Adulteration of apple with pear juice: emphasis on major carbohydrates, proline, and arbutin

Detection of juice-to-juice adulteration based on chemical composition studies is a common method used by government regulatory agencies and food companies. This study investigated the use of major carbohydrate (fructose, glucose and sucrose), polyol (sorbitol), proline, and phenolic profiles as indicators of pear adulteration of apple juice (PAAJ). For this work, a total of 105 authentic apple juice samples from 13 countries and 27 authentic pear juice samples from 5 countries were analyzed. Because the major carbohydrate ranges for these juices showed significant overlap their use as markers for PAAJ detection would be very limited. It was found that sorbitol and proline means for apple and pear were significantly different; however, their broad natural ranges would afford PAAJ at levels up to 30% without detection. In addition, careful selection of the pear juice used as the adulterant would further limit the usefulness of these markers for PAAJ detection. Arbutin was conclusively identified as a marker for pear juice on the basis of its presence in all 27 authentic pear samples and its absence (<0.5 microg/mL) in all 105 apple juice samples analyzed in this study. The application of the developed HPLC-PDA method for arbutin analysis to detect PAAJ at levels as low as 2% (v/v) was demonstrated. A confirmation method for the presence of arbutin in pure pear juice and apple adulterated with pear juice was introduced on the basis of the hydrolysis of arbutin to hydroquinone employing beta-glucosidase, with reactant and product monitoring by HPLC-PDA.     

A new methodology based on GC-MS to detect honey adulteration with commercial syrups

Honey adulterations can be carried out by addition of inexpensive sugar syrups, such as high fructose corn syrup (HFCS) and inverted syrup (IS). Carbohydrate composition of 20 honey samples (16 nectar and 4 honeydew honeys) and 6 syrups has been studied by GC and GC-MS in order to detect differences between both sample groups. The presence of difructose anhydrides (DFAs) in these syrups is described for the first time in this paper; their proportions were dependent on the syrup type considered. As these compounds were not detected in any of the 20 honey samples analyzed, their presence in honey is proposed as a marker of adulteration. Detection of honey adulteration with HFCS and IS requires a previous enrichment step to remove major sugars (monosaccharides) and to preconcentrate DFAs. A new methodology based on yeast (Saccharomyces cerevisiae) treatment has been developed to allow the detection of DFAs in adulterated honeys in concentrations as low as 5% (w/w).     

Improved characterization of the botanical origin of sugar by carbon-13 SNIF-NMR applied to ethanol

Until now, no analytical method, not even isotopic ones, had been able to differentiate between sugars coming from C4-metabolism plants (cane, maize, etc.) and some crassulacean acid metabolism plants (e.g., pineapple, agave) because in both cases the isotope distributions of the overall carbon-13/carbon-12 and site-specific deuterium/hydrogen isotope ratios are very similar. Following recent advances in the field of quantitative isotopic carbon-13 NMR measurements, a procedure for the analysis of the positional carbon-13/carbon-12 isotope ratios of ethanol derived from the sugars of pineapples and agave using the site-specific natural isotopic fractionation-nuclear magnetic resonance (SNIF-NMR) method is presented. It is shown that reproducible results can be obtained when appropriate analytical conditions are used. When applied to pineapple juice, this new method demonstrates a unique ability to detect cane and maize sugar, which are major potential adulterants, with a detection limit in the order of 15% of the total sugars, which provides an efficient mean of controlling the authenticity of juices made from this specific fruit. When applied to tequila products, this new method demonstrates a unique ability to unambiguously differentiate authentic 100% agave tequila, as well as misto tequila (made from at least 51% agave), from products made from a larger proportion of cane or maize sugar and therefore not complying with the legal definition of tequila.     

Mass spectrometric detection and formation of D-amino acids in processed plant saps, syrups, and fruit juice concentrates

Liquid and syrupy dietary saps and juices of plant origin, characterized by the presence of large quantities of saccharides (glucose, fructose, or sucrose) and containing amino acids, were analyzed for the presence of D-amino acids using enantioselective gas chromatography-mass spectrometry. D-amino acids were detected in processed saps and juices of trees (maple, palm, birch), fruits (grape, apple, pear, pomegranate, date), and various other plants (agave, beetroot, sugar cane, carob). D-Ala was detected in all plant products and amounted to approximately 34% D-Ala (relative to L-Ala + D-Ala) in Canadian maple syrups, to approximately 13% in palm saps, and to 48 and 13% D-Ala, respectively, in concentrated grape juices (Spanish Arrope and Turkish Pekmez). Varying amounts and kinds of other D-amino acids were also detected. To test the hypothesis that racemization, that is, partial conversion of L-amino acids into their corresponding D-enantiomers, occurs at reversible stages of the Maillard reaction, the Amadori compound fructose-L-phenylalanine was synthesized. On heating at 200 degrees C for 5 (20) min, release of 10.8% (24.2%) D-Phe was detected. From the data it is concluded that the Amadori compounds formed in the course of the Maillard reaction are pecursors of D-amino acids in foodstuffs.     

Synthesis of oligosaccharides derived from lactulose and pectinex ultra SP-L

The beta-galactosidase activity of the commercial enzymatic preparation Pectinex Ultra SP-L derived from Aspergillus aculeatus has been used to hydrolyze and transgalactosylate the prebiotic carbohydrate lactulose. During this reaction, new oligosaccharides derived from lactulose have been detected by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The presence of the trisaccharide 6'-galactosyl-lactulose, the major compound formed, has been confirmed by NMR. In addition, disaccharides and other oligosaccharides with higher retention times have been also detected. The effect of transgalactosylation conditions such as time, temperature, pH, and initial lactulose and enzyme concentrations, as well as product inhibition on oligosaccharide synthesis, has been studied. The optimal conditions for the formation of tri and higher oligosaccharides were 60 degrees C, pH 6.5, 450 g/L lactulose, 16 units/mL of enzyme, and 7 h of reaction. Selective formation of disaccharides was achieved under the same conditions with the exception of pH (4.5). The present work provides additional knowledge on the synthesis of new oligosaccharides with potential prebiotic properties.     

Melanoidins from coffee infusions. Fractionation, chemical characterization, and effect of the degree of roast

A method involving fractionation in ethanol aqueous solutions, anion exchange chromatography, and immobilized copper chelating chromatography was developed to obtain high molecular weight anionic melanoidin populations from coffee infusions. Six anionic fractions with different physicochemical properties (ethanol solubility and chelating ability) and chemical composition regarding carbohydrate as well as protein nature and content were isolated. Fractions with similar chemical composition were obtained for light-, medium-, and dark-roasted coffee infusions. These melanoidin fractions accounted for 30-33% of the cold-water soluble high molecular weight material, independently of the degree of roast in coffee. The nature and abundance of the different polysaccharides in each fraction were dependent on their ethanol solubility. The 50% ethanol insoluble melanoidin populations contained mostly galactomannan-like carbohydrates, and the fractions obtained with 75% ethanol contained mostly arabinogalactan-like carbohydrates. The melanoidin populations with chelating properties presented significantly lower carbohydrate content and, from these, the 75% ethanol soluble fractions were almost devoid of carbohydrate material. The results obtained suggest that the chelating ability of these coffee melanoidins is modulated by their carbohydrates.     

Gas chromatographic determination of formaldehyde in maple syrup as 2,4-dinitrophenylhydrazone derivative

A procedure is presented for the quantitative extraction and determination of formaldehyde in maple sap and syrup. The method is based on the reaction between formaldehyde and 2,4-dinitrophenylhydrazine and determination of the derivative by gas chromatography. The procedure was used to evaluate formaldehyde in saps and syrups of paraformaldehyde implanted trees. Average recoveries were 101.5 +/- 5.7%. The detection limit was 0.078 mg/kg.     

Comparison of analytical methodologies based on 1h and 31p NMR spectroscopy with conventional methods of analysis for the determination of some olive oil constituents

The present study was designed to assess the agreement between analytical methodologies based on 1H and 31P NMR spectroscopy and conventional analytical methods (titration, gas chromatography, and high performance liquid chromatography) for measuring certain minor and major constituents (free acidity, fatty acids, iodine value, and phenolic compounds) of olive oil. The standard deviations of the NMR method were comparable to those of the conventional methods, except perhaps those of the total hydroxytyrosol and total tyrosol. Linear regression analyses showed strong correlations between NMR and conventional methods for free acidity, total hydroxytyrosol, total tyrosol, total diacylglycerols, (+)-pinoresinol, (+)-1-acetoxypinoresinol, and apigenin; good correlations for linoleic acid, free hydroxytyrosol, and free tyrosol; and weak correlations for oleic acid, linolenic acid, saturated fatty acids, and luteolin. Furthermore, a method comparison study was conducted and the agreement between NMR and conventional methods was evaluated by using the Bland and Altman statistical analysis. The distribution of the data points in the bias plot showed that 96.4% and 100% of the measurements of free acidity and iodine value, respectively, were within the limits of agreement of the two methods. For the remaining constituents of olive oil, the percentage of measurements, located within the limits of agreement, ranged from 94% to 98.5%.     

A strategy for annotating the human milk glycome

Oligosaccharides in human milk represent a group of bioactive molecules that have evolved to be an abundant and diverse component of human milk, even though they have no direct nutritive value to the infant. A recent hypothesis proposes that they could be substrates for the development of the intestinal microflora and the mucosal immune system. The inability to determine the exact composition of these oligosaccharides limits research and the ability to understand their biological functions. Oligosaccharides isolated from the lipids and proteins of individual human milk samples were analyzed by a combination of techniques including microchip liquid chromatography mass spectrometry (HPLC-Chip/MS) and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FT ICR MS). Accurate mass measurements obtained using an orthogonal time-of-flight (o-TOF) mass spectrometry provided oligosaccharide composition for approximately 200 individual molecular species. Comparison of HPLC-Chip/MS profiles from five different women revealed variations in milk oligosaccharide compositions. HPLC-Chip/MS profiling provides a method for routinely identifying milk oligosaccharides. Tandem MS in combination with exoglycosidase digestion provides unambiguous differentiation of structural isomers.     

Ingredient Characterization and Hardening of High‐Protein Food Bars: an NMR State Diagram Approach

A second‐degree simplex lattice mixture design was used to study the effects of soy, dairy, and soy‐dairy blends of powdered proteins in three high‐protein food bar models (sugar syrup, polyol syrup, and reduced‐sugar syrup). Overall protein performance was evaluated based on textural changes during accelerated storage, bar integrity, and dough stickiness and was a strong function of the syrup model used (R² = 92.33%). Nuclear magnetic resonance (NMR) relaxometry was used to measure relaxation times (T₂, T₂*, and T₁) at 20°C and to create state diagrams (temperature, T₂* curves) for the individual powdered proteins and syrups over a temperature range of –35 to 50°C. Increases in relaxation times for powdered protein samples were indicative of better overall protein performance, whereas increases in relaxation times for syrup samples were associated with increases in moisture content and concentration of polyols. Increases in water activity (a_w) of the bars during accelerated storage suggested an elevated rate of hardening for polyol‐containing bars that was caused by a decrease in the amount of water capable of acting as a plasticizer in the product. Proteins were separated into four types (A, B, C, and D) based on the shape of the state diagram curve. Predicted to be the most stable, type D proteins (SUPRO 313 and SUPRO 430) offered the most versatility and, when blended with other proteins, often induced synergistic softening effects in the nutrition bars which led to an extended product shelf life. The NMR state diagram technique appears to be a valuable tool for predicting overall performance of powdered proteins in sugar‐, polyol‐, and reduced‐sugar syrup based food bars.     

Nature of Soil Acidity in Relation to Properties and Lime Requirement of Some Inceptisols

Some Inceptisols representing the Singla catchment area in Karimgaunge district of Assam, India, were studied for lime requirement as influenced by the nature of soil acidity. The electrostatically bonded (EB)-H+ and EB-Al3+ acidities constituted 33 and 67 percent of exchangeable acidity while EB-H+, EB-Al3+,exchangeable and pH-dependent acidities comprised 6, 14, 20 and 80 percent of total potential acidity. The pH-dependent acidity made a major contribution towards the total potential acidity (67%～84%). Grand mean of lime requirement determined by the laboratory incubation method and estimated by the methods of New Woodruff, Woodruff and Peech as expressed in MgCaCO3 ha-1 was in the order: Woodruff (15.6) ＞ New Woodruff (14.9) ＞ Peech (5.1) ＞ incubation (5.0). Correlations analysis among different forms of acidity and lime requirement methods with selected soil properties showed that pH in three media, namely water, 1 mol L-1 KCl and 0.01 mol L-1 CaCl2, had a significant negative correlation with different forms of acidity and lime requirement methods. Exchangeable Fe and Al showed significant positive correlations with EB-Al3+ acidity, exchangeable acidity, pH-dependent acidity and total potential acidity, and also lime requirement methods. Extractable Al showed positive correlations with different forms of acidity except EB-H+ and EB-Al3+ acidities. The lime requirement by different methods depended upon the extractable aluminium.Significant positive correlations existed between lime requirements and different forms of acidity of the soils except EB-H+ acidity and incubation method. The nature of soil acidity was mostly pH-dependent. Statistically, the Woodruff method did slightly better than the New Woodruff, incubation and Peech methods at estimating lime requirement and hence the Woodruff procedure may be recommended for routine soil testing because of its speed and simplicity.     

Study of influential factors on oligosaccharide formation by fructosyltransferase activity during stachyose hydrolysis by Pectinex ultra SP-L

The influence of reaction conditions for oligosaccharide synthesis from stachyose using a commercial enzymatic preparation from Aspergillus aculeatus (Pectinex Ultra SP-L) was studied. Oligosaccharides were analyzed by gas chromatography with flame ionization detection (GC-FID) and matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS). Galactosyl-melibiose (DP(3)) was synthesized as a result of fructosidase activity, whereas fructosyl-stachyose (DP(5)) and difructosyl-stachyose (DP(6)) were formed as a consequence of the fructosyltransferase activity of Pectinex Ultra SP-L. The optimal reaction conditions for the synthesis of penta- and hexasaccharides were 60 °C, pH 5.5, 600 mg/mL stachyose, and 34 U/mL enzyme. Reaction time played an important role in oligosaccharide mixture composition constituted by 20% DP(5), 0.7% DP(6), 55% stachyose, 21% galactosyl-melibiose, and 1% monosaccharides after 1 h and 16% DP(5), 4% DP(6), 27% stachyose, 44% galactosyl-melibiose, and 2% monosaccharides after 3 h. In conclusion, stachyose could be used as a substrate for the enzymatic synthesis of new oligosaccharides that may open new opportunities in the development of future prebiotics.     

Nature of Acidity in Alfisols,Entisols and Inceptisols in Relation to Soil Properties

Twenty acid soils of West Bengal, India, representing Alfisols, Entisols and Inceptisols orders, were studied to characterize their acidity in relation to physicochemical properties. Total potential acidity (TPA), pH-dependent acidity (PDA), total acidity (TA), hydrolytic acidity (HA) and exchange acidity (EA) ranged from 2.02–6.90, 1.75–6.05, 1.18–2.75, 0.98–1.90 and 0.06–0.85 cmol(p⁺) kg^?1, respectively. Relative order for all forms of acidity was: Entisols>Alfisols>Inceptisols. Average contribution of EA to TPA and TA was 9.7% and 19.7%, respectively, and that of PDA to TPA was 90.1%. Contribution of electrostatically bound H⁺ to EA was highest for Inceptisols followed by Entisols>Alfisols, and reverse was true for electrostatically bound aluminum (Al³⁺). All forms of acidity showed significant positive correlations with organic carbon (C) forms of Al but negative correlations with pH of soil. They also showed significant correlations with each other. Soil pH, organic C and exchangeable Al caused most of the variations in different forms of soil acidity.     

High-performance liquid phase separation of glycosides. 5. Determination of individual glucosinolates in cabbage and rapeseed by laser-induced fluorescene capillary electrophoresis via the enzymatically released isothiocyanate aglycon.

A capillary electrophoresis (CE) method was developed for the profiling and determination of individual glucosinolates (GSs) via their isothiocyanate degradation products upon myrosinase digestion. The resulting isothiocyanates, the structures of which are reflective of the parent GS's, were then converted to their corresponding amines via base hydrolysis or reaction with 1, 2-benzenedithiol. Subsequently, the amines were fluorescently labeled to allow their sensitive detection by laser-induced fluorescence (LIF). The CE method involved the use of in situ charged micelles for the separation of isothiocyanates and their corresponding fluorescently labeled amines by micellar electrokinetic capillary chromatography (MECC). The term "in situ charged micelles" refers to micelles formed by complexing the polar hydroxyl groups of glycosidic surfactants with borate. The MECC method with on-column LIF detection was applied to the determination of GSs in white cabbage, rapeseed leaves, and rapeseed roots.     

Validity of carbohydrate, glycaemic index and glycaemic load data obtained using a semi-quantitative food-frequency questionnaire

OBJECTIVE: To assess the ability of a food-frequency questionnaire (FFQ) to rank Australians according to their intake of total carbohydrate, sugar, starch, fibre, glycaemic index (GI) and glycaemic load (GL). DESIGN: Cross-sectional sample from a population cohort. SETTING: Two postcode areas west of Sydney, Australia. SUBJECTS: From 1992 to 1994, a total of 2868 older Australians provided dietary data using a 145-item Willett-derived FFQ. A representative sub-sample of 78 subjects completed three 4-day weighed food records (WFRs). Pearson and Spearman correlations, Bland-Altman plots and weighted kappa values were calculated. RESULTS: Compared with the WFR, the FFQ provided higher mean estimates of all nutrients except starch and GI. All Pearson and/or Spearman correlations were greater than 0.5, except for GL. For GI, sugar, starch and fibre, the regression lines from the Bland-Altman analysis indicated a non-significant linear trend (P = 0.07, P = 0.36, P = 0.28 and P = 0.10, respectively). For GL and total carbohydrate, however, there was a significant linear trend (P = 0.006 and P < 0.0001, respectively), indicating that as the GL and carbohydrate intake of individuals increased, so did the magnitude of the error between the FFQ and WFR. Weighted kappa values all indicated moderate to good agreement, with the exception of GL which was only fair. The proportions of subjects correctly classified within one quintile for all of the nutrients were over 50% and gross misclassification was low (<10%). CONCLUSION: This FFQ was able to rank individuals according to their intakes of total carbohydrate, sugar, starch, fibre and GI, but not as well for GL.     

Use of differential scanning calorimetry (DSC) as a new technique for detection of adulteration in honeys. 1. Study of adulteration effect on honey thermal behavior.

Differential scanning calorimetry (DSC) was used to study the thermal behavior of authentic honeys (Lavandula, Robinia, and Fir honeys) and industrial sugar syrups. Thermal or thermochemical parameters such as the glass transition temperature (Tg), enthalpies of fusion (DeltaH(fus)), and heat capacity variation (DeltaC(p)) were measured. The syrups and honeys showed significant differences in thermal phenomena, as well as in their amplitude and position on the temperature scale. Results showed good reproducibility of the method for all samples studied. The effect of adulteration of honey with different amounts of syrup (5, 10, 20, 40, and 60%) was investigated. A linear relationship was found between the percentage of added syrup and the glass transition temperature. A similar relationship was obtained from the enthalpy of fusion results in the temperature range of 40-90 degrees C. Under applied conditions, the effects of adulteration of honeys by industrial syrups appeared to be detectable from a level as low as 5%.     

Organic matter and carbohydrate distribution in an orthic humic gleysol

Wet sieving was used to separate an Orthic Humic Gleysol into fractions of seven different particle sizes ranging from < 105 μm to 420. All fractions, including the coarse plant residue and water soluble material, were hydrolysed with H₂SO₄ and analysed for total C and neutral sugar content. Their carbohydrate composition was determined by gas-liquid chromatography of the alditol acetates. The major fraction which consisted of particles < 105 μm was enriched by 30% in both organic matter and carbohydrate content. Although the ratio of carbohydrate to organic matter in the major fraction was the same as in the original soil, it varied considerably in the other fractions. The distribution of monomers among the individual fractions varied in a regular pattern. The relative amount of monomers (hexoses, pentoses and deoxyhexoses) in the major fraction ( < 105 μm, and the water soluble material, were essentially the same as in the original soil. It is concluded that the results of this study support the view that the polysaccharides in soil are of bacterial origin.     

Phytochemical and morphological characterization of hop (Humulus lupulus L.) cones over five developmental stages using high performance liquid chromatography coupled to time-of-flight mass spectrometry, ultrahigh performance liquid chromatography photodiode array detection, and light microscopy techniques

Hop (Humulus lupulus L.) inflorescences, commonly known as "hop cones", are prized for their terpenophenolic contents, used in beer production and, more recently, in biomedical applications. In this study we investigated morphological and phytochemical characteristics of hop cones over five developmental stages, using liquid chromatography coupled to time-of-flight mass spectrometry (LC-TOF-MS), and ultrahigh performance liquid chromatography photodiode array detection (UHPLC-PDA) methods to quantitate 21 polyphenolics and seven terpenophenolics. Additionally, we used light microscopy to correlate phytochemical quantities with changes in the morphology of the cones. Significant increases in terpenophenolics, concomitant with glandular trichome development and associated gross morphological changes, were mapped over development to fluctuations in contents of polyphenolic constituents and their metabolic precursor compounds. The methods reported here can be used for targeted metabolic profiling of flavonoids, phenolic acids, and terpenophenolics in hops, and are applicable to quantitation in other crops.     

Carbohydrates in cyanobacterial soil crusts as a source of carbon in the southwest Kalahari, Botswana

Semiarid ecosystems, also known as drylands, contain small amounts of carbon (C). To date only few studies have evaluated soil C dynamics in these ecosystems. Cyanobacterial soil crusts are considered a major source of organic C in semiarid ecosystems through photosynthesis, increasing soil organic C (SOC) pool as carbohydrates. This study considers cyanobacterial soil crusts as a source of C in the southwest Kalahari. Cyanobacterial soil crusts are well adapted to drylands because extracellular polysaccharide (EPS) secretions provide stabilization of soil and resistance to desiccation. The carbohydrate and chlorophyll a content were evaluated in the upper soil profile on Kalahari Sands and pan soils. Topsoil carbohydrate concentration decreased exponentially with depth. The carbohydrate content produced in cyanobacterial soil crusts in the southwest Kalahari represents up to 75% of the total SOC and is thus an essential component for the fertility of Kalahari Sand soils.     

Kinetic study of the thermal hydrolysis of Agave salmiana for mezcal production

The kinetics of the thermal hydrolysis of the fructans of Agave salmiana were determined during the cooking step of mezcal production in a pilot autoclave. Thermal hydrolysis was achieved at different temperatures and cooking times, ranging from 96 to 116 °C and from 20 to 80 h. A simple kinetic model of the depolymerization of fructans to monomers and other reducing sugars and of the degradation of reducing sugars to furans [principally 5-(hydroxymethyl)furfural, HMF] was developed. From this model, the rate constants of the reactions were calculated, as well as the pre-exponential factors and activation energies of the Arrhenius equation. The model was found to fit the experimental data well. The tradeoff between a maximum fructan hydrolysis and a critical furan concentration in allowing for the best ethanol yield during fermentation was investigated. The results indicated that the thermal hydrolysis of agave was optimal, from the point of view of ethanol yield in the ensuing fermentation, in the temperature range of 106-116 °C and the cooking range time of 6-14 h. The optimal conditions corresponded to a fructan hydrolysis of 80%, producing syrups with furan and reducing sugar concentrations of 1 ± 0.1 and 110 ± 10 g/L, respectively.