Determination of sugars and sugar alcohols in infant formula by high performance liquid chromatography with evaporative light-scattering detector.

A method was established for the simultaneous determination of five sugars (fructose, glucose, sucrose, lactose, maltose) and five sugar alcohols (erythritol, xylitol, sorbitol, mannitol, maltitol) in infant formula by high performance liquid chromatography-evaporative light scattering detector. After the samples were extracted with acetonitrile-water solution, precipitated by acetic acid, and purified with solid phase extraction cartridge, ALLChrom Rocksil Carbohydrate ES column was adopted for separation, and isocratic elution was conducted at the flow rate of 1.0 mL/min with acetonitrile-0.04 % ammonia solution as the mobile phase. The analytes were detected by an evaporative light-scattering detector, and quantified by external standard method. The linear ranges of the 10 components were 0.04-4.0 g/L with the correlation coefficients greater than 0.999, and the limits of quantification (S/N = 10) of the method were 0.08-0.4 g/100 g. The relative standard deviation of the lactose parallel samples reached 1.29 %, and the recoveries of the other 9 components ranged from 80.4 % to 99.4 % with the relative standard deviation of 2.8 %-7.1 %. The method performs well in sensitivity and separation, which is suitable for the simultaneous quantitative determination of sugars and sugar alcohols in infant formula.

[Sugar and sugar alcohols in infants Ying Yang Bao based on derivatization and gas chromatography-mass spectrometry].

OBJECTIVE: To establish a gas chromatography-mass spectrometry(GC-MS) method for determination of seven sugars and sugar alcohols in infants Ying Yang Bao nutritional supplements. METHODS: The samples were extracted with pure water and diluted with 95% ethanol. After being dried by nitrogen, methoxyamine hydrochloride oxime was dissolved in pyridine and derivatized by MSTFA. The capillary column TG-5 Ms(30 m×0. 25 mm, 0. 25 µm) was used for determination by GC-MS. RESULTS: The limits of detection(LODs)were 1. 0-3. 0 mg/g and the limits of quantification(LOQs)were 3. 3-10. 0 mg/g. The average recoveries of seven kinds of sugar and sugar alcohols were 86. 7%-96. 7%, and the relative standard deviation was less than 5. 1%(n=6). The contents of seven sugars and sugar alcohols in soybean matrix nutritional supplements were determined in the range of 0. 25-13. 70 g/100 g, which was consistent with the nutrition label of the products. CONCLUSION: The method is convenient, mild and fit for batch sample analysis.

Development of Alditol Acetate Derivatives for the Determination of 15N-Enriched Amino Sugars by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry.

Amino sugars can be used as indices to evaluate the role of soil microorganisms in active nitrogen (N) cycling in soil. This paper details the assessment of the suitability of gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) for the analysis of 15N-enriched amino sugars as alditol acetate derivatives prior to application of a novel 15N stable isotope probing (SIP) approach to amino sugars. The efficient derivatization and cleanup of alditol acetate derivatives for GC was achieved using commercially available amino sugars, including glucosamine, mannosamine, galactosamine, and muramic acid, as laboratory standards. A VF-23ms stationary phase was found to produce optimal separations of all four compounds. The structure of the alditol acetate derivatives was confirmed using gas chromatography/mass spectrometry (GC/MS). For GC-C-IRMS determinations, implementation of a two-point normalization confirmed the optimal carrier gas flow rate to be 1.7 mL min-1. Linearity of δ15N value determinations up to δ15Nt of 469 ± 3.1‰ (where δ15Nt is the independently measured δ15N value) was confirmed when 30 nmol N was injected on-column, with the direction of deviation from δ15Nt at low sample amount dependent on the 15N abundance of the analyte. Observed between- and within-run memory effects were significant ( P < 0.007) when a highly enriched standard (469 ± 3.1‰) was run; therefore, analytical run order and variation in 15N enrichment of analytes within the same sample must be considered. The investigated parameters have confirmed the isotopic robustness of alditol acetate derivatives of amino sugars for the GC-C-IRMS analysis of 15N-enriched amino sugars in terms of linearity over an enrichment range (natural abundance to 469 ± 3.1‰) with on-column analyte amount over 30 nmol N.

An isotope dilution ultra high performance liquid chromatography-tandem mass spectrometry method for the simultaneous determination of sugars and humectants in tobacco products.

CDC's Division of Laboratory Sciences developed and validated a new method for the simultaneous detection and measurement of 11 sugars, alditols and humectants in tobacco products. The method uses isotope dilution ultra high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and has demonstrated high sensitivity, selectivity, throughput and accuracy, with recoveries ranging from 90% to 113%, limits of detection ranging from 0.0002 to 0.0045µg/mL and coefficients of variation (CV%) ranging from 1.4 to 14%. Calibration curves for all analytes were linear with linearity R2 values greater than 0.995. Quantification of tobacco components is necessary to characterize tobacco product components and their potential effects on consumer appeal, smoke chemistry and toxicology, and to potentially help distinguish tobacco product categories. The researchers analyzed a variety of tobacco products (e.g., cigarettes, little cigars, cigarillos) using the new method and documented differences in the abundance of selected analytes among product categories. Specifically, differences were detected in levels of selected sugars found in little cigars and cigarettes, which could help address appeal potential and have utility when product category is unknown, unclear, or miscategorized.

Characterization of Sugar and Polyphenolic Diversity in Floral Nectar of Different 'Oblacinska' Sour Cherry Clones.

'Oblacinska' sour cherry, an autochthonous cultivar, is the most planted cultivar in Serbian orchards. Since fruit trees in temperate zone reward insects by producing nectar which 'quality' affects the efficiency of insect pollination, the aim of this study was analyzing of sugars and polyphenolics in floral nectar of 16 'Oblacinska' sour cherry clones with different yielding potential. The contents of sugars and sugar alcohols were analyzed by ion chromatography, while polyphenolic profile was established using liquid chromatography/mass spectrometry technique. Fourteen sugars and six sugar alcohols were detected in nectar samples and the most abundant were fructose, glucose, and sucrose. Eleven polyphenols were quantified using available standards, while another 17 were identified according to their exact masses and characteristic fragmentations. Among quantified polyphenols, rutin, naringenin, and chrysin were the most abundant in nectar. Principal component analysis showed that some polyphenol components (naringin, naringenin, and rutin) together with sugars had high impact of spatial distribution of nectar samples on score plot.

[Determination of alditols in wine by gas chromatography-mass spectrometry after acetate derivatization].

The acetate derivatization of alditols for determining alditol level in wine by gas chromatography (GC)-mass spectrometry (MS) has been developed. The wine sample was mixed with pyridine and centrifuged at 5,000 r/min at the temperature of 4 degrees C for 10 min. After filtration with organic phase membrane, the supernatant was derivatized with acetic anhydride, and then dehydrated with anhydrous sodium sulfate. The GC separation was performed on a DB-5MS capillary column. The alditols were determined by MS in selected ion monitoring (SIM) mode and quantified by external standard method. The calibration curves showed good linearities in the range of 0.019 - 1.25 mg/L except for lactitol (0.039 - 2.50 mg/L) with the correlation coefficients greater than 0.99. The limits of quantification (S/N= 10) of erythritol, xylitol, D-mannitol, sorbitol, galactitol and lactitol were 0.17, 0.29, 0.43, 0.46, 0.47 and 2.88 mg/L respectively. The limits of detection (S/N = 3) were 0.05, 0.08, 0.13, 0.14, 0.14 and 1.38 mg/L respectively. The recoveries of alditols spiked in the wine at two levels of 40 mg/L and 80 mg/L were ranged from 80.15% to 108.75% with the relative standard deviations (RSDs) of 2.16% - 6.97%. The sensitivity, accuracy and precision of the method can meet the technical standard. The method can be applied to the rapid determination of alditols in wine.

Looking beyond sugars: phytochemical profiling and standardization of manna exudates from Sicilian Fraxinus excelsior L.

Different grades of genuine and counterfeit Fraxinus excelsior exudates, marketed as natural sweeteners or mild laxatives, were evaluated for their proximate composition and for saccharidic, organic acids, lipidic and phenolic profile by means of GC-MS and (1)H NMR. Genuine samples contained mannitol (39-48 g/100 g, according to the grade), fructose (9-16 g/100 g), glucose (2-3.7 g/100 g), sorbitol (0,5-0,6 g/100 g), galactose (0.02-0.74 g/100 g), oligosaccharides as mannotriose (13-22 g/100 g) and stachyose (1-11 g/100 g), and traces of myo-inositol, mannose, sucrose. On the contrary, counterfeit samples contained mostly mannitol and sorbitol, with traces of fructose, glucose and mannose. Differences in ash, total polyphenolic content and fatty acid composition allowed a quick identification of counterfeit products, confirmed by a distinct mono-, oligosaccharidic and phenolic pattern. Elenolic acid (63-1628 mg/kg), tyrosol (15-774 mg/kg), homovanillic acid (2,39-52.8 mg/Kg), dopaol (0.8-63 mg/kg), pinoresinol (4.2-18.5 mg/kg) and fraxetin (0.25-11.64 mg/kg), albeit showing a wide concentration range, were the most abundant substances detected in the phenolic fraction of Fraxinus manna, while esculetin, p-hydroxybenzoic acid, 4-hydroxyphenacetic acid, 3,4 hydroxybenzoic acid, hydroxy-pinoresinol, medioresinol and siringaresinol were present in low amounts. The polyphenolic profile may be used as a marker for authentication and should be considered in the evaluation of nutritional and health properties ascribed to Fraxinus manna.

Metabolic profiling during peach fruit development and ripening reveals the metabolic networks that underpin each developmental stage.

Fruit from rosaceous species collectively display a great variety of flavors and textures as well as a generally high content of nutritionally beneficial metabolites. However, relatively little analysis of metabolic networks in rosaceous fruit has been reported. Among rosaceous species, peach (Prunus persica) has stone fruits composed of a juicy mesocarp and lignified endocarp. Here, peach mesocarp metabolic networks were studied across development using metabolomics and analysis of key regulatory enzymes. Principal component analysis of peach metabolic composition revealed clear metabolic shifts from early through late development stages and subsequently during postharvest ripening. Early developmental stages were characterized by a substantial decrease in protein abundance and high levels of bioactive polyphenols and amino acids, which are substrates for the phenylpropanoid and lignin pathways during stone hardening. Sucrose levels showed a large increase during development, reflecting translocation from the leaf, while the importance of galactinol and raffinose is also inferred. Our study further suggests that posttranscriptional mechanisms are key for metabolic regulation at early stages. In contrast to early developmental stages, a decrease in amino acid levels is coupled to an induction of transcripts encoding amino acid and organic acid catabolic enzymes during ripening. These data are consistent with the mobilization of amino acids to support respiration. In addition, sucrose cycling, suggested by the parallel increase of transcripts encoding sucrose degradative and synthetic enzymes, appears to operate during postharvest ripening. When taken together, these data highlight singular metabolic programs for peach development and may allow the identification of key factors related to agronomic traits of this important crop species.

Metabolite profiling of sucrose effect on the metabolism of Melissa officinalis by gas chromatography-mass spectrometry.

The effect of sugar on plant metabolism, which is known to be similar to hormone-like signaling, was metabolomically studied using Melissa officinalis (lemon balm). The metabolite profiles of M. officinalis treated with sucrose were analyzed by gas chromatography-mass spectrometry (GC-MS) and principal component analysis (PCA). A total of 64 metabolites from various chemical classes including alcohols, amines, amino acids, fatty acids, inorganic acids, organic acids, phosphates, and sugars were identified by GC-MS. Three groups treated with different sucrose concentrations were clearly separated by PCA of their metabolite profiles, indicating changes in the levels of many metabolites depending on the sucrose concentration. Metabolite profiling revealed that treatment with a higher sucrose level caused an increase in the levels of metabolites such as sugars, sugar alcohols, and sugar phosphates, which are related to the glycolytic pathway of M. officinalis. Furthermore, proline and succinic acid, which are associated with the proline-linked pentose phosphate pathway, the shikimic acid pathway, and the biosynthesis of phenylpropanoids, also increased with increasing sucrose concentration. Therefore, these metabolic changes induced by sucrose ultimately led to the increased production of flavonoids such as caffeic acid via the biosynthetic pathway of phenylpropanoids. This study demonstrated that the abundance changes in some primary and secondary metabolites were somewhat interlocked with each other in response to sucrose.

A metabolomics study of cultivated potato (Solanum tuberosum) groups Andigena, Phureja, Stenotomum, and tuberosum using gas chromatography-mass spectrometry.

Phytochemical diversity was examined by gas chromatography-mass spectrometry in tubers of genotypes belonging to groups Andigena, Phureja, Stenotomum, and Tuberosum of the potato, Solanum tuberosum. Polar extracts (mainly amino acids, organic acids, sugars, and sugar alcohols) and nonpolar extracts (mainly fatty acids, fatty alcohols, and sterols) were examined. There was a large range in levels of metabolites, including those such as asparagine, fructose, and glucose, that are important to tuber quality, offering considerable scope for selecting germplasm for breeding programmes. There were significant differences in the levels of many metabolites among the groups. The metabolite profiles of genotypes belonging to Phureja and Stenotomum were similar and different from those of Tuberosum and the majority of Andigena genotypes. There was some agreement with the phylogeny of the groups in that Stenotomum is believed to be the ancestor of Phureja and they are both distinct from Tuberosum. Andigena genotypes could be partially distinguished according to geographical origin, Bolivian genotypes being particularly distinct from those from Ecuador. Biosynthetic links between metabolites were explored by performing pairwise correlations of all metabolites. The significance of some expected and unexpected strong correlations between many amino acids (e.g., between isoleucine, lysine, valine, and other amino acids) and between several nonpolar metabolites (e.g., between many fatty acids) is discussed. For polar metabolites, correlation analysis gave essentially similar results irrespective of whether the whole data set, only Andigena genotypes, or only Phureja genotypes were used. In contrast, for the nonpolar metabolites, Andigena only and Phureja only data sets resulted in weaker and stronger correlations, respectively, compared to the whole data set, and may suggest differences in the biochemistry of the two groups, although the interpretation should be viewed with some caution.

El lactitol incrementa el glutation reducido y disminuye el óxido nítrico en ratas Sprague-Dawley / Lactitol increases reduced glutathione and decreases nitric oxide in Sprague Dawley rats

Existe un creciente uso de los alcohol-azúcares como el lactitol en la industria de los alimentos. El estrés oxidativo juega un papel importante en la génesis de patologías digestivas que van desde inflamación hasta cáncer. El propósito de este estudio fue determinar el efecto del lactitol sobre el malondialdehído (MDA), óxido nítrico (NO), glutation reducido (GSH), ácido ascórbico y ácido dehidroascórbico como marcadores del balance oxidación/antioxidación. Para ello se utilizaron 80 ratas macho Sprague-Dawley divididas en cuatro grupos , tres experimentales de 20 animales, a los cuales se les administró por sonda orogástrica, lactitol en dosis de 0,3; 1,0 y 5,0 g/Kg/día durante 12 semanas y un grupo control que recibió solución salina fisiológica por el mismo período de tiempo. El lactitol administrado en dosis de 0,3; 1,0 y 5,0 g/Kg/día produjo un incremento significativo (P<0,05) del GSH (326,5 ± 13,0 µg/ml; 328,5 ± 9,2 µg/ml y 398,2 ± 11,8 µg/ml) al ser comparado con sus respectivos valores basales (285,8 ± 4,0 µg/ml; 280,0 ± 6,2 µg/ml y 279,5 ± 9,1 µg/ml). El lactitol a dosis de 5 g/Kg/día produjo el más alto incremento de la concentración de GSH y al mismo tiempo provocó una disminución significativa del los niveles de NO (33,0 ± 1,2 µM) cuando se comparó con su concentración basal (46,2 ± 2,8 µM). No fueron observados cambios significativos sobre el resto de los marcadores del balance oxidación/antioxidación. Aunque el lactitol es un alcohol-azúcar que no se absorbe a nivel del tracto gastrointestinal, es posible que los productos finales obtenidos luego de su metabolismo por las bacterias intestinales, induzcan efectos sistémicos que pueden afectar el balance oxidación/antioxidación a favor de la antioxidación.

Sugar alcohols such as lactitol are increasingly being used in the food industry. Tissue oxidative stress is an important contributor to the genesis of inflammatory bowel disease and cancer. The purpose of this study was to determine the effect of lactitol on malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide (NO), dehydroascorbic and ascorbic acid as redox markers. Eighty Sprague Dawley rats were divided into four groups; three experimental groups which received lactitol through an oral catheter at doses of 0.3; 1.0; 5 g/kg/day and an experimental group to which saline solution was administered during 12 weeks. Lactitol at doses of 0.3; 1.0; 5 g/kg/day produced a significant increase (P<0.05) on GSH (326.5 ± 13.0 µg/ml; 328.5 ± 9.2 µg/ml y 398.29 ± 11.8 µg/ml respectively) when compared with their respective basal values (285.8 ± 4.0 µg/ml; 280.0 ± 6.2 µg/ml y 279.5 ± 9.1 µg/ml). Lactitol dose of 5g/kg/day produced the highest increase on GSH levels and at the same time elicited a significant decrease on NO levels (33.0 ± 1.2 µM) when compared with basal values (46.2 ± 2.8 µM). No significant changes were observed on the remaining redox markers. Although lactitol is a sugar alcohol that is not absorbed in the small bowel, it is possible that its metabolisms end products, under intestinal bacterial effects, alter the redox balance in favor of antioxidants.

Determination of sugars and alditols in tobacco with high performance anion-exchange chromatography.

Sugars, alditols, and alcohols in tobacco products were quantified utilizing a method of high-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD). The optimized analytical method can be used to classify different tobaccos and control the quality of tobaccos. To substantiate the applicability of the method, the analysis of 27 tobacco samples with satisfactory linearity, repeatability, and accuracy had been demonstrated. Compared with some other analytical methods for tobacco, the HPAEC-PAD method provided a simple and powerful tool for the analysis of not only sugars but also alcohols in tobacco extracts.

Development and application of a rapid diagnostic method for invasive Candidiasis by the detection of D-/L-arabinitol using gas chromatography/mass spectrometry.

A rapid non-culture-based diagnostic method utilizing d-/l-arabinitol (DA/LA) ratios as a chemical marker of invasive candidiasis was developed and explored. The enantiomers-ratios detection was made possible by the use of gas chromatography coupled with mass spectrometry (GC/MS). The mean DA/LA ratios +/- standard deviation (range) in urine (n = 40) and serum (n = 20) were 2.08 +/- 0.78 (0.57 to 3.55) and 1.79 +/- 0.75 (0.74 to 3.54), respectively, from patients without evidence of fungal infection or colonization; in patients (n = 7) with culture-proven invasive candida infections, the figures were 9.91 +/- 3.04 (7.24 to 16.27) and 13.58 +/- 7.31 (5.57 to 25.88) in urine and serum, respectively. The differences in DA/LA ratios between the candidemic patients and the non-candidemic patients were statistically significant (p < 0.01) in both serum and urine samples. The DA/LA ratios were not significantly affected in patients with oral or vaginal candidiasis and candiduria.

Determination of amino acids in cell culture and fermentation broth media using anion-exchange chromatography with integrated pulsed amperometric detection.

Cell culture and fermentation broth media are used in the manufacture of biotherapeutics and many other biological materials. Characterizing the amino acid composition in cell culture and fermentation broth media is important because deficiencies in these nutrients can reduce desired yields or alter final product quality. Anion-exchange (AE) chromatography using sodium hydroxide (NaOH) and sodium acetate gradients, coupled with integrated pulsed amperometric detection (IPAD), determines amino acids without sample derivatization. AE-IPAD also detects carbohydrates, glycols, and sugar alcohols. The presence of these compounds, often at high concentrations in cell culture and fermentation broth media, can complicate amino acid determinations. To determine whether these samples can be analyzed without sample preparation, we studied the effects of altering and extending the initial NaOH eluent concentration on the retention of 42 different carbohydrates and related compounds, 30 amino acids and related compounds, and 3 additional compounds. We found that carbohydrate retention is impacted in a manner different from that of amino acid retention by a change in [NaOH]. We used this selectivity difference to design amino acid determinations of diluted cell culture and fermentation broth media, including Bacto yeast extract-peptone-dextrose (yeast culture medium) broth, Luria-Bertani (bacterial culture medium) broth, and minimal essential medium and serum-free protein-free hybridoma medium (mammalian cell culture media). These media were selected as representatives for both prokaryotic and eukaryotic culture systems capable of challenging the analytical technique presented in this paper. Glucose up to 10mM (0.2%, w/w) did not interfere with the chromatography, or decrease recovery greater than 20%, for the common amino acids arginine, lysine, alanine, threonine, glycine, valine, serine, proline, isoleucine, leucine, methionine, histidine, phenylalanine, glutamate, aspartate, cystine, and tyrosine.

Simultaneous identification and quantification of the sugar, sugar alcohol, and carboxylic acid contents of sour cherry, apple, and ber fruits, as their trimethylsilyl derivatives, by gas chromatography-mass spectrometry.

Our gas chromatography-mass spectrometry method--developed for the simultaneous quantitation of mono-, di-, and trisaccharides, sugar alcohols, caboxylic and amino acids, measured as their trimethylsilyl-(oxime) ether/ester derivatives, from one solution by a single injection, prepared in the presence of the fruit matrix--has been extended/utilized for special purposes. The compositions of (i) freshly harvested and stored sour cherries (Prunus cerasus), (ii) apples obtained from organic and integrated productions (Malus domestica), and (iii) green and ripe bers (Zizyphus mauritiana L.) were compared. On the basis of earlier, basic researches (derivatization, quantitation, and fragmentation studies of authentic compounds), we demonstrate the reproducible quantitation of the main and minor constituents in a wide concentration range (approximately 1 x 10(-)(3) to >/=40%, in total up to < or =98%, calculated on dry matter basis of the fruit matrices). Reproducibility of quantitations, calculated on the basis of their total ion current values, provided an average reproducibility of 3.3 (sour cherries), 6.2 (apple), and 4.3 (ber) RSD %, respectively.

A headspace solid-phase microextraction method for the determination of some secondary compounds of Brazilian sugar cane spirits by gas chromatography.

A headspace solid-phase microextraction (SPME) method was developed for the determination of secondary compounds from Brazilian sugar cane spirits, or cachaça, by GC-FID. An SPME holder with an 85 microm polyacrylate coating was utilized. The novel method is compared with an optimized method: liquid-liquid extraction (LLE). Both methods showed good linearity, but the repeatability for analyses done with the SPME technique (%RSD = 1.8-3.9) was better than for those done with LLE (%RSD = 10.3-11.7). The concentrations of the analytes obtained in the analysis of 12 cachaça samples with the SPME technique were higher than those obtained with LLE. In the SPME method the extraction wastes are smaller. Cachaça samples were qualitatively analyzed for GC-MS.

Simultaneous determination by capillary gas chromatography of organic acids, sugars, and sugar alcohols in plant tissue extracts as their trimethylsilyl derivatives.

A capillary gas chromatographic (GC) method for the simultaneous determination of organic acids, sugars, and sugar alcohols extracted from plant tissues is described. Plant leaves were extracted in 5% (w/v) perchloric acid and neutralized extracts were purified using C18 cartridges. Organic acids, sugars, and sugar alcohols in purified extracts were converted to their trimethylsilyl (TMS)/TMS-oxime derivatives prior to separation and detection by capillary GC with flame ionization detection (FID). Derivatization procedures were investigated in detail and the compounds of interest were readily converted to their TMS/TMS-oxime derivatives using hexamethyldisiazane reagent in acetonitrile solvent (1:6 v/v) at 100 degreesC for 60 min. The derivatives were sufficiently volatile and stable. The FID response to derivatized compounds was generally linear in the concentration range 30-300 microg ml-1, with detection limits in the order of 3-76 ng. The proposed method was demonstrated for the determination of organic acids, sugars, and sugar alcohols in leaf extracts of two native Australian plants.

Studies on high salt tolerance of transgenic tobacco.

With E. coli mtlD gene (encoding mannitol 1-phosphate dehydrogenase) and gutD gene (encoding glucitol 6-phosphate dehydrogenase) cloned, plant expression vector pBIGM had been obtained by inserting mtlD and gutD genes into binary vector pBin438. Tobacco was transformed with A. tumefaciens LBA4404 containing pBIGM. Results of molecular hybridization of transformed plants indicated that mtlD and gutD genes had integrated into the genomic DNA of tobacco plants. Experiments of salt tolerance and analysis of sugar alcohols showed that the accumulation of different sugar alcohols in transgenic tobacco plants had increased salt tolerance of tobacco.

Capillary gas chromatography combined with ion trap detection for quantitative profiling of polyols in cerebrospinal fluid and plasma.

Polyol species in cerebrospinal fluid and plasma--ribitol, arabitol, xylitol, 1,5-anhydrosorbitol, myo-inositol, mannitol, sorbitol, and galactitol--simultaneously were quantitated by a capillary gas chromatography/ion trap (mass spectrometric) detection method. The details of the methodology are discussed and the results of analysis of polyols in healthy human subjects are reported. Microliter volumes of cerebrospinal fluid or plasma were mixed with internal standard (deuterium labeled myo-inositol), deproteinized, and evaporated to dryness. Polyols were acetylated in the presence of pyridine catalyst and washed with sodium bicarbonate solution and the acetate derivatives were recovered. Standard curve solutions were similarly treated. The polyol components were resolved on a capillary column bonded with 50% phenyl-50% methyl polysiloxane. Chemical ionization mass spectra for the acetate derivatives of polyols were generated in an ion trap using acetonitrile as reagent gas. Each polyol yielded a fragment ion in 100% abundance arising probably from the loss of one acetate moiety from the protonated molecule. These ions were monitored. The relative standard deviation (within-day) for quantitation of polyols was not greater than 8% for cerebrospinal fluid and 15% for plasma matrix. A polyol profile in cerebrospinal fluid and plasma was determined in healthy human subjects and a cerebrospinal fluid/plasma concentration ratio larger than 1.0 was found for all polyol species except 1,5-anhydrosorbitol and xylitol. This assay technique will be used to study the role of polyols in central nervous system diseases.

The carbohydrate structure of the asparagine-linked oligosaccharides of rat plasma thiostatin.

The complete carbohydrate structure of the asparagine-linked oligosaccharides of rat plasma thiostatin was elucidated through chemical and enzymatic methods including gas chromatography-mass spectrometry (GC-MS) and lectin affinity chromatography. Pronase digestion of thiostatin yielded a major glycopeptide fraction with asparagine the most abundant amino acid present. Based on one mole of aspartic acid, the following molar ratios obtained for the four major amino acids: aspartic acid (1.0), threonine (0.53), glycine (0.48) and serine (0.30). Neutral sugar analysis yielded a 3:2 molar ratio for mannose to galactose based on an assigned value to mannose of 3. On this basis, the fraction also contained 3 residues of sialic acid and, on average, 0 to 1 residue of fucose. GC-MS of partially methylated alditol acetates from the glycopeptide fraction identified the presence of biantennary and triantennary structure. Analyses of the neutral sugar and amino-acid composition, together with methylation data, support a biantennary N-linked structure for this major glycopeptide fraction and a triantennary N-linked structure as a lesser component. Sequencing of the desialyated 14C-labelled glycopeptide fraction by sequential exoglycosidase digestion and lectin affinity chromatography uncovered the following saccharide order: terminal galactose, N-acetylglucosamine and pentasaccharide inner core. This sequence is consistent with the N-linked glycan structures demonstrated by methylation and compositional analyses.