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The adulteration of herbal medicines by dexamethasone or prednisolone is regarded as a serious problem in many communities. Herein, a novel platform for the separation and quantification of both target steroids in herbal medicines based on electrochemical paper-based analytical devices (ePADs) has been created. The ePAD was composed of Whatman SG81 chromatography paper, 3D-printed devices and a commercial screen-printed electrode. Whatman SG81 silica-coated paper was used for the separation of dexamethasone and prednisolone based on the difference in their partition coefficients during the flow of the mobile phase. The optimal mobile phase was composed of 60% ethyl acetate in cyclohexane and required 7 min for separation. The separated steroids on the paper were then quantified by electrochemical detection using differential pulse voltammetry, in which the 3D-printed devices facilitated the measurement. Analytical detection ranges of 10–500 μg mL−1 were obtained for both dexamethasone and prednisolone (r2 = 0.988 and 0.994, respectively). The limits of detection for dexamethasone and prednisolone were 3.59 and 11.98 μg mL−1, respectively, whereas the limits of quantification were 6.00 and 20.02 μg mL−1, respectively. The amounts of both target steroids derived from real herbal medicine samples determined by the proposed method were comparable to those obtained with assays using standard high-performance liquid chromatography. In addition, a simple evaporation step can be used to increase the concentration of the samples before analysis. These ePADs are simple, low-cost, rapid, and very promising for on-site quantitative detection.

This paper describes the development of two microfluidic paper-based analytical devices (μPADs), one well-based and the other based on a lateral flow assay (LFA) configuration, to detect glucose via a colorimetric assay using the solid metal-organic framework (MOF) Zr-PCN-222(Fe), to encapsulate glucose oxidase (GOx). The well-based platform consisted of laminate sheets and multiple layers of wax-printed chromatography paper. Solutions of KI and glucose placed into the well flowed through the device and reacted with the GOx@MOF species sandwiched between the paper layers realizing a yellow-brown color. The LFA platform consisted of chromatography paper between parafilm and polyvinyl acetate (PVA) layers. GOx@MOFs spotted on the paper subjected to solutions of KI and glucose yielded a brown color. The devices were then dried, scanned, and analyzed yielding a correlation between average inverse yellow intensity and glucose concentrations. The development of these devices employing MOFs as biomimetic catalysts should further expand the applications of microfluidic technologies for sensors a variety of analytes.

Cellulose is an important component of green plants cell wall and other organisms. Cellulose from plants is mainly used to produce paper. Because of the widespread use of paper throughout the world, it causes the use of wood in large quantities that involves cutting down trees which can lead to negative impacts on ecosystems. Many research studied to find other raw materials that could replace wood and some focused on coconut coir because it contains high amount of cellulose which is enough to use in manufacturing. Coconut pulp is also the residue from coconut. It has 59 % cellulose. This makes it possible to use coconut pulp in paper industry. Paper for chromatography uses fiber from wood as an important component too. So, the purpose of this study was to identify whether cellulose from coconut pulp can replace the original material of chromatography paper. The procedure started from extracted impurity from coconut pulp and used the remaining for making chromatographic paper. Washed coconut pulp and removed residue coconut milk. Boiled clean coconut pulp for 10 minutes and dried in oven at 60 degree Celsius. Remove the residue fat with hexane. Drained hexane and dried in oven at 60 degree Celsius. Extracted cellulose by the soda process. Cellulose yield was 50.85%. Used extracted cellulose as raw material for making chromatographic paper. Tried different amount of cellulose per area of sieve to find the suitable for making paper. The appropriate paper area per extracted cellulose weight was about 81.25 cm2/g. Then trailed the paper used as stationary phase for color separation. The appropriate condition of mobile phase in chromatographic technique (ethyl-methyl ketone, acetone and water) to separate standard colors was also studied. The appropriate ratio of mobile phase which could separate standard solutions was 7:3:3, respectively. The results show that this paper was effective in serving as a stationary phase. This paper has the potential to be developed to use as a substitute material in Bio manufacturing.

A colorimetric probe N,N’-bis(2-methoxy-ethyl)-2,3,3-trimethyl-3H-squaraine (MOESQ) with H2O solubility was synthesized to detect Cu2+. MOESQ exhibits good selectivity, high sensitivity and fast UV-Vis response toward Cu2+ over other competing ions in CH3CN. The detection limit of MOESQ for Cu2+ in CH3CN can reach 1.88 × 10−7 molL−1. By adsorbing MOESQ on the chromatography paper, a colorimetric test paper for Cu2+ was prepared, which could detect Cu2+ with the color change from blue to faint yellow even in the limit of detection concentration of 10−6 molL−1.

In this paper, we demonstrate the fabrication of gel beads composed of ionic liquid (IL) and polymer. The IL droplets are kept spherical during the deposition process by placement onto chromatography paper coated with fluoropolymer. The deposition process then occurs in two steps. In the first step, the monomer is absorbed into the IL droplet. In the second step, the initiator radicals are introduced. This sequential deposition process allows polymerization to primarily occur within the liquid droplet and therefore the beads are not attached to the underlying substrate and can be easily removed.

Vitex negundo L. is one of the most important species in traditional system of medicine to cure various ailments. Alkali soluble polysaccharide (ASP) from V. negundo stems was isolated and purified in 0.61% yield. Complete hydrolysis of ASP followed by paper chromatography and GLC analysis indicated the presence of l-rhamnose, l-arabinose, d-xylose, d-galactose and d-glucose in mole percent of 1.28, 2.25, 73.49, 8.08, 8.11 along with 2.48 d-galacturonic acid and 4.27 d-glucuronic acid. Uronic acids were also estimated by spectrophotometric methods using carbazole, m-hydroxydiphenyl and 3, 5-dimethyl phenol (DMP) as colorimetric reagents. Estimation of uronic acids using DMP corroborated the results of GLC analysis. The study evaluated the utility of colorimetric methods for uronic acid estimation in polysaccharide having interferences from neutral sugars especially xylose, glucose and galactose. The analysis also resulted in composition of constituent monosaccharides of ASP and co-relation analysis of uronic acids content. The results may divulge the structural moiety of the polysaccharide responsible for its bio-efficacy.

Although paper chromatography is being promoted as a cost-effective tool for rapid assessment of soil health, few studies have explored the quantitative relationship between chromatogram features and soil health variables, and no studies have investigated the association between chromatogram features and microbial diversity as determined using DNA sequences. We assessed 343 soil samples from varying land uses in southwestern Australia to investigate the relationship between total organic carbon, microbial activity and diversity, salinity and pH levels and 12 chromatogram features. Spearman’s correlations and variance partitioning were used to detect relationships. Although total organic carbon and microbial activity displayed the strongest correlations with chromatogram features, they were not associated with greater development of radial features and median zone radius as expected. This was in contrast to what has been previously reported, implying context dependent responses of chromatogram features to gradients in soil variables. Microbial community structure was found to explain changes in chromatogram features better than the measured soil variables. These results indicate that further studies are necessary before paper chromatography is embraced as a tool for soil health assessment, and raises questions regarding the use of chromatography by community groups as a tool to measure soil health.

High-performance affinity chromatography is limited by its high cost and high pressure. Paper is made up of porous fiber networks and has the properties of low cost, ease of fabrication, and biodegradable. Due to these advantages, herein, we immobilized beta2-adrenoceptor (β2-AR) onto the surface of the polytetrafluoroethylene membrane, a paper-based material, and constructed a G protein-coupled receptor (GPCR)-in-paper chromatographic platform. This platform was characterized by Fourier transform infrared spectroscopy, fluorescence analysis, X-ray photoelectron spectroscopy, and chromatographic studies. These morphological and elemental analysis showed that β2-AR was successfully immobilized on the paper surface. The specific drugs have good retentions on the GPCR-in-paper chromatographic platform. The association constants of salbutamol, terbutaline and bambuterol to β2-AR were calculated to be 2.02 × 104 M−1, 1.15 × 104 M−1, 1.75 × 104 M−1 by adsorption energy distribution, which were in good line with the values from frontal analysis, zonal elution and previous literatures. We demonstrated that the GPCR-in-paper platform was cost-effective, easy to be modified for protein immobilization, and applicable in the receptor-drug interaction analysis. We believe such a platform sheds new light on paper chromatography for receptor-drug interaction analysis and other applications.

Recently, the anticancer activity of ferulic acid (FA) which is a caffeic acid derivative has been reported. Therefore, in this study FA was radiocaped with 131I− to explore its potential as a tumor targeting agent. The radiolabeling process was carried out via electrophilic substitution reaction. The factors affecting labeling yield were optimized and the radiochemical purity (RCP) was assessed by various analytical techniques including paper chromatography (PC), thin layer chromatography (TLC), instant thin layer chromatography (ITLC), paper electrophoresis (PE) and high-performance liquid chromatography (HPLC). The RCP assay was extended to the utilization of miniaturized techniques including miniaturized PC (mini-PC), mini-TLC and mini-column chromatography (silica, sephadex-G25). Validation of mini-TLC, as one of 131I-FA RCP assay methods, was done according to ICH guidelines. Biodistribution studies of 131I-FA were performed on Ehrlich solid tumor bearing mice at various time points (5, 30, 60, 120 and 240 min), post injection. The radiolabeling yield of 131I-FA was 96.23 ± 0.45% and the miniaturized chromatographic systems showed high efficacy in RCP evaluation comparable to the conventional ones. Mini-TLC was proved to be specific, accurate, precise and linear. The tumor uptake of 131I-FA in solid tumor bearing mice was 4.35 ± 0.41 ID/g at 60 min with 2.79 as a tumor/muscle ratio. Consequently, 131I-FA could be used as a tumor targeting agent for nuclear medicine applications and the fast reaction monitoring could be achieved using miniaturized chromatographic techniques.

The objective of this work was to develop a simple microfluidic paper-based analytical device (µPAD) for the determination of paraquat. The µPAD was simply and rapidly made by dipping chromatography paper in melted paraffin. It was observed that the contact angle of the hydrophobic channel was 105.3 ± 4.4°. The analysis of paraquat was based on the reaction of paraquat in basic medium with sodium dithionite, giving the blue radical ions on the test zone. Measurement was made by taking photographs and measuring the red color intensity using the ImageJ program. Under the optimized conditions, the working range was determined to be 1.24–20.00 mg/L, LOD and LOQ were 1.24 and 4.13 mg/L with RSD for intraday and interday precision of 8.28% and 17.00%, respectively. The proposed µPAD device was applied for paraquat determination in several types of vegetable samples giving recovery of 89.9–104.9%. The results were not significantly different from spectrophotometry at the 95% confidence level.

Herein, we report three-dimensional paper chromatography (3D-PC) as a micro-chromatographic platform. The method was based on applying the origami microfluidic device for separation, coupled by colorimetric methods for simultaneous determination. The microfluidic device fabrication was a facile printing approach. Two azo food dyes, Tartrazine (E102) and Indigo carmine (E132), were selected as a model analyte, while carbonate-bicarbonate buffer was used as the mobile phase. Our micro-chromatographic device is associated with two big advantages including needing very small volume of mobile phase ( ~12 µL) and ultrafast separation time (~35 s). Under the optimal conditions, the method provided acceptable linear ranges of 0. 0 g L 1–18.0 g L 1 (R2 = 0.997) for E102 and 0.070 g L 1–10.0 g L 1 for E132 and the limits of detection (3σ/slope) were evaluated as 0.620 and 0.060 g L 1, respectively. The proposed method was successfully applied in the separation and quantification of these dyes in commercial food products such as jelly, candy, and four kinds of drink samples without any sample preparation prior to analysis. The mean recovery values for the real sample analysis were in the range of 100.14%–102.38% for E132 and E102 respectively. The inter-device relative standard deviations were in the ranges of 1.5%–11.8%. In total, our chromatographic μPAD is small (1.0 cm × 1.0 cm × 0.5 cm), portable, inexpensive, no need of specialized user, requires low volumes of sample (0.5 μL), and can perform separation using 12 μL of aqueous mobile phase in very short time.

A facile and simple paper-based scanometric assay was developed to detect Pb(2+) using GR5-DNAzyme. Magnetic beads (MBs) and gold nanoparticles (AuNPs) were used as a signal collector and a signal indicator, respectively. They were linked together by GR5-DNAzyme, comprising an enzyme and a substrate strand pairing up with each other. In the presence of Pb(2+), the substrate strand is cut into two pieces, resulting in the disassembly of AuNPs from the MBs. These AuNPs were spotted on predefined areas on a chromatography paper, where signal is amplified through silver reduction. This sensing platform exhibits high sensitivity and selectivity toward Pb(2+), giving a detection limit of 0.3 nM and a linear fitting range from 0.1 to 1000 nM. Testing of this biosensor in river water and synthetic urine samples also showed satisfying results. Besides offering simultaneous and multi-sample analysis, this paper-based sensing platform presented here could be potentially applied and served as a general platform for on-site, naked eyes, and low-cost monitoring of other heavy metal ions in environmental and body fluid samples.

Two series of “papers” that were made from natural fibers and synthetic fibers, respectively, were examined for use in paper-spray mass spectrometry and the results were compared to chromatography paper that is currently being used. In the former case, four types of papers were used, including gampi paper, tengujou paper, glassine paper and cicada paper, and the findings show that the limit of detection can be improved when gampi paper was used. This is because gampi paper is very tough and extremely thin (thickness, l -lactic acid (PLLA), respectively, by means of a co-axial electrospinning technique. The findings show that the limit of detection also can be improved, when a PLLA nanofiber was used. This is because this type of paper-like nanofiber is also very thin, tough and hydrophobic, which permits to ionization to occur within a very short period. Detailed information on methods for synthesizing these fibers and their use in the analysis of a real sample are also reported.

This Letter aimed to develop an efficient method for the determination of cyanide ion (CN − ). A novel colorimetric chemosensor 4-[(1 E )-2-(4-hydroxyphenyl)ethenyl]-1-allylpyridinium bromide (HPEAPB) was synthesized. HPEAPB displayed good selectivity toward CN − over other competing anions in ethanol. A color change from yellow to red was immediately observed upon the addition of CN − and the limit of detection (LOD) was 3.4 × 10 −6 mol L −1 . The sensing mechanism was discussed by UV–vis, 1 H NMR titration, and a comparison study. Colorimetric test paper for CN − was prepared by attaching HPEAPB to a chromatography paper, which could be used to detect CN − in environmental samples as simply as a pH-indicator paper for pH value. The LOD of the test paper for CN − was 2.0 × 10 −4 mol L −1 . This detection method for CN − has potential applications in cyanide ion containing fields by combination of rapid and real-time advantages.

The anthocyanins in the brilliant crimson petals from Erythrina crista-galli L. were extracted with 10% AcOH, and the extract was purified by Amberlite XAD-7 column chromatography, paper chromatography, and preparative HPLC. The structure of a major anthocyanin was confirmed as cyanidin 3- O -sophoroside by 1 H and 13 C NMR and FABMS. Then, the catalysis of UDP-glucose: flavonoid 3- O -glucoside glucosyltransferase (3GGT) was predicted at the final step in the anthocyanin biosynthesis. In this study, the 3GGT was purified approximately to 800-fold from E. crista-galli petals by eight-steps purification procedures. The purified enzyme (Ec3GGT) catalyzed only the 2″- O -glucosylation of flavonoid 3- O -glucoside to produce flavonoid 3- O -sophoroside. Ec3GGT showed a pH optimum around 7.5. The molecular weight and isoelectric point were estimated to be about 82 kDa and 6.5, respectively; the molecular weight was larger than known UDP-sugar: flavonoid glycosyltransferases (UFGTs). The enzyme did not require metal cofactors but inhibited by Zn 2+ , Co 2+ and Cu 2+ , and it activity was inhibited by SH-blocking reagents such as NEM and PCMB. Both anthocyanidin 3- O -glucosides and flavonol 3- O -glucosides were favorable acceptor substrates of the enzyme, whereas no activity was shown toward C -glucosylflavones such as isovitexin, which is a minor component in the petals. These results showed that Ec3GGT involved in the last step of biosynthesis of a major anthocyanin in petals.

Cold water-soluble (CWSP) and hot water soluble polysaccharides (HWSP) from Tinospora cordifolia stems were isolated and purified in 2.99% and 1.99% yield respectively. Complete hydrolysis followed by paper chromatography and GLC analysis indicated the presence of L-rhamnose, L-arabinose, D-xylose, D-mannose, D-galactose and D-glucose in molar ratio of 0.857, 1.106, 0.727, 0.526, 0.708 and 95.763 in CWSP and 0.697, 0.777, 2.048, 0.777, 0.292 and 95.408 in HWSP. The uronic acid content in the polysaccharide has been studied extensively using assorted approaches. It was quantitatively estimated by GLC analysis and spectrophotometric methods using carbazole, m-hydroxydiphenyl and 3,5-dimethylphenol as colorimetric reagents. GLC analyses indicated galacturonic acid content of 3.06% and 5.16% in CWSP and HWSP respectively. Estimation of uronic acid using 3,5-dimethylphenol corroborated the above analysis. The study resulted in composition of constituent monosugars of CWSP and HWSP and co-relation analysis of uronic acid content, leading to an unambiguous structural analysis.

Introduction Since Pall-Gelman stopped manufacturing ITLC-SG, it has become necessary to validate alternative stationary phases. Objective To validate different stationary phases versus ITLC-SG Pall-Gelman in the determination of the radiochemical purity (RCP) of 111 In-pentetreotide ( 111 In-Octreoscan) by planar chromatography. Material and methods We conducted a case–control study, which included 66 111 In-pentetreotide preparations. We determined the RCP by planar chromatography, using a freshly prepared solution of 0.1 M sodium citrate (pH 5) and the following stationary phases: ITLC-SG (Pall-Gelman) (reference method), iTLC-SG (Varian), HPTLC silica gel 60 (Merck), Whatman 1, Whatman 3MM and Whatman 17. For each of the methods, we calculated: PRQ, relative front values ( R F ) of the radiopharmaceutical and free 111 In, chromatographic development time, resolution between peaks. We compared the results obtained with the reference method. The statistical analysis was performed using the SPSS program. The p value was calculated for the study of statistical significance. Results The highest resolution is obtained with HPTLC silica gel 60 (Merck). However, the chromatographic development time is too long (mean = 33.62 min). Greater resolution is obtained with iTLC-SG (Varian) than with the reference method, with lower chromatographic development time (mean = 3.61 min). Very low resolutions are obtained with Whatman paper, essentially with Whatman 1 and 3MM. Therefore, we do not recommend their use. Conclusions Although iTLC-SG (Varian) and HPTLC silica gel 60 (Merck) are suitable alternatives to ITLC-SG (Pall-Gelman) in determining the RCP of 111 In-pentetreotide, iTLC-SG (Varian) is the method of choice due to its lower chromatographic development time.

This paper describes a convenient procedure for the radiochemical preparation of d -[U- 14 C]-glucose, d -[U- 14 C]-fructose and [U- 14 C]-sucrose with high specific activity by photosynthesis using ‘canna indica’ leaf, [ 14 C]-carbon dioxide and water in presence of light in a closed system. The [ 14 C]-sugars formed were extracted, separated and then purified by paper chromatography. Further, the pure d -[U- 14 C]-glucose obtained was converted to methyl-α- d -glucopyranoside ([U- 14 C]-glucose) by glycosidation with methanol using (i) HCl, the conventional Fischer method (ii) heterogeneous organic cation exchange resin (Amberlite IR-120 (H + )) and (iii) heterogeneous inorganic cation exchanged montmorillonites called metal M +n -monts. The results indicated that the latter in the form of Fe +3 -montmorillonite gave a better yield ( 65%) as compared to others (40–56%). The radiochemical purity of the no-carrier added product was more than 98%. The product retained its specific activity as that of the starting material which is in the range of 250–300 mCi/mmole (9.25–11.1 GBq/mmole), suitable for use as a radiotracer in biochemical investigations.

Aqueous normal-phase chromatography utilizing silica-hydride-based stationary phases provides unique selectivities for the separation of polar and non-polar compounds and offers new avenues to solve difficult analytical separations. By varying the amount of water in the mobile phase, all silica-hydride-based separation materials can operate in the reversed-phase mode, the normal-phase mode or, in some instances, a combination of both. This review describes some of the fundamental properties of silica-hydride materials and applications that encompass a broad range of separation challenges. Moreover, exemplars are provided that illustrate many of the prominent features that lead to the selection of such stationary phases for solving a particular analytical problem.

An overview is presented of the impact of chromatographic method developments on the quality control of pharmaceuticals as of the 1950s up until the present times. This survey is mainly based on the changes in pharmacopeias starting with United States Pharmacopeia (USP) 16, issued in 1960, up to the presently effective USP 34 and European Pharmacopeia 7.2. At the beginning of this period the role of chromatographic methods was negligible and was restricted to classical column chromatography and paper chromatography. However, the invention of high-performance liquid chromatography (HPLC) initiated a rapid paradigm shift in the attitude toward, and the use of, chromatographic methods. As a result, HPLC began a “career” of rapid spreading and development, and by now has undoubtedly become the principal method in pharmaceutical analysis. Likewise, the role of thin-layer chromatography (TLC) and to a lesser extent gas chromatography is also remarkable. The role of these and electrophoretic methods in the identification, assay and purity check of drugs and drug products in the modern pharmacopeias is discussed. As a case study the stability investigation of Depersolone ® injection carried out in the 1960s and 35 years later is presented and the information obtainable from the classical and modern approach is compared.

The selection of the mobile and the stationary phase in support-free liquid–liquid chromatography (centrifugal partition chromatography and counter-current chromatography) is equivalent to a selection of a biphasic liquid system and its global composition. There is an immense number of choices of biphasic liquid systems. On one hand what makes this technique extremely versatile, on the other hand turns the selection of the appropriate system for a particular separation problem into a challenging and demanding task. In this work a systematic procedure for the selection of biphasic liquid systems for preparative scale separations is presented. The procedure is adaptable to the production scale requirements including production cost and safety. The experimental effort of different stages of the selection procedure is minimized by using a fully predictive method, the conductor-like screening model for real solvents (COSMO-RS). The COSMO-RS is used to assess properties relevant for the selection of a biphasic liquid system, such as the solute solubility and the partition coefficient.

We developed a convenient method by coupling frontal elution paper chromatography with desorption corona beam ionization mass spectrometry (DCBI-MS) for rapid determination of chlorphenamine added in herbal medicines or dietary supplements. In this method, the ethanol extract of the herbal products was spotted directly onto an isosceles triangular filter paper sheet, and then the paper sheet was developed under strong elution condition with the sample zone migrating at the solvent front. The analyte was finally condensed at the V-shaped tip which could then be placed under the visible plasma beam of DCBI for ionization. The overall procedure took less than 5 min. The frontal elution paper chromatography on a triangular plate used in this work improved the signal intensity of chlorphenamine by 30-fold due to the analyte condensing at the tip and the reduction of the background suppression. Furthermore, the paper sheet also functioned as a filter in the analysis of solid or powder samples, which can increase the analytical throughput by omitting the step of centrifugation. The proposed method in current study was successfully applied in the determination of chlorphenamine in herbal medicines. Chlorphenamine was detected in four of the twelve types of herbal medicines examined in this study. The limit of detection was 200 ng/mL (2.0 ng absolute) in full-scan positive-ion mode and the linear range was from 5.0 μg/mL to 50 μg/mL with satisfactory linear coefficient (R(2) (the square of the correlation coefficient)=0.895). Good reproducibility was achieved with relative standard deviations (RSDs) less than 15.0% and the recoveries of chlorphenamine ranged from 84.3 to 90.6%.

Experiments have been carried out to evaluate Counter Current Chromatography (CCC) as an alternative purification technique to preparative Reverse Phase High Performance Liquid Chromatography (RP-HPLC) for small molecule pharmaceuticals. The major drawback of CCC is the extensive time required in selecting the solvents to perform the separation. This is equivalent to choosing both the stationary phase and the mobile phase at the same time. In RP-HPLC it is a simple matter of deciding on the gradient, most samples can be purified on a C18 column with a water:acetonitrile gradient. The majority of the initial work was based on a standard test set of commercially available compounds, developed within our group to evaluate the performance of the HPLC apparatus and the column prior to the start of work each day. The work carried out on CCC has shown that the technique offers similar capabilities and can be carried out using similar protocols to RP-HPLC. CCC also has some advantages over RP-HPLC and can be regarded as a valuable addition to the chromatography toolbox.

Bacterial colonies are spatially complex structures whose physiology is profoundly dependent on interactions between cells and with the underlying semi-solid substratum. Here, we use bacterial colonies as a model of a microbial community to evaluate the potential of laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) to delineate elemental distributions within colonies with minimal pre-treatment. To reduce water content of the colony and limit undesirable absorption of laser energy, we compared methods of preparing 24h-old colonies of Escherichia coli TG1 on agar for laser ablation. Colonies on excised agar segments dried on chromatography paper were superior to colonies dried in a dessicator or by prolonged incubation, with respect to signal magnitude, signal:noise ratio and background signal. Having optimised laser scan speed (10 microm s(-1)) and laser beam diameter (100 microm), further improvements were achieved by growing colonies on nylon membranes over agar, which were then transferred to the ablation chamber without further treatment. Repeated line rasters across individual membrane-supported colonies yielded three-dimensional elemental maps of colonies, revealing a convex morphology consistent with visual inspection. By normalising isotope counts for P, Mn, Zn, Fe and Ca against Mg, the most abundant cellular divalent cation, we sought elemental heterogeneity within the colony. The normalised concentration of Mn in the perimeter was higher than in the colony interior, whereas the converse was true for Ca. LA-ICP-MS is a novel and powerful method for probing elemental composition and organisation within microbial communities and should find numerous applications in, for example, biofilm studies.

Dopamine–HCl and l -DOPA-α-glycosides were prepared by reaction with cyclomaltohexaose, catalyzed by Bacillus macerans cyclomaltodextrin glucanyltransferase. The reaction gave maltodextrins attached to dopamine and l -DOPA; the maltodextrins were trimmed by reactions with glucoamylase and β-amylase to produce α-glucosyl- and α-maltosyl-glycosides, respectively. The glucoamylase- or β-amylase-treated dopamine- and l -DOPA-α-glycosides were fractionated and purified by BioGel P-2 gel-filtration column chromatography and preparative descending paper chromatography. Analysis by MALDI-TOF mass spectrometry and one- and two-dimensional NMR showed that the purified glycosides of dopamine and l -DOPA were glycosylated at the hydroxyl groups of positions 3 and 4 of the catechol ring. The major product was found to be 4-O-α-glycopyranosyl l -DOPA, and it was shown to be more resistant to oxidative tolerance experiments, involving hydrogen peroxide and ferrous ion, than l -DOPA. l -DOPA-α-glycosides are possibly more effective substitutes for l -DOPA in treating Parkinson’s disease in that they are more resistant to oxidation and methylation, which renders l -DOPA ineffective and deleterious.

A water-soluble polysaccharide isolated from Dalbergia sissoo Roxb. leaves was purified and major homogeneous fraction obtained by GPC. Complete hydrolysis of the polysaccharide followed by paper chromatography and GLC analysis indicated the presence of l -rhamnose, d -glucuronic acid, d -galactose and d -glucose in molar ratio of 1:1:2:2.33, respectively. Partial hydrolysis of the polysaccharide furnished one tri-[I], one hepta-[II] and one nona-[III] saccharides. Hydrolysis of the oligosaccharide I, II and III followed by GLC analysis furnished d -glucose and l -rhamnose (2:1); l -rhamnose, d -galactose and d -glucuronic acid (1:3:3); and l -rhamnose, d -galactose and d -glucose (1:3:5), respectively. Methylation analysis and periodate oxidation of the oligosaccharide I indicated the presence of two non reducing glucose units linked to rhamnose by 1→2 and 1→4 linkages, respectively. Oligosaccharide II is a branched molecule with a main chain consisting of 1,3-linked β- d -galactopyranosyl (2 mol), 1,3,4 linked α- l -rhamnopyranosyl (1 mol) and 1,4,6 linked β- d -galactopyranosyl unit (1 mol) and non reducing β- d -glucuronic acid at the end along with side chains of β- d -glucouronopyranosyl units (2 mol). Oligosaccharide III is also a branched molecule with a main chain consisting of 1,3,4 linked α- l -rhamnopyranosyl (1 mol), 1,2,4 linked β- d -glucopyranosyl (1 mol), 1,3 and 1,4 linked β- d -galactopyranosyl (2 and 1 mol, respectively) having β- d -glucopyranosyl as a non reducing end.

IPS-B2, an intracellular polysaccharide with anti-tumor effect, was isolated from cultured Armillariella tabescens mycelia by hot water extraction, anion-exchange and gel chromatography. Based on the results of paper chromatography (PC), gas chromatography (GC), infra-red (IR) spectroscopy and (13)C NMR, IPS-B2 was characterized as an α-(1→6)-d-glucan with a molecular weight of 49.5kDa. The effects of IPS-B2 on murine peritoneal macrophages were further investigated. The results demonstrated that IPS-B2 induced nitric oxide (NO) and cytokines (TNF-α, IL-1β and IL-6) production in macrophages. The outcomes of real-time reverse transcription-polymerase chain reaction (RT-PCR) proved that the transcribing level of inducible NO synthase (iNOS), TNF-α, IL-1β and IL-6 mRNA in the peritoneal macrophages have been augmented by IPS-B2. These data suggest that the anti-tumor activity of the polysaccharide from A. tabescens may due to activation of macrophage.

In the present paper, a new chromatographic method for the determination of acquired immune deficiency syndrome (AIDS) drugs in plasma samples is proposed. The method consists of solid-phase extraction for sample pretreatment and further chromatographic analysis. Drugs have been separated on a C(18) column using an elution gradient based on an increase in the acetonitrile percentage. Analytes have been detected spectrophotometrically at 240, 250, 260 and 280nm. Chromatographic conditions have been thoroughly optimized using experimental design and multicriteria functions. Analytical parameters of the method have been established for both synthetic and plasma samples. Limits of detection are around 5ngmL(-1) for reverse transcriptase inhibitors (nucleoside and non-nucleoside) and 20ngmL(-1) for protease inhibitors. The method has been validated through a spiking/recovery procedure at three concentration levels. Results obtained are highly satisfactory, with recovery values around 100% for all drugs. The method has been applied to the determination of various drug mixtures of medical interest in plasma samples.

The hexasaccharide ajugose, alpha-D-galactopyranosyl-(1--6)-alpha-D-galactopyranosyl-(1--6)-O-alpha-D-galactopyranosyl-(1--6)-alpha-D-galactopyranosyl-(1--6)-alpha-D-glucopyranosyl-(1--2)-beta-D-fructofuranoside, generally uncommon in legumes, was detected in the seeds of Vigna mungo L. by TLC and paper chromatography. Ajugose was then isolated by silica gel chromatography and its structure was established by acid and enzymatic hydrolysis, fast atom bombardment mass spectrometry and both one- and two-dimensional 1H and 13C NMR techniques.

The gene encoding β-glycosidase of the hyperthermophilic archaea Sulfolobus shibatae (SSG) was expressed in Escherichia coli . Recombinant SSG (referred to as rSSG hereafter) was efficiently purified, and its transglycosylation activity was tested with lactose as a donor and various sugars as acceptors. When sucrose was used as an acceptor, we found a distinct intermolecular transglycosylation product and confirmed its presence by TLC and high performance anion exchange chromatography (HPAEC). The sucrose transglycosylation product was isolated by paper chromatography, and its chemical structure was determined by 1 H and 13 C NMR. The sucrose transfer product was determined to be β- d -galactopyranosyl-(1→6)-α- d -glucopyranosyl-β- d -fructofuranoside with a galactose molecule linked to sucrose via a β-(1→6)-glycosidic bond.

Previous studies reached different conclusions about whether class I hyaluronan synthases (HASs) elongate hyaluronic acid (HA) by addition to the reducing or the nonreducing end. Here we used two strategies to determine the direction of HA synthesis by purified class I HASs from Streptococcus equisimilis and Streptococcus pyogenes. In the first strategy we used each of the two UDP-sugar substrates separately to pulse label either the beginning or the end of HA chains. We then quantified the relative rates of radioactive HA degradation by treatment with beta-glycosidases that act at the nonreducing end. The results with both purified HASs demonstrated that HA elongation occurred at the reducing end. In the second strategy, we used purified S. equisimilis HAS, UDP-glucuronic acid, and UDP[beta-32P]-Glc-NAc to radiolabel nascent HA chains. Under conditions of limiting substrate, the 32P-labeled products were separated from the substrates by paper chromatography and identified as HA-[32P]UDP saccharides based on their degradation by snake venom phosphodiesterase or hyaluronidase and by their binding to a specific HA-binding protein. The 32P radioactivity was chased (released) by incubation with unlabeled UDP-sugars, showing that the HA-UDP linkages turn over during HA biosynthesis. In contrast, HA-[32P]UDP products made by the purified class II Pasteurella multocida HAS were not released by adding unlabeled UDP-sugars, consistent with growth at the nonreducing end for this enzyme. The results demonstrate that the streptococcal class I HAS enzymes polymerize HA chains at the reducing end.

MraY is an established target for the discovery of antibacterial agents. The conventional assay for MraY uses radioactive sub- strate and analysis of products after paper chromatography or butanol extraction. Synthesis of radiolabeled substrate has been done in vitro using purified enzymes or by growing cells on radiolabeled precursors. The authors report a simple and rapid method to chemically radiolabel MraY substrate, UDP-MurNAc-pentapeptide. Specific activity obtained by this method was more than 100 times higher than the conventionally labeled substrate, and yields are high enough to support the requirements of high-throughput screening (HTS). The authors have developed a microplate-based homogeneous assay for MraY in which the product is captured on wheat germ agglutinin (WGA) scintillation proximity assay (SPA) beads. The assay was validated by showing inhibition by specific inhibitors of MraY but not by inhibitors of other enzymes of peptidoglycan synthesis. The assay uses wild-type membranes of Escherichia coli, giving it an advantage over recently described assays that need the pro- tein to be overexpressed. In addition, it has an advantage over the high-throughput MraY-MurG coupled assay reported in the literature because it is MraY specific, and therefore hits obtained in this assay do not need further deconvolution. It has potential for use in HTS approaches to find novel inhibitors of MraY. (Journal of Biomolecular Screening 2005:149-156)

The preparation of a polyazamacrocyclic-nitrotriazole conjugate for radiolabeling with the therapeutic radioisotope viz. (177)Lu is described. The nitroimidazole used for the present study is [N-2'(carboxyethyl)-2-(3'-nitro-1'-triazolyl)acetamide], the carboxylic acid derivative of sanazole, which possesses an optimal combination of desired properties such as, selective toxicity for hypoxic cells, lowered lipophilicity resulting in lowered neurotoxicity. The bifunctional chelating agent is a DOTA derivative viz. 1,4,7,10-tetraaza-1-(4'-aminobenzylacetamido)-cyclododecane-4,7,10- triacetic acid (p-amino-DOTA-anilide). (177)Lu was produced in adequate specific activity (110TBq/g) and high radionuclidic purity (approximately 100%) by irradiating enriched (60.6% (176)Lu) Lu(2)O(3) target and used for radiolabeling of the sanazole-BFCA conjugate. approximately 98% Complexation yield was achieved under optimized conditions. The complex has been characterized by paper chromatography and HPLC studies. Bioevaluation studies in Swiss mice bearing fibrosarcoma tumors revealed moderate tumor uptake (0.88%/g at 1h post-injection) with favorable tumor to blood (4.00 at 1h post-injection) and tumor to muscle (4.63 at 1h post-injection) ratios.

A method for the isolation of polysaccharides from the mucus of the loach, Misgurnus anguillicaudatus, has been devised. It involves the extraction of the skin mucus with tap-water (pH 7) at room temperature, followed by centrifugation, vacuum vaporization, Sevag extraction, precipitation with ethanol, vacuum filtration and lyophilization. The sediment preparation was identified to be a carbohydrate compound by several qualitative tests. Using Sephadex G-100 column (1×50 cm ) gel filtration to treat the sediment preparation, two major constituents, P and O, were yielded. P was identified to be a novel polysaccharide and named M. anguillicaudatus polysaccharide (MAP) or misgurnan. Its average molecular weight (Mw=1.30×105, Mn=1.23×105) was determined by gel permeation chromatography (GPC). It was characterized by UV, IR and NMR spectroscopy. The major structural monomers of MAP were identified to be d -galactose, l -fucose and d -mannose by paper chromatography (PC) and gas chromatography (GC). Smith degradation test showed that the monoses link each other by α-1,3 bonds. This is the first report on the chemical structure of a free neutral hetero-polysaccharide in loach mucus.

Walls were isolated from sheets of aleurone cells from wheat, freed from associated protein and starch by extraction with phenol-acetic acid-water followed by treatment with porcine pancreatic alpha -amylase, and finally extracted with sodium dodecyl sulphate. Hydroxycinnamic acids (HCA) released from the walls by saponification with 1 M NaOH amounted to 1·84% w/w of wall and comprised ferulic acid (90%), p coumaric acid (10%) and minor amounts (0·006%) of diferulic acids. The isolated walls exhibited an intense blue autofluorescence when irradiated with UV light that was reduced in intensity and became more diffuse after saponification. Acetyl residues (4·9% w/w) were present representing a degree of substitution of two in every nine pentose residues in the wall arabinoxylans. Digestion of the walls with a polysaccharide hydrolase mixture that lacked HCA esterases, released a series of oligomers that contained 50% of the total esterified HCA. These were separated by hydrophobic exclusion chromatography, purified by paper chromatography and two of these were identified by ES-MS and NMR as O -[5- O -feruloyl-α-L-arabinofuranosyl]-(1→3)- O -β-D-xylopyranosyl-(1→4)-β-D-xylopyranose (FAXX) andO -β-D-xylopyranosyl-(1→4)- O -[5- O -feruloyl-α-L-arabinofuranosyl]-(1→3)-O -β-D-xylopyranosyl-(1→4)-β-D-xylopyranose (XFAXX). The extremely low level of esterified diferulic acid indicates that cross-linking between polysaccharide chains by diester-FA is not important, however the esterified FA may play a role in cross-linking polysaccharides to wall proteins.

Driselase-digestion of cell walls from suspension-cultures of spinach (Spinacia oleracea L.), followed by anion-exchange chromatography, gel-permeation chromatography, preparative paper chromatography and preparative paper electrophoresis, yielded ten uronic acid-containing products in addition to free galacturonic acid (GalA). These included 4-O-methylglucuronic acid, alpha-L-rhamnopyranosyl-(1-->4)-D-glucuronic acid and several oligosaccharides containing GalA residues. The structures were unambiguously determined by a combination of 1- and 2-dimensional NMR spectroscopic techniques. Five of the six homogalacturonan-derived oligosaccharides purified contained 3-O-acetyl-GalA residues; however, methyl-esterified GalA residues occurred adjacent to both 2-O-acetyl-GalA and 3-O-acetyl-GalA residues. An acetylated, rhamnogalacturonan-I-derived oligosaccharide that was purified also contained 3-O-acetyl-GalA residues. Taken together with published data, our findings indicate considerable diversity in the patterns of pectin esterification. The implications for the action of pectin esterases are discussed.

The preparation of 166Ho labeled hydroxy apatite (HA) particles for radiosynovectomy applications is described in this paper. 166Ho was prepared by the irradiation of Ho2O3 at a flux of 1.8 x 10(13) neutrons/cm2/s for about 7 days. The irradiation resulted in the production of approximately 17 GBq of 166Ho activity at the end of six hours post end of bombardment and the corresponding specific activity was approximately 3-4 GBq/mg of Ho. The irradiated target was dissolved in 0.1 N HCl solution. Radionuclidic purity was ascertained by high resolution gamma ray spectrometry. HA particles were synthesized and characterized by X-ray diffractometry. Labeling studies were carried out with and without citric acid as a transchelating agent. Radiochemical yield and purity of the 166Ho-HA particles were ascertained by paper chromatography and by paper electrophoresis techniques. Labeling yield of >98% could be achieved at pH 7, with 40 mg of HA particles and 8.6 microg of Ho. 166Ho-HA particles prepared were stable for 72 h. Bio-evaluation of the 166Ho -HA particles were carried out by injecting approximately 74 MBq dose in 200 microL (approximately 8 mg of 166Ho-HA particles) directly into the arthritis induced knee joints as well as into the healthy knee joints of white New Zealand rabbits. Images of the injected joints of the animals recorded using a gamma camera at regular intervals showed good retention. Blood samples were collected from the animals and activity assayed in a scintillation detector. Experiments were also carried out under identical conditions in normal rabbits. In both the cases, it was observed that there was no significant extra articular leakage of the injected activity over the study period of 96 h post injection.

The crude protein of Zizyphus spina-christi seeds was found to be 15.9%. The percentage of essential amino acids was (12.8%) of total amino acids. Semi-essential was 17.1%, and non-essential represents 70% of the total mixture. Free sugars of the edible part of the fruit were isolated and identified by Paper Chromatography (PC) and High Performance Liquid Chromatograph (HPLC) and shown to be a mixture of fructose, xylose, glucose, and rhamnose. The yield of mucilage of the edible parts of the fruit of Z. spina-christi L. was 7.5%. PC and HPLC studies revealed that the hydrolysate of the mucilage of Z. spina-christi L. contains, mannose (93%), glucuronic acid (4.4%), rhamnose (1.24%) and galacturonic acid (0.37%). The lipid content of Z. spina-christi L. seeds was studied. Analysis of the fatty acids, by a Gas Liquid Chromatography (GLC) technique, revealed the presence of 13 fatty acids; linoleic acid represents 45% of the total mixture, followed by linolenic acid (20%). Cholesterol and β-sitosterol represent the major constituents of the unsaponifiable fraction (ca. 49%). This lipid fraction showed antimicrobial activity against G+ve Bacillus subtilis , and Streptococcus pyogenes , G−ve Escherichia coli . Fatty acid fraction showed high activity against E. coli, and B. subtilis .

The polysaccharide chains and the crystallinity of beta-glucan in a white sorghum variety, SK(5912) were investigated using chemical and enzymic studies. Mild periodate oxidation and methylation, coupled to descending paper chromatography of products revealed the presence of unresolved non-carbohydrate moiety, 2, 4-and 2, 3-di-O-methyl D-glucose residues (molar ratio; 18:3) and 2, 4, 6-and 2, 3, 6-tri-O-methyl D-glucose residues (molar ratio; 1:14). Paper chromatography of the total acid hydrolysate also revealed a non-carbohydrate spot, identified as protein on the basis of positive Biuret and ninhydrin tests. The O-methyl D-glucose residues suggest two polysaccharide chains designated X and Y. Chain X is formed through linking of beta-D-glucopyranosyl residues by (1-->3) linkages with 85-86% (1-->6) bonds at branch points and constitute about 6-7% of the beta-glucan sample. Chain Y, which is 93-94% of the beta-glucan polysaccharide chains, constitutes beta-D-glucopyranosyl residues in (1-->4) linkages and 4-5% (1-->6) bonds at branch points. Of the 18 branch points on the X-chains in a given beta-glucan sample, about 15 are the Y chains interlinked to the X-chains through their (Y-chains) reducing ends. Both acid and enzyme hydrolyses of the beta-glucan suggest two structural organizations, a crystalline and less crystalline granules, based on two first order kinetics. This was correlated by the progress curves obtained during hydrolysis with two purified isoforms of beta-glucanases from the sorghum malt. The short and highly branched polysaccharide chains, and longer but less branched polysaccharide chains found in this beta-glucan are reminiscent of the structures of amylopectin and amylose, respectively. The K(m)s of 0.30-0.32 and 0.42-0.50 mg beta-glucan/ml for the beta-glucanase isoforms also lay credence to both the crystalline forms and the highly polymerised nature of the beta-glucan in white sorghum.

Aspergillus niger could utilize D-galactose as sole source of carbon. Cell-free extracts of D-galactose-grown mycelia were able to catalyze the oxidation of D-galactose to D-galactonic acid-gamma-lactone (GalA-gamma-lact) in the presence of NAD, followed by the appearance of 2-keto-3-deoxy-D-galactonate (KDGal), pyruvate and glyceraldehyde. From 10mgr;moles only 6.6mgr;moles of GalA-gamma-lact were disappeared after 60 min of reaction indicating the presence of GalA-gamma-lactonase. Identification of GalA-gamma-lact was achieved by ascending paper chromatography. KDGal, pyruvate and glyceraldehyde were also chromatographically identified in the reaction mixture containing D-galactonate which suggests that D-galactonate is degraded into pyruvate and glyceraldehyde via the intermediate formation of KDGal. Such reactions are supposed to be catalyzed by an inducible D-galactonate dehydratase and a constitutive KDGal aldolase. The amount of KDGal, pyruvate and glyceraldehyde were found to be almost equivalent and the equilibrium of the reaction being toward the formation of KDGal. The apparent equilibrium constant (K(eq)) was calculated and found to be 0.5 x 10(-3) M. Results also proved the reversibility of the reaction catalyzed by KDGal aldolase of A. niger. In the light of the findings obtained from the degradation of D-galactose by cell-free extracts of A. niger grown on D-galactose and D-galactonate a nonphosphorolytic pathway was suggested to be operative for the degradation of D-galactose in extracts of A. niger.

188Re is one of the radioisotopes expected to emerge as useful for therapy. Development of new radiopharmaceuticals based on 188Re depends on the radiolabeling methods used, which would give stable complexes having predefined radiochemical properties and in vitro and in vivo stability. This paper has attempted to provide a perspective of 188Re-labeled monoclonal antibodies, their radiolabeling characteristics, methods for quality control of radioimmunoconjugates and in vitro stability for radioimmunotherapy of solid tumors. The direct method of 188Re radiolabeling of antibodies by reductive attachment of 188Re in which free sulfhydryl groups have been generated by reduction of the intramolecular S–S disulfide bonds has been shown to be a promising approach in particular. Moreover, excellent methods have been developed to test the radionuclide, radiochemical purity and stability of 188Re-radioimmunoconjugates using high performance liquid chromatography (HPLC) and paper chromatography.

To reveal the metabolic fate of acetate in neoplasms that may characterize the accumulation patterns of [1-(11)C]acetate in tumors depicted by positron emission tomography. Four tumor cell lines (LS174T, RPMI2650, A2780, and A375) and fibroblasts in growing and resting states were used. In uptake experiments, cells were incubated with[1-(14)C]acetate for 40 min. [(14)C]CO(2) was measured in the tight-air chamber, and the metabolites in cells were identified by thin layer chromatography and paper chromatography. The glucose metabolic rate of each cell line was measured with [2,6-(3)H]2-deoxy-glucose (DG), and the growth activity of each cell line was estimated by measuring the incorporation of [(3)H]methyl thymidine into DNA. Compared with resting fibroblasts, all four tumor cell lines showed higher accumulation of (14)C activity from [1-(14)C]acetate. These tumor-to-normal ratios of [1-(14)C]acetate were larger than those of DG. Tumor cells incorporated (14)C activity into the lipid-soluble fraction, mostly of phosphatidylcholine and neutral lipids, more prominently than did fibroblasts. The lipid-soluble fraction of (14)C accumulation in cells showed a positive correlation with growth activity, whereas the water-soluble and CO(2) fractions did not. These findings suggest that the high tumor-to-normal ratio of [1-(14)C]acetate is mainly due to the enhanced lipid synthesis, which reflects the high growth activity of neoplasms. This in vitro study suggests that [1-(11)C]acetate is appropriate for estimating the growth activity of tumor cells.

Carbohydrates were extracted from hooded seal milk, Crystophora cristata (family Phocidae). Free oligosaccharides were separated by gel filtration and then purified by ion exchange chromatography, gel filtration and preparative thin layer or paper chromatography and their structures determined by 1H-NMR. The hooded seal milk was found to contain inositol and at least nine oligosaccharides, most of which had lacto-N-neotetraose or lacto-N-neohexaose as core units, similar to those in milk of other species of Carnivora such as bears (Ursidae). Their structures were as follows: Gal(beta1-4)Glc (lactose); Fuc(alpha1-2)Gal(beta1-4)Glc (2'-fucosyllactose); Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc (lacto-N-neotetraose); Fuc(alpha1-2)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc (lacto-N-fucopentaose IV); Gal(beta1-4)GlcNAc(beta1-3)[Gal(beta1-4)GlcNAc(beta1-6)]Gal(1-4)Glc (lacto-N-neohexaose); Fuc(alpha1-2)Gal(beta1-4)GlcNAc(beta1-3)[Gal(beta1-4)GlcNAc(beta1-6)]Gal(beta1-4)Glc (monofucosyl lacto-N-neohexaose a); Gal(beta1-4)GlcNAc(beta1-3)[Fuc(alpha1-2)Gal(beta1-4)GlcNAc(beta1-6)]Gal(beta1-4)Glc (monofucosyl lacto-N-neohexaose b); Fuc(alpha1-2)Gal(beta1-4)GlcNAc(beta1-3)[Fuc(alpha1-2)Gal(beta1-4)GlcNAc(beta1-6)]Gal(beta1-4)Glc (difucosyl lacto-N-neohexaose); Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc (para lacto-N-neohexaose); Fuc(alpha1-2)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc (monofucosyl para lacto-N-neohexaose). Milk of the Australian fur seal, Arctophalus pusillus doriferus (family Otariidae) contained inositol but no lactose or free oligosaccharides. These results, therefore, support the hypothesis that the milk of otariids, unlike that of phocids, contains no free reducing saccharides.

The syntheses of four alpha-aminomethyl phosphonates and their complexation studies with (99m)Tc and (186/188)Re are reported. Complexation conditions were standardized to give maximum yields, which ranged from 90-97%. The yields of complexation were estimated by paper chromatography. The (99m)Tc complexes were stable for more than 4 h, while the (186/188)Re complexes were stable for 3-8 days when stored at 4 degrees C. Biodistribution of these complexes in Wistar rats were carried out, and the uptake in bone and other soft tissue are detailed. Bone uptake of the (99m)Tc complexes varied from 40-60% at 30 min postinjection depending on the ligands. The uptake in soft tissue was minimum with all the complexes. A comparison of the biodistribution studies of the (99m)Tc complexes with that of the well-established radiopharmaceutical (99m)Tc-MDP was carried out for the purpose of evaluating the efficacy of the radiopharmaceutical preparation with the complexes of these ligands. The bone uptake of the (186/188)Re complexes varied from 19-28% corresponding to 1.6-3% per g at 3 h postinjection. The residual activity in both (99m)Tc and (186/188)Re complexes showed renal clearance.

A microbial biosensor for sugar determination was prepared by surface modification of a graphite electrode using Gluconobacter oxydans cells. The sensitivity of amperometric detection was enhanced by using hexacyanoferrate(III) as a mediator. The G. oxydans cells contain membrane-bound aldose dehydrogenase, which catalyses the oxidation of wide range of sugars including all sugars present in lignocellulose hydrolysate. The substrate specificity of the biosensor, effect of pH, temperature, working potential, hexacyanoferrate(III) concentration as well as the physiological state of the cells for detection were carefully optimised. The upper value of the linear range of the optimised biosensor was in the range 1.1–2.2 g l −1 for determination of d -glucose, d -galactose, d -xylose, d -mannose and l -arabinose. The biosensor was used for total sugars determination during lignocellulose hydrolysate fermentation. A good correlation between total sugars determined in samples by the biosensor and by quantitative paper chromatography was obtained.

An overview of analytical methods for the measurement of biologically important tea catechins is presented. Liquid chromatography and capillary electrophoresis are the most cited techniques for catechin separation, identification and quantitation. Liquid chromatography with ultraviolet detection is frequently used; however, mass spectrometry, electrochemical, fluorescence and chemiluminescence detection are also utilized in cases where more sensitive or selective detection is needed. Two modes of capillary electrophoresis, capillary zone electrophoresis and micellar electrokinetic capillary chromatography, have been employed for the determination of catechins. Both modes of capillary electrophoresis are based on ultraviolet detection. Additional analytical techniques, such as gas chromatography, thin-layer chromatography, paper chromatography, spectrophotometry, biosensing, chemiluminescence and nuclear magnetic resonance spectroscopy have also been utilized for the determination of catechins and are reviewed herein.

A simple method of preparation of a new therapeutic colloid, samarium(III) phosphate- 153 Sm (SMPC), is reported involving the reaction of carrier-added 153 SmCl 3 with phosphoric acid. Recovery of the colloid was accomplished by dialysis leading to purification and a radiochemical (RC) yield of more than 90%. The RC purity of purified colloid formulated in isotonic phosphate buffer was more than 99% as assessed by paper chromatography. The product retained its RC purity throughout the period of stability study of 7 days. Complete retention of radioactivity instilled in the rabbit knee joint was observed over the study period of 6 days, with radioactivity in the blood being indistinguishable from the natural background activity. Ninety-six percent of colloidal particles were in the size range of 0.3–2 μm. The promising results demonstrated warrant further studies on SMPC for assessing the suitability for therapy.

Evanescent-wave optics in highly turbid media induces peculiar photochemical reactions in polynucleotides. A singularity is observed and appraised experimentally in a quantitative way by irradiation in the UV-C spectral range of dry chromatography paper disks previously impregnated with ribonucleic acid (RNA) solutions, also containing 0.01 to 1.5 M NaCl. Bichromatic transmission photometry of the disks is based on multiple-light-scattering enhanced absorbance measurements. The calculated bleaching cross section (σ 2 ) of the RNA uracil bases situated within the penetration depth of the evanescent wave surrounding the cellulose fibres is over 30 times lower than the cross section (σ 1 ) corresponding to the uracil residues of the polynucleotide chain segments outside the evanescent-wave domain.

The biosynthesis of Acetobacter xylinum ATCC 10821 cellulose has been studied with resting cells and a membrane preparation using 14C-pulse and chase reactions, with d -glucose and UDPGlc, respectively. Cellulose was biosynthesized from UDPGlc, and it was found to be tightly associated with both the cells and the membrane. The cellulose chains could be released from the cells and the membrane preparation by treating at pH 2, 100 °C for 20 min. The cellulose chains that were released from the pulse and pulse-chase reactions were purified and separated from any low molecular weight substances by gel chromatography on Bio-Gel P4. They were then reduced with sodium borohydride and hydrolyzed with 4 M trifluoroacetic acid at 121 °C for 2 h. Labeled products from the acid hydrolyzates were separated by paper chromatography and found to be d -glucose and d -glucitol. The amount of radioactivity in the products was determined by liquid scintillation counting. It was found that the pulsed products from the resting cells gave a ratio of d -[14C]glucitol to d -[14C]glucose of 1:11, and after chasing, the ratio decreased to 1:36. The pulsed products from the membrane gave a ratio of d -[14C]glucitol to d -[14C]glucose of 1:12, and after chasing for 5 min the ratio decreased to 1:43, and after 10 min, the ratio decreased to 1:66. These results show that the labeled d -glucitol obtained from the reducing end of the cellulose chain is chased into the interior of the cellulose chain during synthesis, showing that the cellulose chain is elongated from the reducing end. An insertion mechanism for the synthesis of cellulose from UDPGlc is proposed that involves lipid pyrophosphate glycosyl intermediates and three membrane enzymes: lipid phosphate:UDPGlc phosphotransferase, cellulose synthase, and lipid pyrophosphate phosphohydrolase.

A nonaromatic broadspectrum antibiotic of Streptomyces antibioticus Sr 15. 4 was purified by adsorption on activated charcoal, paper chromatography and preparative TLC. UV, Mass and IR spectra revealed that the antibiotic of the strain Sr 15. 4 is distinctly different of nonaromatics having alcoholic-OR and carbonyl group.