sCD163, sCD28, sCD80, and sCTLA-4 as soluble marker candidates for detecting immunosenescence.

BACKGROUND: Inflammaging, the characteristics of immunosenescence, characterized by continuous chronic inflammation that could not be resolved. It is not only affect older people but can also occur in young individuals, especially those suffering from chronic inflammatory conditions such as autoimmune disease, malignancy, or chronic infection. This condition led to altered immune function and as consequent immune function is reduced. Detection of immunosenescence has been done by examining the immune risk profile (IRP), which uses flow cytometry. These tests are not always available in health facilities, especially in developing countries and require fresh whole blood samples. Therefore, it is necessary to find biomarkers that can be tested using stored serum to make it easier to refer to the examination. Here we proposed an insight for soluble biomarkers which represented immune cells activities and exhaustion, namely sCD163, sCD28, sCD80, and sCTLA-4. Those markers were reported to be elevated in chronic diseases that caused early aging and easily detected from serum samples using ELISA method, unlike IRP. Therefore, we conclude these soluble markers are beneficial to predict pathological condition of immunosenescence. AIM: To identify soluble biomarkers that could replace IRP for detecting immunosenescence. CONCLUSION: Soluble costimulatory molecule suchsCD163, sCD28, sCD80, and sCTLA-4 are potential biomarkers for detecting immunosenescence.

Dysfunctional energy and future perspective of low dose H2O2 as protective agent in neurodegenerative disease.

The number of people with neurodegenerative disease continues to increase every year. A new perspective is needed to overcome this disease. In this review, researchers collected information about dysfunctional energy in neurodegenerative diseases driven by mitochondria. Mitochondrial dysregulation can cause damage to the neuron system. The increase in the amount and interaction of α-synuclein with SAMM50 and GABARAPL1 in the mitochondria is one of the factors causing neurodegenerative disease. As an energy provider in the body, the existence of harmonization in the regulation of mitochondria, specifically the mitochondrial outer membrane, is important. Low-dose hydrogen peroxide (H2O2) has neuroprotective abilities to overcome the impairment function of mitochondria in neurodegenerative patients. Based on computational simulation of this case, it can be used as a basic concept for the development of the role of H2O2 in neurodegenerative diseases.

Green Synthesis of Silver Nanoparticles Coated by Water Soluble Chitosan and Its Potency as Non-Alcoholic Hand Sanitizer Formulation.

The synthesis of silver nanoparticles using plant extracts, widely known as a green synthesis method, has been extensively studied. Nanoparticles produced through this method have applications as antibacterial agents. Bacterial and viral infection can be prevented by use of antibacterial agents such as soap, disinfectants, and hand sanitizer. Silver nanoparticles represent promising hand sanitizer ingredients due to their antibacterial activity and can enable reduced use of alcohol and triclosan. This study employed silver nanoparticles synthesized using Kepok banana peel extract (Musa paradisiaca L.). Nanoparticle effectiveness as a hand sanitizer can be enhanced by coating with a biocompatible polymer such as chitosan. The characterization of silver nanoparticles was conducted using UV-Vis, with an obtained peak at 434.5 nm. SEM-EDX analysis indicated nanoparticles with a spherical morphology. Silver nanoparticles coated with chitosan were characterized through FTIR to verify the attached functional groups. Gel hand sanitizers were produced using silver nanoparticles coated with different chitosan concentrations. Several tests were undertaken to determine the gel characteristics, including pH, syneresis, and antibacterial activity. Syneresis leads to unstable gels, but was found to be inhibited by adding chitosan at a concentration of 2%. Antibacterial activity was found to increase with increase in chitosan concentration.

Quantification and Improvement of the Dynamics of Human Serum Albumin and Glycated Human Serum Albumin with Astaxanthin/Astaxanthin-Metal Ion Complexes: Physico-Chemical and Computational Approaches.

Glycated human serum albumin (gHSA) undergoes conformational changes and unfolding events caused by free radicals. The glycation process results in a reduced ability of albumin to act as an endogenous scavenger and transporter protein in diabetes mellitus type 2 (T2DM) patients. Astaxanthin (ASX) in native form and complexed with metal ions (Cu2+ and Zn2+) has been shown to prevent gHSA from experiencing unfolding events. Furthermore, it improves protein stability of gHSA and human serum albumin (HSA) as it is shown through molecular dynamics studies. In this study, the ASX/ASX-metal ion complexes were reacted with both HSA/gHSA and analyzed with electronic paramagnetic resonance (EPR) spectroscopy, rheology and zeta sizer (particle size and zeta potential) analysis, circular dichroism (CD) spectroscopy and UV-Vis spectrophotometer measurements, as well as molecular electrostatic potential (MEP) and molecular docking calculations. The addition of metal ions to ASX improves its ability to act as an antioxidant and both ASX or ASX-metal ion complexes maintain HSA and gHSA stability while performing their functions.

Cobalt (II)-Mediated Molecularly Imprinted Polymer as a Monolithic Stationary Phase for Separation of Racemic Citronellal by Liquid Chromatography.

A metal-mediated molecularly imprinted polymer (MMIP) monolithic column was prepared as the stationary phase for high-performance liquid chromatography (HPLC) and applied to the enantiomeric separation of rac-citronellal. MMIP column was prepared through in situ copolymerizations with the ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate/[BMIM][BF4] as the primary pore-forming agent and cobalt(II) acetate as the metal pivot. Interactions between polymer components in the synthesized monolith were assessed by FTIR to identify the functional groups. The monolith morphology was characterized with SEM, and the template removal was detected by UV Spectrophotometry at 253 nm. In this study, (R)-(+)-citronellal was used as a template, whereas 4-vinylpyridine (4-VP) was employed as the functional monomer with two monomer crosslinkers, trimethylolpropane trimethacrylate (TRIM), and ethylene glycol dimethacrylate (EDMA). The ternary mixture of porogenic solvent consisted of [BMIM][BF4], dimethylformamide (DMF), and dimethyl sulfoxide (DMSO) with the applied ratio of 1 : 1:1 (v/v) and 10 : 1:5 (v/v) for the preparation of MMIP using TRIM and EDMA crosslinkers, respectively. Co(II) ion was added to the porogenic solvent before being mixed with the functional monomer and the crosslinker mixtures. Separating the rac-citronellal was achieved on the synthesized MMIP, showing better selectivity than the monolithic metal-mediated nonimprinted polymer (MNIP), nonimprinted polymer (NIP), and molecularly imprinted polymer (MIP).

Recent advances in nephropathy biomarker detections using paper-based analytical devices.

Nephropathy or kidney disease involves the deterioration of kidney functions, causing severe diseases, such as proteinuria, chronic kidney diseases, and kidney failure. Currently, nephropathy that develops into kidney failure is increasing globally, as indicated by the increasing number of patients undergoing hemodialysis. Some developed analytical methods for nephropathy using albumin, creatinine, uric acid, and the urinary albumin-to-creatine ratio biomarkers, including spectrophotometry, turbidimetric immunoassay, and ELISA, have been reported so far, providing good accuracy and precision. However, WHO has established guidelines for developing diagnostic tools that meet several criteria: Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, Delivered to those who need it. This means that nephropathy detection can be carried out using a simple method compatible with point-of-care that allows independent urine analysis by patients. For this purpose, the use of paper-based analytical devices (PADs) as an alternative platform for the detection of albumin, creatinine, uric acid, and the urinary albumin-to-creatine ratio were reviewed.

An application study of membraneless-gas separation microfluidic paper-based analytical device for monitoring total ammonia in fish pond water using natural reagent.

The membraneless-gas separation microfluidic paper-based analytical device (ML-GS µPAD), consisting of donor, spacer, and acceptor layers, was developed to monitor total ammonia in fish pond water. The principle of the analysis involved the addition of sodium hydroxide solution to the sample zone in the donor layer containing ammonia/ammonium, and the produced ammonia gas diffuses through the spacer to the detection zone in the acceptor layer containing red rose extract to produce a color change from pink to blue corresponding to the ammonia/ammonium concentration. Under optimum conditions, the proposed method provided good linearity of ammonia in the range concentration of 0-100 mg L-1 (R2 = 0.9993) with LOD and LOQ of 2.25 and 7.51 mg L-1, respectively. This method was successfully applied to fish pond water samples without significant influence of interfering compounds with recoveries in the range of 103-110%, indicating good selectivity and accuracy of the proposed method.

An experimental design approach for the optimization of scopoletin extraction from Morinda citrifolia L. using accelerated solvent extraction.

Scopoletin is regarded as a major constituent of noni (Morinda citrifolia L), which contributes to the antioxidative, anti-inflammatory, immunomodulatory, and hepatoprotective properties in noni. It is also suggested as a marker for identification and quality control of noni and its derivative products. With the increasing interest in noni due to its health benefits and therapeutic effects, it is important to establish a reliable extraction and analysis method to determine scopoletin content in noni samples. The present study proposes the use of accelerated solvent extraction (ASE) to extract scopoletin from noni, followed by detection using HPLC-DAD for rapid identification and quantification of scopoletin. The optimum operating conditions of ASE were investigated using response surface methodology (RSM), using a three factors central composite design. It was found that the optimum scopoletin yield was achieved by performing the extraction at an elevated temperature of 60 °C for 12 min, using ethanol as extraction solvent with solid to solvent ratio of 1:30 (w/v). The analytical method validation was carried out under optimum conditions. The results indicate that the proposed ASE-HPLC-DAD method was adequately sensitive for the quantification of scopoletin in extracts with limit of detection of 0.17 µg/g. The presented method also exhibits excellent linearity from 0.54 to 120.10 µg/g with R2 0.9995, high precision with RSD lower than 2% for intra-day and inter-day replication, and good recovery (99.88%). The established method was also successfully applied for scopoletin determination in noni product samples. The developed method provides a rapid and reliable method for the identification and quantification of scopoletin in noni samples that is suitable for routine procedures.

Preparation of Metal-Immobilized Methacrylate-Based Monolithic Columns for Flow-Through Cross-Coupling Reactions.

With the aim of developing efficient flow-through microreactors for high-throughput organic synthesis, in this work, microreactors were fabricated by chemically immobilizing palladium-, nickel-, iron-, and copper-based catalysts onto ligand-modified poly(glycidyl methacrylate-co-ethylene dimethacrylate) [poly(GMA-co-EDMA)] monoliths, which were prepared inside a silicosteel tubing (10 cm long with an inner diameter of 1.0 mm) and modified with several ligands including 5-amino-1,10-phenanthroline (APHEN), iminodiacetic acid (IDA), and iminodimethyl phosphonic acid (IDP). The performance of the resulting microreactors in Suzuki-Miyaura cross-coupling reactions was evaluated, finding that the poly(GMA-co-EDMA) monolith chemically modified with 5-amino-1,10-phenanthroline as a binding site for the palladium catalyst provided an excellent flow-through performance, enabling highly efficient and rapid reactions with high product yields. Moreover, this monolithic microreactor maintained its good activity and efficiency during prolonged use.

Immobilization of trypsin onto porous methacrylate-based monolith for flow-through protein digestion and its potential application to chiral separation using liquid chromatography.

Monolithic columns for analytical applications have attracted the researcher's attention. In this work, the laboratory-made organic-polymer monolithic column is modified with trypsin and further applied as a nanobiocatalyst microreactor and a stationary phase for separating chiral compounds by liquid chromatography. The monolith was synthesized by in-situ copolymerization of glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EDMA) or trimethylolpropane trimethacrylate (TRIM) as a crosslinking agent, with porogen of 1,4-butanediol/propanol/water (4:7:1 v/v) and AIBN as the radical polymerization initiator inside PEEK and silicosteel tubings (1.0 mm i.d × 100 mm) at 60 °C for 12 h. A total monomer ratio (%T) and crosslinking agent (%C) of 40:25 and 28:12 were applied to prepare poly-(GMA-co-EDMA) and poly-(GMA-co-TRIM), respectively. The produced monoliths were further modified by introducing trypsin (10 mg/L) through the ring-opening reaction of the epoxide group existing in the monolithic column. The trypsin-immobilized poly-(GMA-co-EDMA) monolithic column was applied as the nanobiocatalyst microreactor for online/flow-through and rapid digestion of ß-casein sample into its peptide fragments. The trypsin-immobilized poly-(GMA-co-TRIM) column has potential application to be used as the HPLC stationary phase for the separation of R/S-citronellal enantiomers.

DFT and molecular dynamics studies of astaxanthin-metal ions (Cu2+ and Zn2+) complex to prevent glycated human serum albumin from possible unfolding.

Glycated human serum albumin (gHSA) undergoes conformational changes of proteins caused by free radicals. The glycation process results in a reduced ability of albumin as an endogenous scavenger in diabetes mellitus type 2 (T2DM) patients. Astaxanthin (ASX) has been shown to prevent gHSA from experiencing unfolding events and improve protein stability of gHSA and HSA through molecular dynamics. In this study, astaxanthin is complexed with transition metal ions such as copper (Cu2+) and zinc (Zn2+) in two modes (M) and (2M). Complexing astaxanthin with Cu2+ and Zn2+ is expected to increase astaxanthin's ability as an endogenous scavenger than in native form. This research aims to characterize the antiradical property of ASX, ASX-Cu2+ and ASX-2Cu2+, ASX-Zn2+, and ASX-2Zn2+ with density functional theory (DFT) and to compare the capability to prevent conformational changes on glycated albumin through molecular dynamics simulation. DFT as implemented in Gaussian 09W, was used for all calculations. Analysis of data using GaussView 6.0. LANL2D2Z basis set and B3LYP density functional used for frequency analysis and optimization. The AutoDock Vina implemented in PyRx 0.8 is used to and receptor-ligand interactions analysis with the DS 2016 Client. YASARA for molecular dynamic simulation with 15,000 ps as running time. DFT analyzes such as energy gaps, HOMO, and LUMO patterns and electronic properties have shown that ASX-metal ions complex is better than ASX in native state as antioxidants. These results are also supported by the molecular dynamics simulation (RMSD backbone, RMSDr, RMSFr, and movie visualization), where the addition of ASX-metal ions complex on gHSA are better than ASX as a single compound in preventing gHSA from possible unfolding and maintaining protein molecule stability.

Recent Advances of Hepatitis B Detection towards Paper-Based Analytical Devices.

Hepatitis B virus (HBV) still remains a major global public health problem. One-half to one-third of the total HBV infected people died due to late detection of HBV. Serological antigen and viral HBV detections can help in the diagnosis, referral, and treatment of HBV. Available methods for HBV detection mostly used bulky instruments. Miniaturization of devices for HBV detection has been started by narrowing down the size of the devices. Several methods have also been proposed to increase the selectivity and sensitivity of the miniaturized methods, such as sandwich recognition of the biomarkers and the use of nano- to micro-sized materials. This review presents recent HBV detections in the last two decades from laboratory-based instruments towards microfluidic paper-based analytical devices (µPADs) for point-of-care testing (POCT) purposes. Early and routine analysis to detect HBV as early as possible could be achieved by POCT, especially for areas with limited access to a central laboratory and/or medical facilities.

Preparation and Kinetic Studies of Cross-Linked Chitosan Beads Using Dual Crosslinkers of Tripolyphosphate and Epichlorohydrin for Adsorption of Methyl Orange.

Preparation of cross-linked chitosan beads using dual crosslinkers of tripolyphosphate (TPP) and epichlorohydrin (ECH) for the adsorption and kinetic studies of methyl orange (MO) had been carried out. FTIR spectra showed that TPP could act as the protecting agent of the NH2 group of chitosan and ECH reacted with the primary hydroxyl group of chitosan. Various concentrations of TPP, ECH, and immersing time in the TPP solution for bead formation were studied. The effect of pH and kinetics of adsorption were investigated to define the mechanism of adsorption and rate-limiting step. As a result, pH 3, 10% (w/v) TPP, 5% (v/v) ECH, and 12 h immersing time in TPP were selected as the optimum conditions for preparing the beads as indicated by the highest adsorption amount of MO. The cross-linked chitosan beads' adsorption capacity for MO under optimum condition was found to be 79.55 mg/g with the adsorption rate constant (k) of 1.29 × 10-3/min. Furthermore, it was found that a low concentration of ECH could maintain the stability of chitosan in acidic conditions, whereas the concentration of TPP and immersing time controlled pore size and morphology of chitosan beads. The mechanism of adsorption of MO was controlled by the pore and rigidity of cross-linked chitosan beads. Bulk diffusion acted as a rate-limiting step, and a high concentration of MO inhibited diffusion and adsorption itself.

Green synthesis and characterization of copper nanoparticles using Piper retrofractum Vahl extract as bioreductor and capping agent.

The copper nanoparticles (CuNPs) have attracted much attention due to their application in diverse fields. The applications of CuNPs depend on their physical and chemical properties. This study presents the first report for the use of medicinal fruit extract of Piper retrofractum Vahl as an eco-friendly reagent in the synthesis of CuNPs using copper sulfate as a starting material. Piper retrofractum Vahl extract was employed as a bioreductor as well as a capping agent in the formation of CuNPs. The reaction process was assisted by sonication and stirring. The influences of extract concentration, pH, temperature, and reaction time on the size of CuNPs were studied in detail. The morphology and structure of synthesized CuNPs were characterized by UV-Vis, FT-IR, SEM-EDS, TEM, and XRD. The UV-Vis measurement showed the surface plasmon resonance (SPR) peak at 234-255 nm, whereas FTIR characteristic peaks of metal-oxygen (Cu-O) were confirmed in the range 550-570 cm-1 and Cu-O-H bonds led to bending absorptions in the region 870-880 cm-1. The synthesized CuNPs possess the spherical shapes and high content of copper (70.3%) as confirmed by SEM-EDS. From the TEM micrograph, it can be seen that the particle size distribution of CuNPs has a high uniformity with a size of 2-10 nm under the optimum condition. The crystalline nature of CuNPs as confirmed by XRD showed the crystallinity phase of 26.4%. The synthesized CuNPs have relatively good stability and could inhibit Escherichia coli and Staphylococcus aureus. The results proved that Piper retrofractum Vahl fruit extract could be applied for a greener synthesis of CuNPs with high uniformity of particle sizes.

Synthesis and Characterisation of a Monolithic Imprinted Column Using a Methacrylic Acid Monomer with Porogen Propanol for Atenolol Analysis.

A monolithic imprinted atenolol column was constructed by in situ polymerisation using a methacrylic acid monomer and a 1 : 1 (v/v) porogen of propanol: toluene with two template: monomer: crosslinker combinations, namely, MIP 1 (1 : 4 : 20) and MIP 2 (1 : 5 : 20). Physical characterisation of the monolithic columns consisted of permeability testing, Fourier transform infrared (FTIR) testing, surface area analysis (SAA), and scanning electron microscopy (SEM). The permeability value of four monolithic columns was in the good category: MIP 1 (24.01 mD), NIP 1 (56.43 mD), MIP 2 (23.03 mD), and NIP 2 (14.47 mD). The polymerisation process of these four monolithic imprinted columns was carried out perfectly, as shown by the absence of vinyl groups (1000 cm-1 and 900 cm-1) during FTIR testing. Based on SAA testing, the pores of the four polymers were classified as mesopores. The best monolithic column was MIP 1, as seen from the intercolumn and intracolumn reproducibility values and a % RSD <2.0%. The MIP 1 column was selective towards atenolol, as seen from the selectivity factor, imprinting factor (IF), and resolution (Rs) values. The IF values of MIP 1 were atenolol (204.62), metoprolol (3.36), and propranolol (1.27). The Rs value between atenolol and the analogue compounds was 7.23. The MIP 1 column can be used for the analysis of atenolol in blood serum samples with an average percentage recovery rate of 94.88 ± 4.43%.

Preparation of methacrylate-based anion-exchange monolithic microbore column for chromatographic separation of DNA fragments and oligonucleotides.

In this paper, we report on the preparation of a microbore-scale (1 mm i.d.) anion-exchange monolithic column suitable not only for analytical purposes but also for potentially preparative applications. In order to meet the conflicting requirements of high permeability and good mechanical strength, the following two-step procedure was applied. First, an epoxy-containing monolith was synthesized by in situ copolymerization of glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) within the confines of a silicosteel tubing of 1.02 mm i.d. and 1/16" o.d. in the presence of a ternary porogenic mixture of 1-propanol, 1,4-butanediol, and water. The monolithic matrix was subsequently converted into weak anion-exchanger via the ring-opening reaction of epoxy group with diethyl amine. The dynamic binding capacity was 21.4 mg mL(-1) for bovine serum albumin (BSA) at 10% breakthrough. The morphology and porous structure of this monolith were assessed by scanning electron microscope (SEM) and inverse size exclusion chromatography (ISEC). To optimize the separation efficiency, the effects of various chromatographic parameters upon the separation of DNA fragments were investigated. The resulting monolithic anion exchanger demonstrated good potential for the separation of both single- and double-stranded DNA molecules using a gradient elution with NaCl in Tris-HCl buffer (20 mM). Oligodeoxythymidylic acids (dT(12)-dT(18)) were successfully resolved at pH 8, while the fragments of 20 bp DNA ladder, 100 bp DNA ladder, and pBR322-HaeIII digest were efficiently separated at pH 9.

Atomic mineral characteristics of Indonesian osteoporosis by high-resolution inductively coupled plasma mass spectrometry.

Clinical research indicates that negative calcium balance is associated with low bone mass, rapid bone loss, and high fracture rates. However, some studies revealed that not only calcium is involved in bone strengthening as risk factor of fracture osteoporosis. Thus, in this report, the difference of metallic and nonmetallic elements in osteoporosis and normal bones was studied by high-resolution inductively coupled plasma mass spectrometry (HR-ICP-MS). The influence of these elements on bone metabolic processes is also discussed. Inclusion criteria of bone samples consist of postmenopausal woman, trabecular bone fracture, normal and osteoporosis BMD value, and no history of previous disease. The results showed that the concentration of B, Al, S, V, Co, Mo, Te, Ba, La, Ni, As, and Ca/P ratio is higher in osteoporosis than normal. These atomic minerals have negative role to imbalance between bone resorption and bone formation activity. Conversely, concentrations of Na, Mg, P, K, Ca, Cr, Pd, Ag, Mn, Fe, Cu, Zn, Rb, Sr, Pb, and Se are lower in osteoporosis than in normal bones. Among these atoms, known to have important roles in bone structure, we found involvement of atomic mineral and calcium which are considerable to contribute to osteoporotic phenomena.

Chemical anchoring of lauryl methacrylate-based reversed phase monolith to 1/16″ o.d. polyetheretherketone tubing.

In this paper, we describe a method for the preparation of easy-to-use reversed-phase monolithic microbore columns. Polyetheretherketone (PEEK) tubing with an outer diameter of 1/16″ and an inner diameter of 1.0 mm was used as a column housing (empty column), and in it lauryl methacrylate (LMA) was copolymerized with ethylene dimethacrylate (EDMA). In order to chemically anchor the polymer monolith to the tube wall, the inner wall surface was pretreated by the following two-step procedure. (1) 50% sulfuric acid was filled into the PEEK tubing and left to stand for 6 h to generate sulfonate groups on the surface. (2) After washing with Milli-Q water, the sulfonated PEEK surface was brought into contact with 1 M glycidyl methacrylate in dichloromethane (or acetone) at 40°C for 4 h to introduce methacryloyl groups via the reaction of sulfonate groups and epoxy groups. Mechanical strength and column efficiency of the resulting monoliths were evaluated through the separation of a series of alkylbenzenes in acetonitrile-water (50:50, v/v) eluent over the flow rate range of 50-750 µL/min (corresponding to 1.7-25.5 mm/s). The poly(LMA-co-EDMA) monolith provided acceptable column efficiency of 2000 theoretical plates/10 cm (HETP value of 50 µm) for amylbenzene (separation factor k=40) and low flow resistance of 0.5 MPa/10 cm at a normal flow rate of 50 µL/min. The methacryloylated PEEK tubing tightly held the monolith, and the monolithic column exhibited good pressure resistance up to 15 MPa, allowing rapid separation at a 15-20 fold higher flow rate than normal.

Development of salt-tolerance interface for an high performance liquid chromatography/inductively coupled plasma mass spectrometry system and its application to accurate quantification of DNA samples.

Accurate quantification of DNA is highly important in various fields. Determination of phosphorus by ICP-MS is one of the most effective methods for accurate quantification of DNA due to the fixed stoichiometry of phosphate to this molecule. In this paper, a smart and reliable method for accurate quantification of DNA fragments and oligodeoxythymidilic acids by hyphenated HPLC/ICP-MS equipped with a highly efficient interface device is presented. The interface was constructed of a home-made capillary-attached micronebulizer and temperature-controllable cyclonic spray chamber (IsoMist). As a separation column for DNA samples, home-made methacrylate-based weak anion-exchange monolith was employed. Some parameters, which include composition of mobile phase, gradient program, inner and outer diameters of capillary, temperature of spray chamber etc., were optimized to find the best performance for separation and accurate quantification of DNA samples. The proposed system could achieve many advantages, such as total consumption for small amount sample analysis, salt-tolerance for hyphenated analysis, high accuracy and precision for quantitative analysis. Using this proposed system, the samples of 20 bp DNA ladder (20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 300, 400, 500 base pairs) and oligodeoxythymidilic acids (dT(12-18)) were rapidly separated and accurately quantified.

Multielement analysis of micro-volume biological samples by ICP-MS with highly efficient sample introduction system.

A method for multielement analysis of micro-volume biological sample by inductively coupled plasma mass spectrometry (ICP-MS) with a highly efficient sample introduction system was presented. The sample introduction system was the combination of (1) an inert loop injection unit and (2) a high performance concentric nebulizer (HPCN) coupled with a temperature controllable cyclone chamber. The loop injection unit could introduce 20 µL samples into the carrier liquid flow of 10 µL min(-1) producing a stable signal for 100s without any dilution. The injection loop is continuously washed with 0.1M HNO(3) carrier solution during the measurement, thereby much improving sample throughput. The HPCN is a triple tube concentric nebulizer, which can generate fine aerosols and provide a stable and highly measurement sensitivity in ICP-MS at a liquid flow rate less than 10 µL min(-1). With the combination of the chamber heating at 60°C, the sensitivity obtained with the proposed sample introduction system at the liquid flow rate of 10 µL min(-1) was almost the same as that with a common concentric nebulizer and cyclone chamber system at the liquid flow rate of 1 mL min(-1), though the sample consumption rate of the HPCN was two orders of the magnitude lower than that of the common nebulizer. The validation of the proposed system was performed by analyzing the NIST SRM 1577b Bovine Liver. The observed values for 12 elements such as Na, P, S, K, Ca, Mn, Fe, Co, Cu, Zn, Mo, Cd were in good agreement with their certified values and information value. Satisfactory analytical results for 14 elements such as Na, Mg, P, S, K, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Y, Ba in Escherichia coli sample were also obtained. The proposed sample introduction system was quite effective in the cases when only micro-volume of biological sample is available.