIONS: UPCR showed strong correlation and consistency with 24HUPr for evaluating levels of proteinuria in children. UPCR < 0.2 g/g Cr can be considered a criterion for normal-range proteinuria. Instead of 24HUPr ≥ 1000 mg/m2/d, UPCR ≥ 2.0 g/g Cr can be considered a criterion for nephrotic-range proteinuria or nephrotic syndrome in children.

Development of a decaaza-cyclophane stationary phase for high-performance liquid chromatography.

A new stationary phase for high-performance liquid chromatography was prepared by covalently bonding a heteroatom-bridged cyclophane onto silica gel using 3-aminopropyltriethoxysilane as the coupling reagent. The structure of the new material was characterized by infrared spectroscopy, elemental analysis, and thermogravimetric analysis. The linear solvation energy relationship method was successfully employed to evaluate the new phase with a set of 25 solutes, and compared with octadecylsilyl and p-tert-butyl-calix[4]arene bonded stationary phases. The retention characteristics of the new phase are similar to the octadecylsilyl and conventional calixarene phases, and it also has distinctive features. In addition, the chromatographic behavior of the phase was illustrated by eluting alkylbenzenes and inorganic anions in the reversed-phase mode and anion-exchange mode, respectively. Thus, multi-interaction mechanisms and mixed-mode separation of the new phase can very likely guarantee its promising application in the analysis of complex samples. The column has been successfully employed for the analysis of triazines in milk, and it is demonstrated to be a competitive alternative analytical method for the determination of triazine herbicide residues.

Liquid chromatographic behavior of two alanine-substituted calix[4]arene-bonded silica gel stationary phases.

Two new kinds of alanine-substituted calix[4]arene stationary phases of 5,11,17,23-p-tert-butyl-25,27-bis(l-alanine-methylester-N-carbonyl-methoxy)-26,28-dihyroxycalix[4]arene-bonded silica gel stationary phase (BABS4) and 5, 11, 17, 23-p-tert-butyl-25,26,27,28-tetra(l-alanine-methylester-N-carbonyl-methoxy)-calix[4]arene-bonded silica gel stationary phase (TABS4) were prepared and characterized in the present study. They were compared with each other and investigated in terms of their chromatographic performance by using polycyclic aromatic hydrocarbons, disubstituted benzene isomers, and mono-substituted benzenes as solute probes. The results indicated that both BABS4 and TABS4 exhibited multiple interactions with analytes. In addition, the commonly used Tanaka characterization protocol for the evaluation of commercially available stationary phases was applied to evaluate the properties of these two new functionalized calixarene stationary phases. The Tanaka test results were compared with Zorbax Eclipse XDB C18 and Kromasil phenyl columns, respectively. BABS4 has stronger hydrogen-bonding capacity and ion-exchange capacity than TABS4, and features weaker hydrophobicity and hydrophobic selectivity. Both of them behave similarly in stereoselectivity. Both BABS4 and TABS4 are weaker than C18 and phenyl stationary phases in hydrophobicity and hydrophobic selectivity.

Linear ubiquitination regulates the KSHV replication and transcription activator protein to control infection.

Like many other viruses, KSHV has two life cycle modes: the latent phase and the lytic phase. The RTA protein from KSHV is essential for lytic reactivation, but how this protein's activity is regulated is not fully understood. Here, we report that linear ubiquitination regulates the activity of RTA during KSHV lytic reactivation and de novo infection. Overexpressing OTULIN inhibits KSHV lytic reactivation, whereas knocking down OTULIN or overexpressing HOIP enhances it. Intriguingly, we found that RTA is linearly polyubiquitinated by HOIP at K516 and K518, and these modifications control the RTA's nuclear localization. OTULIN removes linear polyubiquitin chains from cytoplasmic RTA, preventing its nuclear import. The RTA orthologs encoded by the EB and MHV68 viruses are also linearly polyubiquitinated and regulated by OTULIN. Our study establishes that linear polyubiquitination plays a critically regulatory role in herpesvirus infection, adding virus infection to the list of biological processes known to be controlled by linear polyubiquitination.

The mechanism of epigallocatechin-3-gallate inhibiting the antigenicity of ß-lactoglobulin under pH 6.2, 7.4 and 8.2: Multi-spectroscopy and molecular simulation methods.

The antigenicity of ß-lactoglobulin (ß-LG) can be influenced by pH values and reduced by epigallocatechin-3-gallate (EGCG). However, a detailed mechanism concerning EGCG decreasing the antigenicity of ß-LG at different pH levels lacks clarity. Here, we explore the inhibition mechanism of EGCG on the antigenicity of ß-LG at pH 6.2, 7.4 and 8.2 using enzyme-linked immunosorbent assay, multi-spectroscopy, mass spectrometry and molecular simulations. The results of Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD) elucidate that the noncovalent binding of EGCG with ß-LG induces variations in the secondary structure and conformations of ß-LG. Moreover, EGCG inhibits the antigenicity of ß-LG the most at pH 7.4 (98.30 %), followed by pH 6.2 (73.18 %) and pH 8.2 (36.24 %). The inhibitory difference is attributed to the disparity in the number of epitopes involved in the interacting regions of EGCG and ß-LG. Our findings suggest that manipulating pH conditions may enhance the effectiveness of antigenic inhibitors, with the potential for further application in the food industry.

CETSA-MS-based target profiling of anti-aging natural compound quercetin.

BACKGROUND: Quercetin is widely distributed in nature and abundant in the human diet, which exhibits diverse biological activities and potential medical benefits. However, there remains a lack of comprehensive understanding about its cellular targets, impeding its in-depth mechanistic studies and clinical applications. PURPOSE: This study aimed to profile protein targets of quercetin at the proteome level. METHODS: A label-free CETSA-MS proteomics technique was employed for target enrichment and identification. The R package Inflect was used for melting curve fitting and target selection. D3Pocket and LiBiSco tools were used for binding pocket prediction and binding pocket analysis. Western blotting, molecular docking, site-directed mutagenesis and pull-down assays were used for target verification and validation. RESULTS: We curated a library of direct binding targets of quercetin in cells. This library comprises 37 proteins that show increased thermal stability upon quercetin binding and 33 proteins that display decreased thermal stability. Through Western blotting, molecular docking, site-directed mutagenesis and pull-down assays, we validated CBR1 and GSK3A from the stabilized protein group and MAPK1 from the destabilized group as direct binding targets of quercetin. Moreover, we characterized the shared chemical properties of the binding pockets of quercetin with targets. CONCLUSION: Our findings deepen our understanding of the proteins pivotal to the bioactivity of quercetin and lay the groundwork for further exploration into its mechanisms of action and potential clinical applications.

Reducing Antigenicity and Improving Antioxidant Capacity of ß-Lactoglobulin through Covalent Interaction with Six Flavonoids.

ß-lactoglobulin (ß-LG) is a pivotal nutritional and functional protein. However, its application is limited by its antigenicity and susceptibility to oxidation. Here, we explore the impact of covalent modification by six natural compounds on the antigenicity and antioxidant characteristics of ß-LG to explore the underlying interaction mechanism. Our findings reveal that the covalent interaction of ß-LG and flavonoids reduces the antigenicity of ß-LG, with the following inhibition rates: epigallocatechin-3-gallate (EGCG) (57.0%), kaempferol (42.4%), myricetin (33.7%), phloretin (28.6%), naringenin (26.7%), and quercetin (24.3%). Additionally, the ß-LG-flavonoid conjugates exhibited superior antioxidant capacity compared to natural ß-LG. Our results demonstrate that the significant structural modifications from α-helix to ß-sheet induced by flavonoid conjugation elicited distinct variations in the antigenicity and antioxidant activity of ß-LG. Therefore, the conjugation of ß-LG with flavonoids presents a prospective method to reduce the antigenicity and enhance the antioxidant capacity of ß-LG.

Insight into covalent conjugates of ß-lactoglobulin with rutin: Characterizing allergenicity, digestibility, and antioxidant properties in vitro.

ß-lactoglobulin (ß-LG) is an essential nutrient in milk, but it is the primary allergen causing dairy allergy in humans. Currently, researchers are focusing on using flavonoids to covalently modify ß-LG for improving its functionality. However, the impact and underlying mechanisms of rutin covalent modification on the functional properties and allergenicity of ß-LG remain unclear. Here, we aim to investigate the changes in allergenicity, digestive characteristics, and antioxidant properties of ß-LG after covalent modification using a combination of spectroscopy, enzyme-linked immunosorbent assay (ELISA), simulated digestion, and antioxidant assays. The results indicate that rutin forms covalent bonds with the free amino group, sulfhydryl group, and tryptophan of ß-LG, leading to alterations in the secondary structure of ß-LG. Furthermore, the modified ß-LG exhibits improved antioxidant capacity and decreased allergenicity, along with reduced resistance to pancreatin digestion in vitro. This study provides novel insights and strategies to expand the functional application of ß-LG.

Insight into the Interaction Mechanism of Vitamin D against Metabolic Syndrome: A Meta-Analysis and In Silico Study.

As a dietary supplement or functional food additive, vitamin D (VD) deficiency may impact extra-skeletal functions associated with metabolic syndrome (MetS) risk factors. However, the precise effects and mechanisms of VD supplementation on dyslipidemia and insulin resistance in MetS subjects remain controversial. Here, we investigate potential therapeutic targets, pathways and mechanisms of VD against MetS through a comprehensive strategy including meta-analysis, network pharmacology analysis, molecular docking, dynamics simulations, and quantum chemical calculations. Our results reveal that VD supplementation significantly reduces triglyceride levels, fasting glucose, and insulin concentrations in subjects, thereby improving insulin homeostasis to some extent. We theoretically identify 14 core MetS-associated targets. Notably, VD exhibits substantial interactions with three targets (PPARγ, FABP4, and HMGCR) in the PPAR signaling pathway, indicating that VD can modulate this pathway. Van der Waals forces predominantly stabilize the complexes formed between VD and the three targets. Nonetheless, to provide valuable insights for personalized MetS management, further research is necessary to confirm our findings, emphasizing the importance of exploring genetic variability in VD response. In conclusion, our study contributes insights into the mechanisms of VD in preventing and treating MetS through dietary supplementation, promoting the development of VD-based functional foods or nutritious diets.

Strategy to combat biofilms: a focus on biofilm dispersal enzymes.

Bacterial biofilms, which consist of three-dimensional extracellular polymeric substance (EPS), not only function as signaling networks, provide nutritional support, and facilitate surface adhesion, but also serve as a protective shield for the residing bacterial inhabitants against external stress, such as antibiotics, antimicrobials, and host immune responses. Biofilm-associated infections account for 65-80% of all human microbial infections that lead to serious mortality and morbidity. Tremendous effort has been spent to address the problem by developing biofilm-dispersing agents to discharge colonized microbial cells to a more vulnerable planktonic state. Here, we discuss the recent progress of enzymatic eradicating strategies against medical biofilms, with a focus on dispersal mechanisms. Particularly, we review three enzyme classes that have been extensively investigated, namely glycoside hydrolases, proteases, and deoxyribonucleases.

Shared metabolic shifts in endothelial cells in stroke and Alzheimer's disease revealed by integrated analysis.

Since metabolic dysregulation is a hallmark of both stroke and Alzheimer's disease (AD), mining shared metabolic patterns in these diseases will help to identify their possible pathogenic mechanisms and potential intervention targets. However, a systematic integration analysis of the metabolic networks of the these diseases is still lacking. In this study, we integrated single-cell RNA sequencing datasets of ischemic stroke (IS), hemorrhagic stroke (HS) and AD models to construct metabolic flux profiles at the single-cell level. We discovered that the three disorders cause shared metabolic shifts in endothelial cells. These altered metabolic modules were mainly enriched in the transporter-related pathways and were predicted to potentially lead to a decrease in metabolites such as pyruvate and fumarate. We further found that Lef1, Elk3 and Fosl1 may be upstream transcriptional regulators causing metabolic shifts and may be possible targets for interventions that halt the course of neurodegeneration.

Comprehensive profiling of Stephania tetrandra (Fangji) by stepwise DFI and NL-dependent structure annotation algorithm-based UHPLC-Q-TOF-MS and direct authentication by LMJ-HRMS.

Stephania tetrandra S. Moore, a widely used traditional antirheumatic herbal medicine (HM), is a rich source of isoquinoline alkaloids. With the exception of the two recognized isoquinolines, viz. tetrandrine and fangchinoline, the other isoquinoline alkaloids present in S. tetrandra have not been clearly clarified. In addition, due to their similar names and morphological similarities, S. tetrandra is often mistakenly substituted and adulterated with the nephrotoxic Aristolochia fangchi. In this study, ultra-high-performance liquid chromatography-triple time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was initially employed to comprehensively profile the isoquinolines from S. tetrandra. To overcome the complexities arising due to the similar mass behaviors of the isoquinolines, a stepwise diagnostic fragment ion (DFI) and neutral loss (NL)-dependent structure annotation algorithm was proposed, and this accelerated the identification of 393 isoquinolines distributed over twenty classes. Consequently, liquid microjunction surface sampling-high-resolution mass spectrometry (LMJ-HRMS) was deployed in an attempt to directly authenticate S. tetrandra by the chemical profiling of its crude slice. By matching the 393 isoquinolines, the 87 peaks detected by LMJ-HRMS were assigned to 270 isoquinolines, including the recognized tetrandrine and fangchinoline. The absence of aristolochic acid-related mass signals confirmed the authentication of S. tetrandra. In summary, LMJ-HRMS can be considered a direct, nondestructive, high-throughput, and environment-friendly analytical method for the authentication of HMs. Moreover, the stepwise DFI- and NL-dependent structure annotation algorithm-based UHPLC-Q-TOF-MS method allowed high-coverage detection and high-quality data processing of the inherent structural similarity and complexity of isoquinolines or other phytochemical compounds.

Tetra-proline modified calix[4]arene bonded silica gel: A novel stationary phase for hydrophilic interaction liquid chromatography.

Calixarenes and their derivatives have been widely used in chromatographic studies due to unique host-guest recognition properties. However, stationary phases in hydrophilic interaction chromatography associated with them have yet to be studied because of their general hydrophobicity. In this paper, a tetra-proline modified calix[4]arene bonded stationary phase (DTPCSP) was prepared and characterized by FT-IR spectra, elemental analysis, solid state 13C NMR, SEM, EDS and thermogravimetric analysis. The chromatographic performance and retention mechanism of the developed stationary phase were validated in hydrophilic interaction mode and compared with a commercial column using a variety of hydrophilic or hydrophobic compounds including phenols, nucleosides and sulfonamides. The developed stationary phase exhibited better selectivity than conventional calixarene phases, and the retention behaviors of phenols on DTPCSP column were elucidated by the calculation of quantum chemistry using ODS as contrast. All the results indicate that the developed DTPCSP stationary phase can be beneficial for simultaneously separating complex hydrophilic samples with high selectivity.

Determination of benzo[α]pyrene in edible oil using tetraoxocalix[2]arene[2]triazine bonded silica SPE sorbent.

Benzo[α]pyrene (BaP) is a well-known carcinogen in edible oil. In this study, a method combined solid-phase extraction (SPE) with fluorescent detection was developed using tetraoxocalix[2]arene[2]triazine sorbent (SiO2-OCA) for the clean-up and enrichment of BaP. The interaction between SiO2-OCA and BaP involves a donor-acceptor complex mechanism. The experimental procedure was as follows: BaP was extracted from edible oil with DMF/H2O (9:1, v/v). Then, the ratio of DMF/H2O was adjusted to 1:2 prior to SPE. The final concentrate was analysed using a fluorescence detector at excitation and emission wavelengths of 255 and 420 nm. The method was fully validated. The linearity was in the range of 0.1-100 µg kg-1 with a coefficient of 0.999. The limits of detection and quantification were 0.03 and 0.1 µg kg-1, respectively. The average recoveries were in the range of 88.0 - 122.3%. The intraday and interday precisions were 6.8% and 9.2%, respectively. Compared with other methods, the method reported in this article shows a good detection limit, high reproducibility and recovery and linearity over a broad concentration range. This established method was also applied to evaluate real samples. The concentration of six tested samples was below 5 µg kg-1.

In situ preparation of core-shell magnetic porous aromatic framework nanoparticles for mixed-mode solid-phase extraction of trace multitarget analytes.

This study reports a novel strategy for the preparation of porous aromatic framework (PAF-6) coated magnetic nanoparticles (PAF-6 MNPs) using cyanuric chloride as a planar trigonal basis upon which to build a linear piperazine linker unit. The PAF-6 MNPs were examined as an efficient solid-phase extraction (SPE) sorbent for enrichment of trace organic pollutants including phenol, 2,4,6-trinitrophenol, naphthalene, naphthol, bisphenol A, 2,4-dichlorophenol and 3-nitrochlorobenzene in water. The high-performance liquid chromatography detection limits of such analytes were in the range of 0.08-5.02 ng/mL and recoveries were found to be 84.0-94.0% in well water, tap water, river water and wastewater. The main toxic components of cigarette smoke, including phenolic compounds and benzo[a]pyrene, are efficiently adsorbed by PAF-6 MNPs, and over 50% of such toxins are removed. Theoretical computations were performed to understand the molecular interaction mechanism between PAF-6 and such analytes. The results demonstrate that the PAF-6 MNPs sorbents show excellent adsorption of phenols, polycyclic aromatic hydrocarbons and nitroaromatics based on multiple π-π stacking and hydrogen-bond interactions. These results suggest that the PAF-6 MNPs can be applied to extraction, removal and determination of diverse trace multi-target analytes from complex media.

Urinary metal and metalloid biomarker study of Henoch-Schonlein purpura nephritis using inductively coupled plasma orthogonal acceleration time-of-flight mass spectrometry.

To obtain a better understanding as to whether concentration alterations of metals and metalloids in urine were related to Henoch-Schonlein purpura nephritis (HSPN), the profiles of as many as 29 elements in urine were compared among three groups, the Henoch-Schonlein purpura (HSP), HSPN and a healthy control group. To this end, a reliable method has been developed for the simultaneous quantification of multiple elements including Li, Be, B, Al, Sc, Ti, V, Cr, Mn, Fe, Ni, Co, Cu, Zn, Ga, Ge, As, Se, Rb, Sr, Mo, Cd, Sn, Sb, Cs, Ba, Tl, Pb and Bi in urine using inductively coupled plasma orthogonal acceleration time-of-flight mass spectrometry (ICP-oa-TOF-MS). The process of sample pre-treatment used a direct 20-fold dilution method with centrifuged urine. The internal standard element used for quantification was 103Rh, and 1,4-butanediol was chosen as a matrix matching reagent. The method detection limits of these 29 elements were in the range of 0.04-12ngmL-1. Results of statistical analysis revealed that the concentrations of 15 elements and the element homeostasis were significantly different among these three groups. Our study provides a potential method for HSPN metal and metalloid biomarker discovery.

SPE-UHPLC-FLD Method for the Simultaneous Determination of Five Anthraquinones in Human Urine Using Mixed-Mode Bis(tetraoxacalix[2]arene[2]triazine) Modified Silica as Sorbent.

The five anthraquinones compounds (including aloe-emodin, emodin, physcion, chrysophanol, and rhein) are regarded as the main effective ingredients in rhubarb (Dahuang in Chinese, one of the commonly used Chinese herbal medicines). In this work, a simple and effective solid phase extraction (SPE) method based on bis(tetraoxacalix[2]arene[2]triazine) modified silica gel as adsorbent was developed. Coupled with UHPLC-FLD, the developed method was successfully applied for the measuring of main anthraquinones in human urine after oral administration of the extracts of rhubarb. To obtain the highest recoveries of the five anthraquinones in the SPE process, the main parameters which may affect extraction efficiency were optimized. The optimized sorbent amount, sample loading pH, sample loading rate, washing solution, and eluent condition were obtained. The developed method showed good linearity in 0.012-1.800 µg mL-1 for the five anthraquinones with correlation coefficients more than 0.9993. The investigated LOD values ranged from 3.9 to 5.7 ng mL-1, while the LOQs were between 12.0 and 18.2 ng mL-1. The recoveries of the method were also investigated, which were in the range of 94.8-106.6%. The application of the mixed-mode SPE materials in the proposed method was feasible and simple, and suitable for the enrichment of anthraquinones in urine samples.

Determination of γ-hydroxybutyrate in human urine samples by ion exclusion and ion exchange two-dimensional chromatography system.

A two-dimensional ion chromatography system was developed for the determination of γ-hydroxybutyrate (GHB) in human urine samples. Ion exclusion chromatography was used in the first dimensional separation for elimination of urine matrices and detection of GHB above 10mgL-1, ion exchange chromatography was used in the second dimensional separation via column-switching technique for detection of GHB above 0.08mgL-1. Under the optimized chromatographic conditions, the ion exclusion and ion exchange chromatography separation system exhibited satisfactory repeatability (RSD<3.1%, n=6) and good linearity in the range of 50-1000mgL-1 and 0.5-100mgL-1, respectively. By this method, concentrations of GHB in the selected human urine samples were detected in the range of 0-1.57mgL-1. The urine sample containing 0.89mgL-1 GHB was selected to evaluate the accuracy; the spiked recoveries of GHB were 95.9-102.8%. The results showed that the two-dimensional ion chromatography system was convenient and practical for the determination of GHB in human urine samples.