A thin-layer chromatographic method for the determination of acebutolol and its major metabolite in serum

A sensitive and specific thin-layer chromatographic method for the simultaneous determination of acebutolol [DL-1-(2-acetyl-4-_n-butyramidophenoxy)-2-hydroxy-3-isopropylaminopropane] and its major metabolite [DL-1-(2-acetyl-4-acetamidophenoxy)-2-hydroxy-3-isopropylaminopropane] is described. A 2-ml volume of serum with 350 ng of quinidine as internal standard was extracted at pH 10, the solvent was evaporated off and the residue was dissolved in 50 µ1 of methanol. A 10-µ1 volume of the solution was spotted on a thin-layer plate and after elution (ethyl acetate-methanol-ammonia, 75:20:5) the plate was dried at 90° for 15 min and, after cooling, dipped in a 10% paraffin wax solution. The fluorescence was measured using a spectrofluorimeter with a thin-layer scanning attachment. The peak-height ratios of acebutolol to internal standard and metabolite to internal standard were used to quantitate acebutolol and the metabolite, respectively.     

Assay of naproxen in rat serum by high-performance thin-layer chromatography/densitometry

A simple, sensitive, and reproducible high-performance thin-layer chromatographic method using densitometry is presented for the determination of naproxen in rat serum. Only 0.1 mL serum was used for extraction. Separations were performed on 10 x 10 cm plates coated with silica gel, with toluene-ethyl acetate-acetic acid (82 + 15 + 3) as the mobile phase. Benzophenone was used as the internal standard. Quantification was performed by densitometry at 260 nm. The response response for naproxen was linear (r = 0.992) over the range 2-100 mg/L. Method validation demonstrated good recoveries (92-96%), sensitivity (limit of quantitation, 1 mg/L), repeatability of sample application (4%), repeatability of the method (8%), and intermediate precision (5%). The procedure was applied to the quantitation of naproxen in rat serum.     

Quantitation of digitoxin and the bis- and monodigitoxosides of digitoxigenin in serum

A specific assay is described for measuring the concentration of digitoxin and the bis- and monoglycosides of digitoxigenin in serum. The procedure includes: (1) addition of a tracer amount of tritium labeled parent compound to the serum in order to measure percentage recovery; (2) solvent extraction to separate polar and non-polar metabolites; (3) reversed-phase thin-layer chromatography of the non-polar fraction to separate digoxigenins from digitoxigenins; (4) thin-layer chromatography to isolate digitoxin, and the bis- and monoglycosides of digitoxigenin; and (5) use of an 125I-radioimmunoassay to determine the concentration of the glycosides. Each of these three glycosides was administered intravenously to a normal subject, and the concentration of parent compound was measured in the serum at various times.     

Simultaneous determination of bergenin and gallic acid in Bergenia ligulata wall by high-performance thin-layer chromatography

A simple and reproducible high-performance thin-layer chromatographic method was developed for the simultaneous determination of bergenin and gallic acid in Bergenia ligulata. Water and methanol were used as the extracting solvents. The concentrations of bergenin and gallic acid in both of these solvents were found to be almost the same. The method involves separation of the components by thin-layer chromatography on a precoated Silica Gel 60 F254 plate with a solvent system of ethyl acetate-formic acid-acetic acid-water (100 + 11 + 11 + 27). The sensitivity of the method for bergenin was 0.30 microg, whereas for gallic acid it was 0.25 microg. The proposed method is precise and sensitive and can be used for the detection, monitoring, and simultaneous quantification of bergenin and gallic acid in B. ligulata.     

Quantitative in situ thin-layer chromatography of quinidine and salicylic acid in capillary blood.

A micro-method based on in situ thin-layer chromatography has been developed for the determination of quinidine and salicylic acid in 10-mu 1 samples of serum or plasma. The sample is applied directly to the silica gel layer without extraction, the proteins being precipitated on the chromatogram origin by means of ethanol. The chromatogram can be evaluated quantitatively by fluorescence scanning. Different pre-coated plates were investigated.     

基于一次性薄层色谱电极高选择电化学发光分析法测定Ni(II)的研究

基于一种新的电极制作方法, 研制了具有薄层色谱分离功能的一次性薄层色谱电极. 结合该电极的分离、保留分析物于电极表面一定空间区域的色谱分离能力与电极表面电化学发光信号的空间分辨能力, 实现了分析物的高效分离与原位高灵敏度电化学发光(ECL)检测, 建立了电化学发光分析方法与薄层色谱分离方法联用的新技术. 并以Ni^2＋离子为代表探讨了这一方法的可行性和分析特性. 在最佳的实验条件下, 该方法测定Ni^2＋离子的线性范围为5.0× 10^－9～5.0×10^－6 g/mL, 检出限为1.5×10^－9 g/mL, 相对标准偏差为2.8% (c＝1.0×10^－6 g/mL, n＝11).     

电极体插入式长光程可见-紫外薄层光谱电化学池设计——二茂铁在二氯乙烷及苯甲烷中的行为

长光程光谱电化学池灵敏度高,还可以研究电极表面的吸附与修饰,用处很广,但文献报导的各种设计,多为组合式,构造复杂,电极电位控制不够完善,在非水溶剂中,尤其在低介电常数溶剂中,得不到理想的结果。作者曾报导一电流平行流向短光程薄层光谱电化学池,在薄层池内外各设电位参比点,得到了近于理想的伏安特性、本文报告一整体式内参比点设置的长光程薄层光谱电化学池,使用插入式聚四氟乙烯薄层电极,以池壁自然面当光透窗,     

Effect of cellulase adsorption on the surface and interfacial properties of cellulose

The surface properties of several purified cellulose (Sigmacell 101, Sigmacell 20, Avicel pH 101, and Whatman CF 11) were characterised, before and after cellulase adsorption. The following techniques were used: thin-layer wicking (except for the cellulose Whatman), thermogravimetry, and differential scanning calorimetry (for all of the above celluloses). The results obtained from the calorimetric assays were consistent with those obtained from thin-layer wicking – Sigmacell 101, a more amorphous cellulose, was the least hydrophobic of the analysed celluloses, and had the highest specific heat of dehydration. The other celluloses showed less affinity for water molecules, as assessed by the two independent techniques. The adsorption of protein did not affect the amount of water adsorbed by Sigmacell 101. However, this water was more strongly adsorbed, since it had a higher specific heat of dehydration. The more crystalline celluloses adsorbed a greater amount of water, which was also more strongly bound after the treatment with cellulases. This effect was more significant for Whatman CF-11. Also, the more crystalline celluloses became slightly hydrophilic, following protein adsorption, as assessed by thin-layer wicking. However, this technique is not reliable when used with cellulase treated celluloses.     

Determination of the 7-amino-metabolites of the 7-nitro-benzodiazepines in human plasma by thin-layer chromatography

A sensitive thin-layer chromatographic method is described for the determination of the 7–amino-metabolites of flunitrazepam and clonazepam. The same procedure is applicable to other 7-nitro-benzodiazepines such as nitrazepam. A purified plasma extract of the 7-amino-metabolites is separated by thin-layer chromatography. The primary aromatic amines are diazotized and coupled with N-(1-naphthyl) ethylene-diamine on the thin-layer plate. The quantities of the 7-amino-metabolites are directly evaluated by colorimetric densitometry. The lower limit of detection is in order of 0.5–10 ng cm^?3 of plasma. The relative standard deviation of the whole procedure is less than ±15% in the range of 0.5–10 ng cm^?3 for a single determination. The unchanged drugs can be reduced on the thin-layer plate and detected by the same method. Since this procedure is rather difficult to perform, it is advantageous to determine the 7-nitro-benzodiazepines in plasma by gas chromatography. The thin-layer chromatographic method was used to measure the 7-amino-metabolites in plasma of patients on either a flunitrazepam or a clonazepam oral dosing regimen.     

Assay of tinidazole in human serum by high-performance thin-layer chromatography--comparison with high-performance liquid chromatography

Determination of tinidazole in human serum by high-performance thin-layer chromatography (HPTLC) is presented. It includes use of 10 x 10 cm plates coated with silica gel 60 and chloroform-acetonitrile-acetic acid (60 + 40 + 2) as mobile phase. Quantitation was performed by densitometry at 320 nm. The linearity (1-10 ng), precision (6%), reproducibility (5%), recovery (96%), and detection limit (1 mg/L) of tinidazole determination by HPTLC were comparable with corresponding method parameters by reversed-phase HPLC. A satisfactory correlation was found between the 2 analytical methods. The procedure was used to quantitate tinidazole in patient sera.     

Simultaneous determination of trace of loxacin, ciprofloxacin, and sparfloxacin by micelle TLC-fluorimetry

The method of simultaneous determination of ofloxacin (OFLX), ciproflxacin (CPLX), and sparfloxacin (SPLX) by thin-layer chromatography is established, with micelle solutions as mobile phases. It is found that the optimum molar ratio of sodium dodecyl sulfate (SDS) to ethylene diamine tetraacetic acid is 0.01:0.1. On the polyamide thin-layer sheet, OFLX, CPLX, and SPLX are separated from each other, and the corresponding Rf values are 0.72, 0.55, and 0.32, respectively. The fluorescence spots are scanned with a spectrodensitometer at the excitation wavelength of 282 nm. The cut-off filter is set at 400 nm. The detection limits are 2 x 10(-6) mol/L for OFLX, 1.5 x 10(-6) mol/L for CPLX, and 1.6 x 10(-6) mol/L for SPLX, and the respective linear ranges correspondingly fell in the concentration of 1 x 10(-5) to 4 x 10(-4) mol/L for OFLX, 1 x 10(-5) to 4.5 x 10(-4) mol/L for CPLX, and 1 x 10(-5) to 4.2 x 10(-4) mol/L for SPLX. For all the three components, the relative standard deviations are in the range of 1.12-5.82%, and the recoveries are found to be 96.7-104.2% in urine and serum samples.     

Determination of the serum concentration of spironolactone and its metabolites by high-performance liquid chromatography

A simple and rapid high-performance liquid chromatographic assay is described for the simultaneous determination in serum of the aldosterone antagonist spironolactone and its metabolites 7 alpha-thiomethylspirolactone, 6 beta-hydroxy-7 alpha-thiomethylspirolactone and canrenone. Ultraviolet absorption at 240 nm was used to detect the different compounds after elution on a normal-phase column. Endogenous serum substances did not interfere with the assay. This method provides a convenient tool in pharmacokinetic studies of spironolactone, in contrast to previously reported aspecific fluorimetric assays or time-consuming thin-layer chromatographic analyses of radioactive biological material.     

Adsorption of bovine serum albumin by granulated and fibrous sorbents

Adsorption of bovine serum albumin by macroporous glasses and fiber sorbents was studied. Differences in the mechanism of protein Sorption by sorbents depending on the chemical nature of the surface and the texture of the initial matrix were found. Optimum conditions for protein immobilization on thin-layer sorbents were determined. The reversibility of protein adsorption was studied by the radioactive indicator technique. The extent of reversibility of protein adsorption by thin-layer titanium hydroxide coatings was shown to depend significantly on the texture of the initial matrix.Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 8, pp. 1586–1589, August, 1995.     

Thin-layer chromatographic fingerprinting of the nonvolatile fraction extracted from the medicinal herb Cistus incanus L.

ABSTRACT

The aim of this study was to provide the first from-start-to-end thin-layer chromatographic method of fingerprinting the Cistus incanus L. raw herbal material, with a purpose to further use it for rapid screening, authentication, and quality control of the traded C. incanus L. herbs. To this effect, 12 different C. incanus L. samples purchased as herbal teas from a local market were extracted by means of the accelerated solvent extraction (ASE) with chemometrically optimized solvent extraction mixture and temperature (methanol–water, 27:73, v/v; 130°C), to derive the polar fraction from the plant samples. Then, the extracts were developed in two thin-layer chromatographic systems, both using the commercially precoated silica gel 60 chromatographic plates, yet two different mobile phases (mobile phase 1, ethyl acetate–formic acid–acetic acid–water, 100:11:11:13, v/v/v/v, and mobile phase 2, ethyl acetate–dichloromethane–formic acid–acetic acid–water, 100:10:10:10:11, v/v/v/v/v). The chromatograms were densitometrically scanned in the reflectance mode at the wavelength λ?=?366?nm to obtain fingerprints of the extracts derived from individual C. incanus L. samples. Mobile phase 2 performed slightly better, because with its use, the maximum number of 11 peaks could be seen in the respective fingerprints, whereas with mobile phase 1, the maximum number of 10 peaks only. Then an antioxidant potential of the investigated herbal extracts was assessed, making use of mobile phase 2 and the 0.20% methanol solution of 2,2-diphenyl picrylhydrazyl as a visualizing reagent. The resulting chromatograms were densitometrically scanned in the extinction mode at the wavelength λ?=?550?nm to obtain biological fingerprints of the extracts. Finally, chromatographic and biological fingerprints underwent a semiquantitative evaluation in terms of the contents of the extracted polar fraction and an overall antioxidant potential of the individual plant species.     

Evaluation of a high-performance thin-layer chromatographic technique for the determination of salbutamol serum levels in clinical trials.

Salbutamol concentrations were determined by high-performance thin-layer chromatography in the sera of two sets of ten volunteers at hourly intervals for 6 h after taking one 8-mg slow-release tablet. The influence of time lapse in processing of serum samples, i.e. centrifugation, extraction and chromatography, was studied. A statistical significant instability of salbutamol in the sera of patients was found which was not present in standard drug-free serum samples spiked with salbutamol and used for construction of standard curves.     

Amperometric lactate biosensor for flow injection analysis based on a screen-printed carbon electrode containing Meldola's Blue-Reinecke salt, coated with lactate dehydrogenase and NAD

A biosensor for the measurement of lactate in serum has been developed, which is based on a screen-printed carbon electrode, modified with Meldola's Blue-Reinecke Salt (MBRS-SPCE), coated with the enzyme lactate dehydrogenase NAD⁺ dependent (from Porcine heart), and NAD⁺. A cellulose acetate layer was deposited on the top of the device to act as a permselective membrane. The biosensor was incorporated into a commercially available, thin-layer, amperometric flow cell operated at a potential of only +0.05 V vs. Ag/AgCl. The mobile phase consisted of 0.2 M phosphate buffer pH 10 containing 0.1 M potassium chloride solution; a flow rate of 0.8 ml min⁻¹ was used throughout the investigation. The biosensor response was linear over the range 0.55-10 mM lactate; the former represents the detection limit. The precision of the system was determined by carrying out 10 repeat injections of 10 mM l(+)lactic acid standard; the calculated coefficient of variation was 4.28%. It was demonstrated that this biosensor system could be applied to the direct measurement of lactate in serum without pre-treatment; therefore, this would allow high throughput-analysis, at low cost, for this clinically important analyte.     

Quantitative Determination of Propranolol in Human Serum by High-Performance Thin-Layer Chromatography

JPC – Journal of Planar Chromatography – Modern TLC - A high-performance thin-layer chromatography (HPTLC) method for the quantification of propranolol in human serum is developed and...     

A highly effective one-minute thin-layer chromatographic separation of unconjugated tri- and dihydroxy bile acids

A rapid, miniature thin-layer chromatographic procedure has been developed for the separation of unconjugated tri- and dihydroxy bile acids. The new solvent system consisted of isooctane:diisopropyl ether:methanol:acetic acid:formamide (2:1:0.13:0.07:0.02,$\text{v}\text{v}$). Complete separation of tri- and dihydroxy bile acid fractions was easily achieved within 1 min on ITLC type SG chromatography sheets. A sensitive fluorescent visualization technique was employed. The reflected fluorescence intensity of the bile acids fluorophore spots was measured with a Farrand Mark I spectrofluorometer equipped with a thin-layer scanning attachment. The excitation and emission wavelengths were 375 and 436 nm, respectively. The above procedure has been adapted for the separation of unconjugated bile acids from serum samples. The bile acids were adsorbed onto Amberlite XAD-2, eluted with ethanol, and concentrated. Further purification was performed by a simple 3-min thin-layer chromatographic technique. This new procedure for the separation of tri- and dihydroxy bile acids is rapid, sensitive, and less complex than conventional procedures presently employed in clinical laboratories.     

High-performance liquid chromatographic determination of cholesteryl esters in the blood of obese children.

The serum of obese children and adolescents was analyzed for cholesteryl esters. The test substances were first separated from the sample matrix by solvent extraction and thin-layer chromatography and then resolved in a reversed-phase high-performance liquid chromatographic system involving a Separon SGX C18 column and a mobile phase of 2-propanol-acetonitrile (40:60, v/v), with ultraviolet detection at 206 nm. Cholesterol and 10-cholesteryl esters could be separated and determined within ca. 25 min at a flow-rate of 1 ml/min. The method was applied to a study of the effect of external conditions (physical stress, diet) on the content of cholesteryl esters in a test group of obese boys and girls aged from 13 to 16 years. The analyses have demonstrated that the above conditions do not affect the concentrations of the individual cholesteryl esters, although the total cholesterol concentration decreased significantly after spa treatment.     

Determination of organophosphorus pesticides by thin-layer chromatography

Analysis for 13 common organophosphorus pesticides by thin-layer chromatography is described; 17 solvent systems were examined. With channel thin-layer chromatography, linear calibration graphs were obtained for the range 1-10 mug.