Tapping the Full Potential of Infrared Spectroscopy for the Analysis of Technical Lignins.

We present an approach to overcome the challenges associated with the increasing demand of high-throughput characterization of technical lignins, a key resource in emerging bioeconomies. Our approach offers a resort from the lack of direct, simple, and low-cost analytical techniques for lignin characterization by employing multivariate calibration models based on infrared (IR) spectroscopy to predict structural properties of lignins (i. e., functionality, molar mass). By leveraging a comprehensive database of over 500 well-characterized technical lignin samples - a factor of 10 larger than previously used sets - our chemometric models achieved high levels of quality and statistical confidence for the determination of different functional group contents (RMSEPs of 4-16 %). However, the statistical moments of the molar mass distribution are still best determined by size-exclusion chromatography. Analyses of over 500 technical lignins offered also a great opportunity to provide information on the general variability in kraft lignins and lignosulfonates (from different origins). Overall, the effected savings in analysis time (>7 h), resources, and required sample mass combined with non-destructiveness of the measurement satisfy key demands for efficient high-throughput lignin analyses. Finally, we discuss the advantages, disadvantages, and limitations of our approach, along with critical insights into the associated chemical-analytical and spectroscopic challenges.

Profiling of historical rag papers by their non-cellulosic polysaccharide composition.

Hemicellulose and pectin are noteworthy components of historical European rag papers, and have not been studied in detail so far. Rag papers were made from used textiles, and fiber-based utilities, such as ropes and bags. These had been prepared until the mid-19th century from plant-based fibers. Their polysaccharide composition could relate to their condition and history. This information can be expected to hold importance for the preservation and conservation of historical objects. We investigated a collection of rag papers of different age for their composition of non-cellulosic polysaccharides, and compared the findings with modern rag papers and wood pulps. Furthermore, a non-destructive determination of the hemicellulose and pectin content by near-infrared spectroscopy was developed. Historical rag papers had a lower hemicellulose/pectin content than pulps; the fractions of rhamnose, galactose, and arabinose were higher, while xylose was lower. In modern rag papers, xylose tended to be at the higher end of the range, which suggests a degradation of hemicelluloses/pectin over time or a change in raw materials and manufacturing. Rag papers also showed higher crystallinity than wood pulp papers. These findings provide insights into rag paper characteristics and offer potential classification methods.

Cellulose fibers and ellagitannin-rich extractives from rambutan (Nephelium Lappaceum L.) peel by an eco-friendly approach.

This study assesses the viability of an accelerated solvent extraction technique employing environmentally friendly solvents to extract ellagitannins while producing cellulose-rich fibers from rambutan peel. Two sequential extraction protocols were investigated: 1) water followed by acetone/water (4:1, v:v), and 2) acetone followed by acetone/water (4:1, v:v), both performed at 50 °C. The first protocol had a higher extraction yield of 51 %, and the obtained extractives featured a higher total phenolic (531.4 ± 22.0 mg-GAE/g) and flavonoid (487.3 ± 16.9 mg-QE/g) than the second protocol (495.4 ± 32.8 mg-GAE/g and 310.6 ± 31.4 mg-QE/g, respectively). The remaining extractive-free fibers were processed by bleaching using either 2 wt% sodium hydroxide with 3 wt% hydrogen peroxide or 4-5 wt% peracetic acid. Considering bleaching efficiency, yield, and process sustainability, the single bleaching treatment with 5 wt% of peracetic acid was selected as the most promising approach to yield cellulose-rich fibers. The samples were analyzed by methanolysis to determine the amount and type of poly- and oligosaccharides and studied by 13C solid-state nuclear magnetic resonance spectroscopy and thermal gravimetric analysis. The products obtained from the peels demonstrate significant potential for use in various sectors, including food, nutraceuticals, cosmetics, and paper production.

Antimicrobial, antioxidant, and sun protection potential of the isolated compounds from Spermacoce princeae (K. Schum).

BACKGROUND: Spermacoce princeae (K. Schum) has been used in the treatment of bacterial skin infections in Uganda. Pharmacological studies revealed that extracts of S. princeae exhibited antibacterial, antioxidant, and sun protection potential. This study aimed at isolating and identifying pure compounds from the extracts based on comprehensive analytical characterization by multiple analytical techniques. METHODS: The plant samples were extracted by sequential maceration using n-hexane, ethyl acetate, methanol, and distilled water. The compounds were isolated using a combination of chromatographic techniques and their structures were elucidated by multiple spectroscopic techniques. The antibacterial and antifungal activity determination of the isolated compounds was carried out using an agar well diffusion and potato dextrose assay against Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Candida albicans, and Aspergillus flavus while the antioxidant activity was screened with the 2,2-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay. The sun protection factor was determined using a Shimadzu Ultra Violet-visible (UV-VIS) double beam spectrophotometer between 290 to 320 nm. RESULTS: Eleven compounds; quercetin (1), kaempferol-3-O-rutinoside (2), rutin (3, 12), myo-inositol (4), asperulosidic acid (5), hexadecanoic acid (6), ß-sitosterol (7), stigmasterol (8), campesterol (9), ursolic acid (10), and ß-sitosterol glucoside (11) were identified in the S. princeae extracts. Compound 2 had good antifungal activity against C. albicans (zone of inhibition, 23.0 ± 0.1 mm). Compound 10 showed antibacterial and antifungal activity against S. aureus, P. aeruginosa, C. albicans, and A. flavus. Compound 2 had a good percentage radical scavenging effect (IC50 = 64.81 µg/ml) and a good sun protection factor (SPF = 26.83). CONCLUSION: This study reports the first-time isolation and identification of compounds 1 to 11 from S. princeae, which contribute to its antimicrobial, antioxidant, and sun protection potential.

Acetylation of cellulose - Another pathway of natural cellulose aging during library storage of books and papers.

Gaseous acetic acid is formed under conditions of storage of historic paper objects. Its presence not only promotes hydrolytic cleavage of cellulose, but also causes acetylation of the cellulosic material to very small degree. The acetylation reaction proceeds under ambient conditions and without catalyst. Different analytical methods were used to prove the presence of organic acetates on cellulosic paper matrices. DESI-MS in combination with 2H-isotopic labeling showed the presence of sugar fragments with different acetylation patterns. A method based on Zemplen saponification was applied and worked also in the presence of a large excess of acetic acid and/or inorganic acetates. The acetylation effect was quantified for model papers and original, naturally aged paper samples. While cellulose acetylation was clearly proven to be another general pathway of paper aging, further studies of this acetylation phenomenon are needed with regard to conservational aspects and suitable paper storage conditions.

Facile Preparation of Mechanically Robust and Functional Silica/Cellulose Nanofiber Gels Reinforced with Soluble Polysaccharides.

Nanoporous silica gels feature extremely large specific surface areas and high porosities and are ideal candidates for adsorption-related processes, although they are commonly rather fragile. To overcome this obstacle, we developed a novel, completely solvent-free process to prepare mechanically robust CNF-reinforced silica nanocomposites via the incorporation of methylcellulose and starch. Significantly, the addition of starch was very promising and substantially increased the compressive strength while preserving the specific surface area of the gels. Moreover, different silanes were added to the sol/gel process to introduce in situ functionality to the CNF/silica hydrogels. Thereby, CNF/silica hydrogels bearing carboxyl groups and thiol groups were produced and tested as adsorber materials for heavy metals and dyes. The developed solvent-free sol/gel process yielded shapable 3D CNF/silica hydrogels with high mechanical strength; moreover, the introduction of chemical functionalities further widens the application scope of such materials.

Antioxidant properties and qualitative analysis of phenolic constituents in Ephedra spp. by HPTLC together with injection port derivatization GC-MS.

Ephedra herb extracts are being extensively investigated in terms of their antioxidative, antimicrobial and antiproliferative properties, with phenolic components being the general carriers of these bioactivities. Here we describe a comprehensive set of analytical methods employed to determine and characterize both the antioxidative activity and the qualitative profile of phenolic acids and flavonoids present in several Ephedra species of different geographical origin. Spectrophotometric methods were used to determine the total phenolic content, total flavonoid content and antioxidative activity. Multi-development HPTLC enabled chemical fingerprinting which can be used for species differentiation. Individual spots of the thin-layer chromatogram were subjected to GC-MS with injection port derivatization for identification, which was based on both the detected mass spectra and recorded retention indices. The results were compared and complemented with GC-MS using offline derivatization.

Sulfuric Acid-Catalyzed Dehydratization of Carbohydrates for the Production of Adhesive Precursors.

Carbohydrates and hexose-derived 5-hydroxymethylfurfural (5-HMF) are platform chemicals for the synthesis of sustainable binders. New, greener approaches aim at the development of production systems, which minimize process steps and avoid organic solvents or other auxiliaries that could interfere with subsequent resin synthesis. In our work, carbohydrate solutions rich in 5-hydroxymethylfurfural (5-HMF) were produced using a continuous-flow microreactor and diluted H2SO4 as the catalyst. After optimization of the process conditions (temperature, reaction time, catalyst content), a 5-HMF yield of 49% was obtained at a low reaction time of 0.6 min and a catalyst concentration of 1% at 175 °C and 17 bar pressure. Extensive rehydration of the product was avoided by efficient immediate cooling of the reaction solution. The stability of the reaction system was improved by increasing the inner diameter of the capillary in the flow reactor to 2 mm. Advantageously, the obtained reaction mixtures are used directly as precursors in the development of sustainable binder systems, without the need of additional purification, filtration, or extraction steps.

Self-organising maps for the exploration and classification of thin-layer chromatograms.

Thin-layer chromatography (TLC) allows the swift analysis of larger sample sets in almost any laboratory. The obtained chromatograms are patterns of coloured zones that are conveniently evaluated and classified by visual inspection. This manual approach reaches its limit when several dozens or a few hundred samples need to be evaluated. Methods to classify TLCs automatically and objectively have been explored but without a definitive conclusion; established methods, such as principal component analysis, suffer from the variability of the data, while contemporary omics methods were constructed for the analysis of large numbers of highly resolved analyses. Self-organizing maps (SOMs) are an algorithm for unsupervised learning that reduces higher dimensional datasets to a two-dimensional map, locating similar samples close to each other. It tolerates small variations between samples of the same type. We investigated the capability of SOMs for the evaluation of TLCs with two sample sets. With the first one (495 analyses of essential oils), it was confirmed that SOMs arrange the same type of sample in a common region. The obtained multi-class maps were used to classify a test set and to explore the causes for the few misclassifications (<3%). With the second test set (50 extracts of experimental wheats), the effects of a greater variability within substance classes was explored. With SOMs, it was possible to single out the exceptional samples that warranted a more detailed investigation. In addition, the SOM quality control index method was tested. It proved to be considerably stricter than the classification with a SOM of all samples. When this method was unable to classify a sample correctly, it would flag the sample for inspection, as it gave either multiple assignments or none at all. The combination of SOMs and TLC - two accessible analytical tools - can be most useful for the unsupervised classification of samples by TLC, and to identify samples that stand out from a set and are therefore worth the investment into additional analyses with more complex or expensive methods.

Oxidation with a "Stopover" - Stable Zwitterions as Intermediates in the Oxidation of α-Tocopherol (Vitamin E) Model Compounds to their Corresponding ortho-Quinone Methides.

As a prominent member of the vitamin E group, α-tocopherol is an important lipophilic antioxidant. It has a special oxidation chemistry that involves phenoxyl radicals, quinones and quinone methides. During the oxidation to the ortho-quinone methide, an intermediary zwitterion is formed. This aromatic intermediate turns into the quinone methide by simply rotating the initially oxidized, exocyclic methyl group into the molecule's plane. This initial zwitterionic intermediate and the quinone methide are not resonance structures but individual species, whose distinct electronic structures are separated by a mere 90° bond rotation. In this work, we hindered this crucial rotation, by substituting the affected methyl group with alkyl or phenyl groups. The alkyl groups slowed down the conversion to the quinone methide by 18-times, while the phenyl substituents, which additionally stabilize the zwitterion electronically, completely halted the conversion to the quinone methide at -78 °C, allowing for the first time the direct observation of a tocopherol-derived zwitterion. Employing a 13 C-labeled model, the individual steps of the oxidation sequence could be observed directly by NMR, and the activation energy for the rotation could be estimated to be approximately 2.8 kcal/mol. Reaction rates were solvent dependent, with polar solvents exerting a stabilizing effect on the zwitterion. The observed effects confirmed the central relevance of the rotation step in the change from the aromatic to the quinoid state and allowed a more detailed examination of the oxidation behavior of tocopherol. The concept that a simple bond rotation can be used to switch between an aromatic and an anti-aromatic structure could find its use in molecular switches or molecular engines, driven by the specific absorption of external energy.

GC-MS Based Identification of the Volatile Components of Six Astragalus Species from Uzbekistan and Their Biological Activity.

The compositions of volatile components in the aerial parts of six Astragalus species, namely A. campylotrichus (Aca), A. chiwensis (Ach), A. lehmannianus (Ale), A. macronyx (Ama), A. mucidus (Amu) and A. sieversianus (Asi), were investigated using gas chromatograph-mass spectrometry (GC-MS) analysis. Ninety-seven metabolites were identified, accounting for 73.28, 87.03, 74.38, 87.93, 85.83, and 91.39% of Aca, Ach, Ale, Ama, Amu and Asi whole oils, respectively. Sylvestrene was the most predominant component in Asi, Amu and Ama, with highest concentration in Asi (64.64%). In addition, (E)-2-hexenal was present in a high percentage in both Ale and Ach (9.97 and 10.1%, respectively). GC-MS based metabolites were subjected to principal component analysis (PCA) and hierarchal cluster analysis (HCA) to explore the correlations between the six species. The PCA score plot displayed clear differentiation of all Astragalus species and a high correlation between the Amu and Ama species. The antioxidant activity was evaluated in vitro using various assays, phosphomolybdenum (PM), 2,2 diphenyl-1-picryl-hydrazyl-hydrate (DPPH), 2,2-azino bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), cupric reducing antioxidant capacity (CUPRAC), ferric reducing power (FRAP) and ferrous ion chelation (FIC) assays. In addition, the potential for the volatile samples to inhibit both acetyl/butyrylcholinesterases (AChE, BChE), α- amylase, α-glucosidase and tyrosinase was assessed. Most of the species showed considerable antioxidant potential in the performed assays. In the DPPH assay, Ama exhibited the maximum activity (24.12 ± 2.24 mg TE/g sample), and the volatiles from Amu exhibited the highest activity (91.54 mgTE/g oil) in the ABTS radical scavenging assay. The effect was more evident in both CUPRAC and FRAP assays, where both Ale and Ama showed the strongest activity in comparison with the other tested species (84.06, 80.28 mgTE/g oil for CUPRAC and 49.47, 49.02 mgTE/g oil for FRAP, respectively). Asi demonstrated the strongest AChE (4.55 mg GALAE/g oil) and BChE (3.61 mg GALAE/g oil) inhibitory effect. Furthermore, the best tyrosinase inhibitory potential was observed for Ale (138.42 mg KAE/g). Accordingly, Astragalus species can be utilized as promising natural sources for many medicinally important components that could be tested as drug candidates for treating illnesses such as Alzheimer's disease, diabetes mellitus and oxidative stress-related diseases.

The Lipoxygenase Lox1 Is Involved in Light- and Injury-Response, Conidiation, and Volatile Organic Compound Biosynthesis in the Mycoparasitic Fungus Trichoderma atroviride.

The necrotrophic mycoparasite Trichoderma atroviride is a biological pest control agent frequently applied in agriculture for the protection of plants against fungal phytopathogens. One of the main secondary metabolites produced by this fungus is 6-pentyl-α-pyrone (6-PP). 6-PP is an organic compound with antifungal and plant growth-promoting activities, whose biosynthesis was previously proposed to involve a lipoxygenase (Lox). In this study, we investigated the role of the single lipoxygenase-encoding gene lox1 encoded in the T. atroviride genome by targeted gene deletion. We found that light inhibits 6-PP biosynthesis but lox1 is dispensable for 6-PP production as well as for the ability of T. atroviride to parasitize and antagonize host fungi. However, we found Lox1 to be involved in T. atroviride conidiation in darkness, in injury-response, in the production of several metabolites, including oxylipins and volatile organic compounds, as well as in the induction of systemic resistance against the plant-pathogenic fungus Botrytis cinerea in Arabidopsis thaliana plants. Our findings give novel insights into the roles of a fungal Ile-group lipoxygenase and expand the understanding of a light-dependent role of these enzymes.

Unbreakable and customizable dipping chambers for TLC and HPTLC manufactured by fused deposition modelling.

Postchromatographic derivatization chambers for Thin Layer Chromatography (TLC) and its high-performance counterpart (HPTLC) are made of glass, which renders them fragile and - given their frequent use - eventually rather expensive. As a consequence, in daily lab routine, staining reagents are often kept in jars or beakers as cheap and available but unsatisfying equipment. This work describes the design and the manufacturing of shock-resistant and affordable dipping chambers from polypropylene (PP) by fused deposition modelling ("3D printing"). The design is implemented as a customizable script. Three chamber designs are presented: a chamber for typical HPTLC plates (20 × 10 cm) with a convenient closing/storing system which avoids awkward pouring and removal of staining reagents. Second, for synthesis purposes, a chamber for 10 × 10 cm plates with a spout to facilitate exchanging the staining reagent, as well as a very small chamber stabilized by a pedestal for reaction control on 2.5 × 7.5 cm plates, which limits the loss of costly staining reagent. Even though the chambers adsorbed some degraded staining reagent over time, no cross-contamination was observed. All customizable scripts (as CAD models) are made available through online repositories.

The role of PKAc1 in gene regulation and trichodimerol production in Trichoderma reesei.

BACKGROUND: Trichoderma reesei represents a model system for investigation of plant cell wall degradation and its connection to light response. The cyclic adenosine monophosphate pathway (cAMP pathway) plays an important role in both physiological outputs, being crucial for regulation of photoreceptor function as well as for cellulase regulation on different carbon sources. Phosphorylation of photoreceptors and of the carbon catabolite repressor CRE1 was shown in ascomycetes, indicating a relevance of protein kinase A in regulation of the target genes of these transcription factors as well as an impact on regulation of induction specific genes. Moreover, the cAMP pathway impacts growth and development. RESULTS: Here, we investigated gene regulation by the catalytic subunit of protein kinase A (PKAc1) upon growth on cellulose. We found distinct gene sets for regulation upon growth in light and darkness with an overlap of only 13 genes. PKAc1 regulates metabolic genes as well as transport and defense functions. The overlap of gene regulation by PKAc1 with the genes representing the cAMP dependent regulatory output of the photoreceptor ENV1 indicates an involvement of PKA in this pathway, which counteracts its effects by contrasting regulation. Moreover, we found considerable overlap with the gene sets regulated under cellulase inducing conditions and by the carbon catabolite repressor CRE1. Our analysis also showed that PKAc1 regulates the genes of the SOR cluster associated with the biosynthesis of sorbicillinoids. The homologue of gin4, encoding a CAMK type kinase, which is regulated by PKAc1, CRE1 and YPR2 showed a moderate impact on trichodimerol production. We isolated trichodimerol as representative sorbicillin compound and established a method for its quantification in large sample sets using high performance thin layer chromatography (HPTLC), which can be broadly applied for secondary metabolite screening of mutants or different growth conditions. Due to the high expression levels of the SOR cluster under conditions of sexual development we crosschecked the relevance of PKAc1 under these conditions. We could show that PKAc1 impacts biosynthesis of trichodimerol in axenic growth and upon mating. CONCLUSIONS: We conclude that PKAc1 is involved in light dependent regulation of plant cell wall degradation, including carbon catabolite repression as well as secondary metabolism and development in T. reesei.

Structural elucidation of fucoidan from Cladosiphon okamuranus (Okinawa mozuku).

Fucoidan is a sulphated polysaccharide, made up mainly of l-fucose, which is found in brown seaweeds. Its chemical structure is diverse and depends on maturity, species and geographical location. The objective of this study was to elucidate the chemical structure of fucoidan from Cladosiphon okamuranus harvested in Japan. The fucoidan was subject to purification prior to monosaccharide profiling, sulphate content determination, and linkage analysis. Our results showed that Japanese Cladosiphon okamuranus fucoidan contained 70.13 ±â€¯0.22 wt% fucose and 15.16 ±â€¯1.17 wt% sulphate. Other minor monosaccharides found were d-xylose, d-galactose, d-mannose, d-glucose, d-arabinose, d-rhamnose and d-glucuronic acid. Linkage analysis revealed that fucopyranoside units along the backbone are linked, through α-1,3-glycosidic bonds, with fucose branching at C-2, and one sulphate group at C-4 per every three fucose units, i.e. the structure of fucoidan from Japanese Cladosiphon okamuranus is [→3)-α-fuc(1→]0.52[→3)-α-fuc-4-OSO3-(1→]0.33[→2)-α-fuc]0.14.

Flavone glucosides from Artemisia juncea.

A new flavone glucoside, 4',5-dihydroxy-3',5',6-trimethoxyflavone-7-O-ß-D-glucoside was obtained from aerial parts of Artemisia juncea, together with the known flavone eupatilin (5,7-dihydroxy-3',4',6-trimethoxyflavone). The compounds were comprehensively analytically characterized by IR, UV, NMR and HR-MS, and their chemical structures ascertained. The EtOAc fraction of A. juncea showed the strongest DPPH radical scavenging ability as well as reducing power (in CUPRAC and FRAP assays) and phosphomolybdenum activity. This fraction also exhibited the strongest inhibitory effects on tyrosinase. Additionally, the best antidiabetic effects were observed for eupatilin and the CHCl3 fraction.

Profiling and quantification of grain anthocyanins in purple pericarp × blue aleurone wheat crosses by high-performance thin-layer chromatography and densitometry.

BACKGROUND: Anthocyanins are abundant secondary metabolites responsible for most blue to blue-black, and red to purple colors of various plant organs. In wheat grains, anthocyanins are accumulated in the pericarp and/or aleurone layer. Anthocyanin pigmented wheat grains can be processed into functional foods with potential health benefits due to the antioxidant properties of the anthocyanins. The grain anthocyanin content can be increased by pyramidizing the different genes responsible for the accumulation of anthocyanins in the different grain layers. Our objective was to develop a high-performance thin-layer chromatography (HPTLC) method that allows the determination of both the anthocyanin profile and the total pigment concentration. Thereby, selection of breeding lines with significantly higher grain anthocyanin content from purple pericarp × blue aleurone wheat crosses should become more efficient than selection based on only visual scoring of grain color and the unspecific determination of anthocyanin concentration by UV/Vis spectroscopy. RESULTS: A wide variability in the grain anthocyanin content was observed in breeding lines and check varieties. The highest concentration of anthocyanins was observed in deep purple (i.e. combination of the purple pericarp and blue aleurone genetics) grained breeding lines, followed by blue aleurone and purple pericarp genotypes. Determination of the total anthocyanin content was included into the chromatographic analysis, rendering an additional photometric analysis unnecessary. Ten target zones were identified in anthocyanin pigmented wheat grains; four of these zones were typically for blue aleurone types, five for purple pericarp types, and one (i.e. kuromanin glucoside) was characteristic for both. Chemometrics applied to the anthocyanin profile recorded by scanning densitometry revealed that peak heights and peak areas are highly correlated and that seven out of the ten target zones were responsible for about 90% of the total variation in the germplasm. Multivariate analysis of these seven target zones allowed not only a separation of the genetic material into purple, blue and deep purple grained genotypes, but also the identification of genotypes with a specific anthocyanin pattern. Thereby, the original classification by visual scoring was overruled in about one-third of the breeding lines. CONCLUSIONS: The presented HPTLC method with à côté calibration allowed the profiling of the pigments and quantification of wheat grain anthocyanin content in a single analysis, replacing UV/Vis spectroscopy with subsequent HPLC analysis. Moreover, no sample preparation apart from extraction and filtration is required, and more than 15 samples can be evaluated in one analysis run, corresponding to several dozens of samples per day. Hence, the method fulfills the requirements for screening methods in early generations of a plant breeding program such as high-throughput, small sample size, high repeatability, fast determination, and reasonable costs per sample. Combined with multivariate statistical analysis, the anthocyanin pattern allowed the validation of the genetic background in the offspring of purple × blue wheat crosses and, therefore, the efficient selection of genotypes exhibiting both the cyanidin and delphinidin aglycon.

Composition of essential oils from four Apiaceae and Asteraceae species growing in Uzbekistan.

The chemical composition of essential oils isolated from the aerial parts of Heracleum lehmannianum, Prangos pabularia, Pseudohandelia umbellifera and Pulicaria salviifolia, all of them growing in Uzbekistan, were determined by GC-MS analysis. The main components of the oil from H. lehmannianum were α-phellandrene (10.5%), 1-butanol (9.0%), δ-cadinene (6.2%), α-cadinol (5.7%), τ-muurolol (3.1%), 4-terpineol (2.4%) and α-muurolene (2.6%), while cis-allo-ocimene (17.6%), δ-3-carene (14.2%), limonene (7.6%), 2,4,6-trimethylbenzaldehyde (6.8%), α-terpinolene (6.1%), ß-ocimene (4.3%), α-ocimene (4.2%), α-phellandrene (4.2%) were the major oil components in P. pabularia, and borneol (4.4%), t-cadinol (4.1%), α-humulene oxide (4.0%), caryophyllene oxide (3.6%), bornyl chloride (3.1%), ß-pinene (2.9%) in P. umbellifera. The essential oil of P. salviifolia had a much more complex composition which was dominated by 4-terpineol (13.4%), α-cadinol (5.7%), 6-epi-shyobunol (5.2%), γ-terpinene (5.0%), δ-cadinene (4.4%), α-terpinene (3.5%).

À côté calibration - Making optimal use of time and space in quantitative high performance thin layer chromatography.

Quantitative High Performance Thin Layer Chromatography (HPTLC) requires the application of several standards to each plate, reducing the number of actual samples that can be analyzed in a single run. Using pure standard compounds and a selective detection method, the standards for quantitation can be applied besides - à côté - the chromatography area. This frees the sample application space to accommodate the maximal number of sample on each plate. Also, analysis time is spent exclusively on samples, drastically shortening the effective analysis time per sample and increasing sample throughput. Using this new calibration approach, the sample capacity of regular HPTLC methods can be increased or their scope be extended by an additional quantitative analysis. As a limitation, changes to the distribution of samples and standards within the plate as well as interferences from matrix compounds must be observed. We demonstrate the feasibility of this method by complementing an HPTLC method with a quantitative analysis of total anthocyanin content in colored wheat varieties. The quantitation was validated and compared to the conventional photometric analysis. As outcome, the additional photometric analysis could be replaced and rendered unnecessary, saving time, effort and equipment. This approach could also be employed to quantify highly retained substances, which are usually inaccessible for quantitative analysis.

Phenolic compounds and antioxidant properties of arabinoxylan hydrolysates from defatted rice bran.

BACKGROUND: The water unextractable arabinoxylans (WUAX) contain beneficial phenolic compounds that can be used for food rather than for animal feed. The antioxidant activities of defatted rice bran obtained by xylanase-aided extraction is reported herein. The chemical and molecular characteristics of extracted fractions were investigated. RESULTS: The WUAX hydrolysate precipitated by 0-60% ethanol (F60), 60-90% ethanol (F6090), and more than 90% ethanol (F90) had decreased molar masses with increasing ethanol concentration. The fractions of interest, F60 and F6090, contained 75% arabinoxylans with ferulic acid as the major bound phenolic acid, followed by p-coumaric acid. According to chemical-based antioxidant assays F60 and F6090 exhibited higher diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and ferric iron reducing ability than F90 which contained minor contents of small sugars and free phenolic acids. In cell-based antioxidant assays, using the fluorescent 2',7'-dichlorofluorescein diacetate probe, all three fractions were potent intracellular scavengers. CONCLUSION: The high molar mass of WUAX hydrolysates with high amount of bound phenolics contributes to the chemical-based antioxidant activity. All fractions of WUAX hydrolysates showed high potent intracellular scavenging activity regardless of molar mass, content and the component of bound phenolics. © 2017 Society of Chemical Industry.