Simultaneous kinetic method for the determination of vitamin C, citrate and oxalate employing the Kalman filter

A kinetic method for the determination of vitamin C, citrate and oxalate in their mixture is described. The method involves the use of cerium(IV) as an oxidant and measurement of reaction rates spectrophotometrically by following the decrease in absorbance of cerium(IV) at 410 nm. The adaptive Kalman filter was used for data manipulation and analysis. It is shown that the use of the Kalman filter is superior to the classical differential kinetic methods owing to its suitability for the determination of analytes that react with a single reagent and exhibit a reaction rate constant ratio of less than 1.5. The results obtained were found to be highly precise and accurate even in the presence of some expected interferents.     

Simultaneous determination of guanine, uric acid, hypoxanthine and xanthine in human plasma by reversed-phase high-performance liquid chromatography with amperometric detection

A reversed-phase high-performance liquid chromatographic method with amperometric detection is described for the separation and quantification of uric acid, guanine, hypoxanthine and xanthine. The isocratic separation of a standard mixture of the compounds was achieved in 5 min on a Spherisorb 5 C18 reversed-phase column, with a mobile phase of NaH2PO4 (300 mmol dm-3, pH 3.0)-methanol-acetonitrile-tetrahydrofuran (97.8 + 0.5 + 1.5 + 0.2). Uric acid, guanine, hypoxanthine and xanthine were completely separated, with detection limits in the range 2-20 pmol per injection. The effect of pH and the composition of the mobile phase on the separation are described. The hydrodynamic voltammograms of these compounds were recorded at a glassy carbon electrode. The linear range of the calibration graph for each compound was: uric acid, 1-5000 mumol dm-3; guanine, 0.5-2000 mumol dm-3; hypoxanthine, 0.1-500 mumol dm-3 and xanthine, 0.5-5000 mumol dm-3. The within- and between-day precision was good. The uric acid and hypoxanthine content in human plasma was measured using the proposed method. Good recoveries of uric acid (97.9-103%), hypoxanthine (98.0-99.2%), guanine (96.0-98.3%) and xanthine (96.0-102%) were obtained from human plasma. The results of electrochemical detection were in good agreement with those of UV detection.     

离子色谱法同时测定环保型电镀抛光液中氨基磺酸和DL-苹果酸

采用Metrosep A Supp5150阴离子分离柱,以3.2mmol/L Na2CO3-1.0mmol/L NaHCO3为淋洗液,氨基磺酸和DL-苹果酸可得到很好的分离,使用离子色谱为检测手段可实现环保型电镀抛光液中氨基磺酸和DL-苹果酸的同时测定。两者的线性范围分别为1.6～175mg/L和3.2～175mg/L;检出限分别为0.4mg/L和0.8mg/L;相对标准偏差分别小于1.1%和0.9%(n=9);氨基磺酸的加标回收率在97%～103%之间,DL-苹果酸的加标回收率在93%～105%之间。方法用于新型电镀抛光液中氨基磺酸和DL-苹果酸的同时测定,具有很好的实用性。对实现抛光液中氨基磺酸和DL-苹果酸的在线监测具有实际意义。     

Simultaneous determination of p-aminohippuric acid, acetyl-p-aminohippuric acid and iothalamate in human plasma and urine by high-performance liquid chromatography

A sensitive and specific high-performance liquid chromatographic assay was developed for the simultaneous determination of p-aminohippuric acid (PAH), acetyl-p-aminohippuric acid (aPAH), and iothalamate in human plasma and urine. Plasma samples were prepared by protein precipitation with acetonitrile followed by evaporation, reconstitution in mobile phase, and injection onto a C18 reversed-phase column. Urine samples were diluted with 3 volumes of mobile phase prior to injection. Column effluent was monitored by UV detection at 254 nm. The lower limits of quantification in plasma were 0.5 mg/l for PAH and aPAH, and 1.0 mg/l for iothalamate. The within-day and between-day coefficients of variation in plasma and urine were < or =7.8% for all analytes. This method is well suited for renal function studies using iothalamate and PAH, whether administered as a bolus dose or by continuous infusion, to measure glomerular filtration rate and effective renal plasma flow, respectively.     

Simultaneous determination of phospholipid hydroperoxides and cholesteryl ester hydroperoxides in human plasma by high-performance liquid chromatography with chemiluminescence detection

A method was developed for the simultaneous determination of phosphatidylcholine hydroperoxides (PCOOH) and cholesteryl ester hydroperoxides (CEOOH). Lipid hydroperoxides (LOOH) were quantitatively extracted from human plasma with a mixture of n-hexane and ethyl acetate, and separated by column-switching high-performance liquid chromatography using one aminopropyl column and two octyl columns followed by chemiluminescence detection. LOOHs could be completely separated from each other and detected at picomole levels. The results of method validation tests were satisfactory. This method was then applied to determine LOOH in normal human plasma; the levels of PCOOH and CEOOH found were 36.0+/-4.0 nM (mean+/-S.D., n=6) and 12.3+/-3.1 nM (mean+/-S.D., n=6), respectively.     

离子对反相色谱法同时测定钢铁及合金中锆和铪

选用Zr(Hf)-F-5 Br PADAP体系进行锆和铪的高灵敏度显色反应,确定了柱前衍生、色谱分离及测定的最佳条件。考察了5-Br-PADAP的浓度、pH值、流动相、流动相的组成及显色时间对锆和铪络合物及色谱分离的影响。选用乙腈和水的混合液为流动相,流动相中对离子试剂NaF的浓度为1.5×10-4～3×10-4mol/L,pH4.2±0.2;用C18作为分离柱,峰面积与样品浓度线性范围为Zr:1.6～200μg/L,Hf:1.3～200μg/L,分离度1.84。方法的检测限为Zr:12.1μg/g,Hf     

Simultaneous determination of epirubicin, doxorubicin and their principal metabolites in human plasma by high-performance liquid chromatography and electrochemical detection

A method is described which permits the trace analysis of 10 chlorobenzenes in aqueous samples. Chlorobenzenes were extracted from water samples by solid-phase extraction with a C18 cartridge and analysis was carried out by gas chromatography-mass spectrometry in the selected-ion monitoring mode. The recovery and precision of the method were evaluated by extraction of spiked reagent-grade water at concentration levels of 0.1, 1.0 and 10.0 micrograms/l. This method was applied to the determination of chlorobenzenes in tap, ground and river water. By preparing 200 ml of environmental water samples, the detection limits of the compounds studied were in the range of 0.010-0.042 microgram/l.     

Simultaneous determination of coumarin, 7-hydroxycoumarin and 7-hydroxycoumarin glucuronide in human serum and plasma by high-performance liquid chromatography

Persistent viruses occur intracellularly in brown algae, specifically the Ectocarpales, and as reported here in the genus Feldmannia. Feldmannia species are small (1 mm-several cm), filamentous forms with single-celled meiotic sporangia that normally produce haploid zoospores. In the isolate reported here, spores were not observed in the sporangia but rather numerous (approximately 10(6) per cell) polyhedral viruses are formed in their place. Two dsDNA genome classes of 158 and 178 kbp, with two restriction site variants of each, are described. The individual abundance of each genome in viral preparations is affected by culture temperature. A cosmid library was used to generate circular restriction enzyme (BamHi, Noti, and Psti) site maps.     

硫酸钡微粒同步散射光谱法测定水泥样品中硫酸根离子

0.1mol/LHCl-4.0mg/mLTritonX 100-0.05mol/LBaCl2溶液体系的同步散射很弱,加入SO2-4浓度在4后,形成的(BaSO4)n微粒在470nm处产生一较强同步散射峰,SO2-2.0～80μg/mL浓度范围内与I470呈线性关系,据此建立了一个简便灵敏的测定水泥样品中硫酸根的同步散射光谱分析新方法。其相对标准偏差(RSD)在1.3%～2.7%之间,回收率在98.4%～101%之间。研究结果表明,由于静电引力SO2-4与Ba2+之间可形成BaSO4分子,而BaSO4分子间     

Simultaneous determination of N,N',N"-triethylenethiophosphoramide, cyclophosphamide and some of their metabolites in plasma using capillary gas chromatography

A sensitive assay for the simultaneous determination of N,N',N"-triethylenethiophosphoramide (thioTEPA), its metabolite N,N',N"-triethylenephosphoramide (TEPA), cyclophosphamide (CP) and its metabolite 2-dechloroethylcyclophosphamide (2-DCE-CP) in plasma has been developed and validated. The analytes were determined using gas chromatography with nitrogen/phosphorus selective detection after liquid-liquid extraction with chloroform using 100 microl of plasma. Diphenylamine (for TEPA, thioTEPA and 2-DCE-CP) and imipramine (for CP) were used as internal standards. The limits of quantitation for thioTEPA, TEPA, CP and 2-DCE-CP were 5, 5, 50 and 250 ng/ml, respectively. Linear calibration curves were observed over two decades of concentration. Accuracy, within-day and between-day precision were less than 13% for all analytes. Stability of the analytes proved to be satisfactory for at least 1 month, stored at -70 degrees C. Analysis of samples obtained from patients receiving cyclophosphamide, thioTEPA and carboplatin in a high-dose regimen demonstrated the applicability of the assay.     

Simultaneous determination of GTS-21 and its metabolite in rat plasma by high-performance liquid chromatography using solid-phase extraction

It has been suggested that GTS-21 can improve the learning deficits and inhibit the neuro-degeneration in patients with Alzheimer's disease. This paper describes a reversed-phase high-performance liquid chromatographic assay with visible detection at 405 nm for determination of GTS-21 and its metabolite, 4-hydroxy-GTS-21 in rat plasma. The method uses solid-phase extraction with a Bond Elut C18 column. A quantitation limit of 1.0 ng/ml was achieved using 0.5 ml of rat plasma. In the validation study, the coefficients of variation and the relative errors of each compound were less than 10%. Also freeze-thaw and storage stability were confirmed. This method has proved to be applicable to the pharmacokinetic study of GTS-21 in rats.     

Simultaneous determination of mycophenolic acid and its glucuronide in human plasma using a simple high-performance liquid chromatography procedure

We describe a reversed-phase HPLC method for determination of total mycophenolic acid (MPA), its free concentration (MPAf), and the glucuronide metabolite (MPAG), based on simple sample preparation and gradient elution chromatography. The compounds were quantified in parallel by absorbance at 254 nm and 215 nm in the internal standard mode. Linearity was verified up to 50 mg/L for MPA and up to 500 mg/L for MPAG (r >0.999). Detection limits at 215 and 254 nm were, respectively, 0.01 and 0.03 mg/L for MPA, and 0.03 and 0.1 mg/L for MPAG. The recovery of MPA was 95-106%; recovery of MPAG was 96-106%. The imprecision (CV) for MPA (0.2-25 mg/L) was <8.4% (254 nm) and <4.4% (215 nm) within day (n = 12) and <9.2% (254 nm) and <6.2% (215 nm) between days (n = 12). The imprecision for MPAG (10-250 mg/L) was <4.9% (254 nm) and <3.4% (215 nm) within day, and <6.1% (254 nm) and <5.9% (215 nm) between days. For quantification of MPAf, 100 microL of ultrafiltrate was applied directly to the column. The detection limit was 0.005 mg/L at 215 nm and 0.015 mg/L at 254 nm. In the range between 18-210 microg/L, the within-day CVs were <11.8% (n = 12) and the between-day CVs were <15.8% (n = 12).     

纳米二氧化钛中硫酸根的离子色谱法测定

建立了纳米二氧化钛（TiO2）中硫酸根(SO42-) 的离子色谱检测方法。通过研究纳米二氧化钛研磨、静置、加热、超声波等前处理条件,样品的溶解介质和取样量对样品测定值的影响优化了样品的检测条件。色谱条件为：Metrosep A SUPP5（250 cm）阴离子交换柱, Na2CO3／NaHCO3 淋洗液,流速为1 mL/min,MSM 抑制器和电导检测器。研究结果表明：采用10 g/L EDTA溶液和 pH10 缓冲溶液在常温静置浸提0.01 g 纳米TiO2,然后在Na2CO3／NaHCO3的淋洗液条件下进行抑制分离检测,获得了理想的检测效果。样品的加标回收率为93%~95%,测定结果的相对标准偏差小于3%（n=8）。     

Simultaneous determination of benzene, toluene, ethylbenzene, and xylenes in urine by thermal desorption-gas chromatography

Amphotericin B (AmB)-resistant Leishmania donovani promastigotes were selected by increasing drug pressure, and their biological features were compared with those of the wild-type parent strain. The 50% inhibitory concentration for resistant cells was 20 times higher than that for the wild-type. Resistance was stable after more than 40 passages in drug-free medium, and resistant promastigotes were infective to macrophages in vitro but lost their virulence in vivo. They had 2.5 times longer generation time, decreased AmB uptake, and increased AmB efflux in comparison to the wild type. Fluorescence measurement with a specific plasma membrane probe, 1-[4-(trimethylammonio)-1,6-diphenylhexa]-1,3,5-triene, showed increased membrane fluidity in drug-resistant promastigotes. Analysis of lipid composition showed that in resistant cells saturated fatty acids were prevalent, with stearic acid as the major fatty acid, and the major sterol was an ergosterol precursor, the cholesta-5, 7, 24-trien-3beta-ol and not ergosterol as in the AmB-sensitive strain.     

Simultaneous determination of spironolactone and its metabolites in human plasma

This study describes a specific, precise, sensitive and accurate method for determination of unchanged spironolactone and its major active metabolites in human plasma. After one-step liquid-liquid extraction, analysis of the parent drug and its metabolites was performed in one chromatographic run, using a high performance liquid chromatography (HPLC) method with a programmed switchover of the UV wavelength. Spironolactone and 7 alpha-thiomethyl-spironolactone were detected at 245 nm, while canrenone and internal standard were detected at 280 nm. The column used was an S5 ODS2 (500 mm x 4.6 mm i.d.). The mobile phase was a mixture of acetonitrile-aqueous orthophosphoric acid (pH 3.4). Chromatographic separations were performed at 5 degrees C. The standard curves were linear over the range 10-400 ng ml-1 for spironolactone and 10-600 ng ml-1 for 7 alpha-thiomethyl-spironolactone and canrenone. The precision and accuracy of the method were confirmed by relative standard deviations below 10% for different concentrations, except for the concentration equal to the quantitation limit, where these parameters ranged from 12-15%. The recovery was above 80% for all investigated compounds and for the internal standard. The assay proved to be suitable for pharmacokinetic studies of spironolactone.     

离子色谱法测定电子工业用高纯盐酸中痕量硫酸根离子

电子纯试剂(高纯盐酸)是电子化学品技术领域的关键性材料,建立一种电子工业用高纯盐酸中痕量硫酸根离子含量的离子色谱检测方法,对微电子技术发展具有重要意义.在高温条件下充分蒸发浓缩样品以除去其中的氯化氢和水,采用固相萃取柱银柱和钠柱分别除去除氯离子和金属离子,用超低压浓缩柱在线预富集,高容量阴离子分析柱分离测定,实现了离子...     

Simultaneous determination of catechins in human saliva by high-performance liquid chromatography

Green tea extracts have been suggested to possess a preventive effect against dental caries. A quantitative method for their anticariogenic substances, catechins, was developed to evaluate their concentrations in human saliva after mouthrinsing with green tea extract. Salivary catechins were extracted to the organic phase after forming a complex with diphenylborate and an ion-pair with tetra-n-butylammonium, and then back-extracted to the acidic aqueous phase. The extract was analyzed by high-performance liquid chromatography using diode array detection at absorption wavelengths ranging from 269 to 278 nm. In reversed-phase chromatography by a gradient elution, eight catechins originating from green tea and an internal standard were separated in 15 min without interfering peaks. All the catechins were simultaneously and selectively determined in the concentration range 0.05-25.0 microg/ml. In replicate spiking experiments with standards, the mean recovery ranged between 86 and 99%, and both intra- and inter-assay C.V.s were within 2.3%. When mouthrinsing with an aqueous solution of green tea extract (5.0 mg/ml) containing eight catechins, the quantitative results revealed that each catechin was retained at microg/ml levels in saliva for up to 60 min.     

Simultaneous determination of azathioprine and 6-mercaptopurine by high-performance liquid chromatography

We investigated whether linear whole-body acceleration along the interaural y-axis influenced the concurrent perception of visual motion direction as has been shown for angular accelerations. A sled running on air bearings along a 7.5-m track was used to accelerate 18 subjects at two different linear accelerations. These young, healthy volunteers, aged 25.50 +/- 7.38 years, used a joystick to indicate whether or not they perceived visual motion to the left within a random-dot kinematogram continuously presented on a monitor moving with them. The percentage of coherently leftward moving pixels presented for a 640-ms period during acceleration was adjusted according to a Modified Binary Search (MOBS) procedure. Six conditions were tested, two acceleration levels of 1 and 2 m/s2 to both left and right with, at the higher acceleration, two different times of visual motion presentation. Conditions were sequenced by means of a 6 x 6 Latin square balanced for order and carry over. A MANOVA did not show any statistically significant effects either for the independent variables acceleration, velocity, and direction of motion of the sled or for their interactions. The results obtained are in clear contrast to those obtained under rotatory stimulation. We conclude that the otolithic contribution to vestibular-visual motion processing is negligible.     

Simultaneous determination of diphenhydramine, its N-oxide metabolite and their deuterium-labeled analogues in ovine plasma and urine using liquid chromatography/electrospray tandem mass spectrometry

Our studies on drug disposition in chronically instrumented pregnant sheep involve simultaneous administration of the antihistamine diphenhydramine (DPHM), its deuterated analogue ([2H10]DPHM) and their metabolites to the mother or the fetus via various routes. Such studies require sensitive and selective mass spectrometric methods for quantitation of these labeled and unlabeled compounds in order to assess comparative maternal and fetal drug metabolism. The objective of this study was to develop and validate a liquid chromatographic/tandem mass spectrometric (LC/MS/MS) method for the simultaneous quantitation of DPHM, its N-oxide metabolite and their deuterium-labeled analogues in ovine plasma and urine. Samples spiked with the analytes and the internal standard, orphenadrine, were processed using liquid-liquid extraction. The extract was chromatographed on a propylamino LC column and MS/MS detection was performed in the positive ion electrospray mode using multiple reaction monitoring. The linear concentration ranges of the calibration curves for the N-oxides and the parent amines were 0.4-100.0 and 0.2-250.0 ng ml-1, respectively. In validation tests, the assay exhibited acceptable variability (< or = 15% at analyte concentrations below 2.0 ng ml-1 and < 10% at all other concentrations) and bias (< 15% at all concentrations), and the analytes were stable under a variety of sample handling conditions. Using this method, the labeled and unlabeled N-oxide metabolite was identified in fetal plasma after DPHM and [2H10]DPHM administration. This method will be used further to examine the comparative metabolism of diphenhydramine to its N-oxide metabolite in the mother and the fetus.