Synthesis of carboxymethyl cellulose (CMC) and starch-based hybrids and their applications in flocculation and sizing

Starch and hydrolyzed starches along with carboxymethyl cellulose were independently subjected to cationization by reacting them with N-(3-chloro-2-hydroxypropyl) trimethylammonium chloride (known commercially as Quat 188) in presence of alkaline catalyst. The cationization reaction was carried out under different conditions Optimal reaction conditions could be achieved at Quat 188 (2 mmol), NaOH (1 mmol) at temperature (70 °C) 60 min for CMC or 120 min for starch. The cationic products were characterized by FTIR spectroscopy, and nitrogen percentage. Application of the cationized products as well as their parent materials in textile sizing was also undertaken. Moreover, hybrids of starch-Quat 188 and CMC-Quat 188 were used as flocculants; flocculation was carried at pH 6 using different flocculant dose. Results obtained indicate that the cationic-based hybrids – under investigation display characteristics, which qualify them to function as an excellent sizing and flocculating agents.     

Effects of reaction conditions on the physicochemical properties of cationic starch studied by RSM

Granular cationic starches were prepared in aqueous phase without the addition of swelling inhibiting salts. Response surface methodology (RSM) was performed to analyze the effects of reaction conditions on physicochemical properties of the products. As the reaction time was prolonged from 2 to 5 and to 24 h, the relative contribution of the temperature to degree of substitution (DS) turned from minor to prominent. Good correlations were observed between the DS and the pasting temperature of the 2, 5, and 24 h cationized starches. By contrast, variation in the correlations between DS and the other physicochemical properties, with respect to reaction durations, revealed the processing pattern of cationization in starch granules along extending reaction times. This deduced pattern was confirmed by the granular and molecular characterizations using confocal laser scanning microscopy and high performance anion exchange chromatography, respectively.     

Preparation and characterization of cationic corn starch with a high degree of substitution in dioxane-THF-water media

Cationic corn starch derivatives with a high degree of substitution are prepared in alkaline solution or in mixed media of organic solvent and water with different levels of the cationic reagent, 2,3-epoxypropyltrimethylammonium chloride. The starch cationization yield is investigated, and the results indicate that the degree of substitution (DS) of the samples depends on the reaction conditions and reaction media. The maximum DS values are up to 1.37 in 1,4-dioxane alkaline-aqueous solution. Meanwhile, the structures of the cationic starch derivatives are characterized by elemental analyses, FTIR spectroscopy, X-ray diffraction, and ¹³C NMR spectroscopy, as well as by SEM techniques.     

Optimization of the synthesis of 1-allyloxy-2-hydroxy-propyl-starch through statistical experimental design

The synthesis of 1-allyloxy-2-hydroxy-propyl starches was studied using a statistical experimental design approach. The etherification of two different granular maize starches with allyl glycidyl ether (AGE) in a heterogeneous alkaline suspension was investigated. The optimal reaction conditions were found via experimental design and the obtained response factor, e.g. the degree of substitution (DS) of the starch hydroxyl group, was statistically evaluated. The effects of six process factors on DS, namely the starch concentration, the reaction time, the temperature, and the amount of NaOH, Na₂SO₄ and AGE were investigated. The statistical analysis showed significant impact of the temperature, the amount of NaOH and the amount of AGE on the DS for both starches. Optimum conditions for the highest DS for waxy maize starch were: 0.0166% AGE (based on dry starch (ds)) and 1.0% NaOH (ds) at 34 °C in 4 h; on dent maize starch, these were 0.0099% AGE (ds) and 1.0% NaOH (ds) at 37 °C in 16 h.     

Carboxymethyl Chinese yam starch: synthesis, characterization, and influence of reaction parameters

Carboxymethyl starch (CMS) was obtained as a product of the reaction of starch and monochloroacetic acid (MCA) in the presence of sodium hydroxide. The influence of the molar ratio of NaOH/AGU, the molar ratio of MCA/AGU, the reaction time, reaction temperature, and the water content on the degree of substitution (DS) was studied. The optimal molar ratio of NaOH/AGU and MCA/AGU is 2.4 and 1.0, respectively. Increase of the ratio of NaOH/AGU or MCA/AGU leads to an increase in DS, but only to certain extent. The highest values of the DS were obtained when the carboxymethylation was performed at 60 °C for 2.5 h. The water content in the reaction media ethanol was optimal at 20% (v/v). The scanning electron micrographs (SEMs) revealed that the carboxymethylation affected the structural arrangement of the starch and caused granular disintegration. The particle size distribution (PSD) also displayed that the average particle diameter increased greatly after modification from 37.37 μm to 72.88 μm. Wide angle X-ray diffractometry (XRD) revealed that starch crystallinity was obviously reduced after carboxymethylation. The new bands at 1600 cm⁻¹ and 1426 cm⁻¹ in FT-IR indicated that the starch granules were substituted.     

Functional properties of pregelatinized and cross-linked cassava starch obtained by extrusion with sodium trimetaphosphate

Cassava starch was cross-linked with sodium trimetaphosphate (STMP) on a Cerealtec single-screw extruder at different extrusion temperatures and concentrations of STMP and NaOH. The effect of variables on functional properties of the products was analysed by the response–surface methodology. The degree of substitution (DS) was influenced by NaOH and phosphorus level, and increased when their concentration increased. Extrusion temperature affected water absorption, cold viscosity and gel characteristics. The introduction of phosphate groups by cross-linking, with maximum DS of 1.5×10⁻⁴, increased the gel strength, water absorption index, resistance to high temperature and shear, and decreased gel cohesiveness, starch clarity and water solubility index. The products had different DS and degree of gelatinization and thus can be applied in several kinds of foods.     

Preparation and Application of Carboxymethyl Yam (Dioscorea esculenta) Starch

Yam (Dioscorea esculenta) starch was modified by carboxymethylation. The effect of reaction parameters, amount of sodium hydroxide (NaOH), amount of sodium monochloroacetate (SMCA), and reaction time on the degree of substitution (DS) of carboxymethyl yam starch (CMS), was studied using the Box–Behnken experimental design. Physicochemical and potency to be a tablet disintegrant of CMS were evaluated. CMS with DS in the range of 0.08–0.19 were obtained. The results from regression analysis indicated that the most important factor in controlling DS was the amount of NaOH followed by SMCA content and reaction time. However, high concentration of NaOH and SMCA lowered the DS. The optimal conditions to achieve the highest DS (0.19) were found to be at molar ratios of NaOH and SMCA to anhydroglucose unit of 1.80 and 2.35, respectively, and with the reaction time of 4.8 h. The swelling power and viscosity of CMS increased with an increase in the degree of modification. CMS showed satisfying tablet disintegrant properties. The tablets containing 1.0–4.0 % CMS disintegrated faster than 5 min. Hence carboxymethyl yam starch can be used as an excellent tablet disintegrant in low concentration.     

Analysis of solvent-free ethyl oleate enzymatic synthesis at equilibrium conditions

This work reports experimental equilibrium data for the esterification of pure oleic acid and a fatty acid mixture with ethanol, using an immobilized Candida antarctica B lipase as catalyst. Reactions are performed in a solvent-free system, containing a mixture of substrates and different amounts of distilled water. According to the initial amount of water and the extent of the reaction, one or two liquid phases are present. Therefore, when the equilibrium is achieved, the liquid–liquid and chemical reaction equilibria have to be simultaneously satisfied.

Several reports dealing with enzymatic reactions performed in two-phase systems have found that the value of the reaction equilibrium constant calculated from overall experimental concentrations varies not only with temperature but also with substrate ratio and water content. Although this approach is a valuable way to explore equilibrium shifts in biphasic systems, it is limited to ideal systems with constant partition coefficients. The aim of this work is to consider the biphasic nature of the reactive mixture through a computational procedure that simultaneously takes into account liquid–liquid and reaction equilibria. This approach enables the determination of a classical temperature-dependent thermodynamic equilibrium constant, which accurately fits experimental equilibrium conversions over a wide range of operating conditions.     

Properties of new alkaline maltohexaose-forming amylases

Summary Three alkaline amylases have been newly discovered in a culture medium of an alkalophilic Bacillus sp. H-167 isolated from soils. These amylases produced maltohexaose as the main product from starch. All three amylases were purified to give a single band on disc electrophoresis. They had similar properties except for molecular weight (MW) and isoelectric point (pI): optimum pH, 10.5; optimum temperature, 60°C; pH stability, 7–12; heat stability, 50–55°C; MW, 59000–80000; pI, 3.5–4.3. Metal ions such as Hg²⁺, Zn²⁺, Pb²⁺, Co²⁺ and Ni²⁺ inhibited the enzyme activity. All the enzymes hydrolyzed starch to produce preferentially maltohexaose, rather than maltose and maltotetraose, in an early stage of the reaction. The yield of maltohexaose reached 25%–30% from soluble starch.     

Synthesis of superabsorbent from carbohydrate waste

A mixture of acrylamide (AM) and acrylic acid (AA) monomers were grafted on germinated gelatinized wheat starch using potassium persulfate (KPS) as an initiator to yield superabsorbent. The effects of different parameters such as time, temperature, monomer feed (AM:AA), starch: monomers ratio and initiator concentration on graft add-on were studied, and the optimized values were found to be 2 h, 60 °C, 1:1 (w/w), 1:1(w/w) & 1% (with respect to starch powder), respectively. The product so formed was saponified with 0.1N NaOH, dried and finely powdered sample was characterized using FT-IR, TGA. Product showed maximum absorbency of 150 g/g.     

Synthesis, analysis and reduction of 2-nitropropyl starch

Granular 2-nitropropyl potato starch was synthesized by reaction with 2-nitropropyl acetate in an aqueous suspension. Nitroalkylation occurs preferentially with the amylose fraction of potato starch, as was confirmed by leaching experiments and digestion of the modified starch with alpha-amylase. The 2-nitropropyl substituent is a mixture of the nitroalkane and nitronic acid tautomer. Some grafting occurs and to a lesser extent additional reactions (formation of carbonyls and oximes) of the nitro group take place. After catalytic hydrogenation of water soluble 2-nitropropyl starch only a small amount of the nitro functionality was reduced to the corresponding amine. Reduction of granular 2-nitropropyl starch with sodium dithionite did not go to completion and led to a complex mixture of starting material, several intermediates and side products (for example sulfamates).     

Quantitative characteristic of the catalytic properties and microstructure of cross-linked enzyme aggregates of penicillin acylase

The microstructure and the catalytic properties of cross-linked enzyme aggregates (CLEA) of penicillin acylase (PA) obtained under different conditions were investigated. The period of time left between the enzyme precipitation and the cross-linking step was found to influence the structural organization of the resulting enzyme preparation. Confocal fluorescent microscopy of the so-called “fresh” and “mature” CLEAs PA allowed to estimate the “characteristic” diameter of CLEA PA particles, which appeared to be about 1.6 μm, and revealed that the “mature” type was composed of relatively big particles as compared to the “fresh” type. Complementary kinetic studies showed that the “mature” CLEA PA were more effective in both hydrolytic and synthetic reactions. It was suggested that the aggregate size might regulate the extent of covalent modification of PA and thereby influence the catalytic properties of CLEA.     

Resistant glutarate starch from adlay: Preparation and properties

Reaction conditions were optimized to increase the content of resistant starch in adlay starch using esterification with glutaric acid, and the physicochemical properties of the prepared glutarate starches were investigated. Different amounts of glutaric acid (0.1–0.5 g/g starch, dry weight basis) were reacted with adlay starch at various temperatures (70–130 °C) and reaction times (3–9 h). The resistant starch levels increased with increased glutaric acid content, reaction temperature, and reaction time. The color difference was mainly affected by reaction time. The highest resistant starch content (RS 66%) was obtained using conditions of 0.4 g glutaric acid/g starch, 115 °C, and 7.5 h, with a color difference of 10.24. After digestion with α-amylase and amyloglucosidase, the water-soluble fraction of glutarate starch had more oligosaccharides than high-amylose maize starch (RS 43%). FT-IR and solid-state NMR detected carbonyl groups in the glutarate starch, indicating the formation of cross-linkages through esterification. The granular structure of the glutarate starches was not destroyed and they retained birefringence. After heating with an excess of water, the granules kept their shape but lost their birefringence. The glutarate starches had low solubility in both cold and hot water, and the resistant starch contents were unchanged after heating due to the restriction of swelling by cross-linking. The glutarate starches had a similar chain-length distribution to raw starch, indicating that acid hydrolysis took place at branching points in the amorphous region. Furthermore, the glutarate starches possessed a weaker crystalline region, more diverse double helical chains, and lower enthalpy than raw starch.     

Preparation of cassava starch grafted with polystyrene by suspension polymerization

Cassava starch grafted with polystyrene (PS-g-starch) copolymer was synthesized via free-radical polymerization of styrene by using suspension polymerization technique. Potassium persulfate (PPS) was used as an initiator and water was used as a medium. The graft copolymer was characterized by Fourier transform infrared spectroscopy, differential scanning calorimetry, thermal gravimetric analysis, X-ray diffraction and scanning electron microscopy. The sub-micron spherical beads of PS were observed on the surface of starch granules. SEM micrographs showed porous patches of PS adhering on the starch granules after Soxhlet extraction. FTIR spectra also indicated the presence of PS-g-starch copolymer. XRD analysis exhibited insignificant changes in crystalline structure and degree of crystallinity. The effects of starch:styrene weight ratio, amount of PPS, reaction time and reaction temperature on the percentage of grafting – G (%), were investigated. G (%) increased with increasing starch content. Other variables showed their own individual optimal values. The optimum condition yielding 31.47% of G (%) was derived when the component ratio was 1:3 and reaction temperature and time were 50 °C and 2 h, respectively. Graft copolymerization did not change granular shape and crystallinity of starch. This study demonstrated the capability of polymerization of styrene monomer on the granular starch without emulsifier and the synthesis of graft copolymer without gelatinization of starch.     

Formamide denaturation of chromosomal DNA for in situ hybridization and C-band preparation in the guinea pig, Cavia porcellus

Cytological preparations were incubated in 0.07 N NaOH at room temperature or 90% formamide (final salt concentration 2 × SSC) at either 65 °C or 37 °C for 2.5 h to denature guinea pig chromosomes. Chromosomes treated with NaOH or formamide at 65 °C showed a large amount of DNA loss, while chromosomes treated with formamide at 37 °C showed little or no DNA loss. Repeated sequences were isolated from guinea pig DNA and [³H]cRNA was transcribed with Escherichia coli RNA polymerase for in situ hybridization. Localization of the [³H]cRNA occurred in the centromeric regions and C-band positive short arms of almost all of the chromosomes in the NaOH preparations. Chromosomes treated with formamide at 65 °C showed the same grain distribution with a decrease in the number of grains/cluster. Slides incubated in formamide at 37 °C showed localization in only a few chromosomes and the number of grains/cluster was greatly diminished. Thermal denaturation of isolated chromatin indicated that incubation of chromosomes in formamide at 37 °C did not fully denature the DNA. C-bands could be induced by treating slides in formamide at either 65 °C or 37 °C when followed by a “reassociation” in 2 × SSC at 65 °C for 16 h. If the “reassociation” step was omitted, C-bands were found in the 65 °C formamide slides but not the 37 °C formamide slides.     

Preparation of cationic cotton with two-bath pad-bake process and its application in salt-free dyeing

Cationic cotton was prepared by a designed two-bath pad-bake process with 3-chloro-2-hydroxypropyltrimethylammonium chloride as cationizing reagent to realize recycle utilization of the reagent and continuous processing of cationization. Experiments showed that 8.0% (o.w.bath) of the reagent, 1:1 of molar ratio of sodium hydroxide to the reagent, 60 °C and 6 min of baking temperature and time were selected for cationization and the obtained cationic cotton was suitable for application in salt-free reactive dyeing. The structures of both the untreated and cationic fibers were investigated by X-ray diffraction and scanning electronic microscopy. Higher dye utilization and color yields could be realized on the cationic cotton than that on the untreated one in the conventional dyeing. Levelness dyeing and good fastness properties of the dyes on the cationic fabrics were obtained. Besides, colorimetric properties and mechanical strength of the dyed fabrics were both evaluated to show applicability of this preparation process of cationic cotton.     

Ester-exchange reaction between triglycerides with polyethylene glycol-modified lipase

Summary Polyethylene glycol-modified lipase efficiently catalyzed esterexchange reaction between trilaurin and triolein in the straight hydrophobic substrates. Dilauroyl-monooleoyl-glycerol and monolauroyl-dioleoyl-glycerol were formed from two triglyceride-substrates, trilaurin and triolein, in the presence of the modified lipase at 58°C. Consequently, the melting temperature of the mixture of two substrates was decreased from 33–36°C to 11–13°C. Similar ester-exchange reaction took place between fat and oil composed of triglycerides, accompanied by the decrease in the melting temperature of the reaction mixture.     

Bioanalysis of aplidine, a new marine antitumoral depsipeptide, in plasma by high-performance liquid chromatography after derivatization with trans-4′-hydrazino-2-stilbazole

A sensitive bio-analytical assay in plasma of the depsipeptide aplidine is reported, based on reversed-phase liquid chromatography and fluorescence detection of the trans-4′-hydrazino-2-stilbazole (4′H2S) derivative of the analyte. At ambient temperature, two conformations of the depsipeptide are observed in solution due to cis–trans isomerism at the proline–pyruvoyl peptide bond. Aplidine is isolated from the matrix by solid-phase extraction on an octadecyl modified silica stationary phase. After evaporation of the acetone eluate, a derivatization with 4′H2S is performed in a water–acetonitrile mixture at pH 4. The reaction mixture is injected directly into the chromatograph and the analyte is quantified by fluorescence detection at 410 and 560 nm for excitation and emission, respectively. The method has been validated in the 2–100 ng/ml-range, 2 ng/ml being the lower limit of quantification. Precision and accuracy both meet the current requirements for a bioanalytical assay. The identity of the 4′H2S reaction products of aplidine have been confirmed by mass spectrometric analysis. Finally, the method has been employed for a pilot pharmacokinetic study of aplidine in mice which demonstrated its usefulness for pharmacological research.     

Preparation of clear noodles with mixtures of tapioca and high-amylose starches

Ways to simulate the making of clear noodles from mung bran starch were investigated by studying the molecular structures of mung bean and tapioca starches. Scanning electron micrographs showed that tapioca starch granules were smaller than those of mung bean starch. X-ray diffraction patterns of mung bean and tapioca starch were A- and C_A-patterns, respectively. Iodine affinity studies indicated that mung bean starch contained 37% of apparent amylose and tapioca starch contained 24%. Gel permeation chromatograms showed that mung bean amylopectin had longer peak chain-length of long-branch chains (DP 40) than that of tapioca starch (DP 35) but shorter peak chain-length of short-branch chains (DP 16) than that of tapioca starch (DP 21). P-31 n.m.r. spectroscopy showed that both starches contained phosphate monoesters, but only mung bean starch contained phospholipids. Physical properties, including pasting viscosity, gel strength, and thermal properties (gelatinization), were determined. The results of the molecular structure study and physical properties were used to develop acceptable products using mixtures of cross-linked tapioca and high-amylose maize starches. Tapioca starch was cross-linked by sodium trimetaphosphate (STMP) with various reaction times, pH values, and temperatures. The correlation between those parameters and the pasting viscosity were studied using a visco/amylograph. Starches, cross-linked with 0.1% STMP, pH 11.0, 3.5 h reaction time at 25, 35, and 45°C (reaction temperature), were used for making noodles. High-amylose maize starch (70% amylose) was mixed at varying ratios (9, 13, 17, 28, 37, and 44%) with the cross-linked tapioca starches. Analysis of the noodles included: tensile strength, water absorption, and soluble loss. Noodle sensory properties were evaluated using trained panelists. Noodles made from a mixture of cross-linked tapioca starch and 17% of a high-amylose starch were comparable to the clear noodles made from mung bean starch.     

Effects of protein on crosslinking of normal maize, waxy maize, and potato starches

Channels of maize starch granules are lined with proteins and phospholipids. Therefore, when they are treated with reagents that react at or near the surfaces of channels, three types of crosslinks could be produced: protein–protein, protein–starch, starch–starch. To determine which of these may be occurring and the effect(s) of channel proteins (and their removal) on crosslinking, normal and waxy maize starches were treated with a proteinase (thermolysin, which is known to remove protein from channels) before and after crosslinking, and the properties of the products were compared to those of a control (crosslinking without proteinase treatment). After establishing that treatment of starch with thermolysin alone had no effect on the RVA trace, three reaction sequences were used: crosslinking alone (CL), proteinase treatment before crosslinking (Enz-CL), proteinase treatment after crosslinking (CL-Enz). Two crosslinking reagents were used: phosphoryl chloride (POCl₃), which is known to react at or near channel surfaces; STMP, which is believed to react throughout the granule matrix. Three concentrations of POCl₃ (based on the weight of starch) were used. For both normal maize starch (NMS) and waxy maize starch (WMS) reacted with POCl₃, the trends were generally the same, with apparent relative degrees of crosslinking indicated to be CL-Enz = CL > Enz-CL, but the effects were greater with NMS and there were differences when different concentrations of reagent were used. The basic trends were the same when potato starch was used in the same experiments. Crosslinking with STMP was done both in the presence and the absence of sodium sulfate (SS). Both with and without SS and with both NMS and WMS, the order of indicated crosslinking was generally the same as found after reaction with POCl₃, with the indicated swelling inhibition being greater when SS was present in the reaction mixture. Examination of the maize starches with a protein stain indicated that channel protein was removed by treatment with thermolysin when the proteinase treatment occurred before crosslinking with either POCl₃ or STMP, but only incompletely or not at all if the treatment with the proteinase occurred after crosslinking. Because the crosslinking reactions were less effective when the protein was removed, the results are tentatively interpreted as indicating that they involved protein molecules, although there may not be a direct relationship.