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Stuart Chalk, Ph.D.
Department of Chemistry
University of North Florida
Phone: 1-904-620-1938
Fax: 1-904-620-3535
Email: schalk@unf.edu
Website: @unf

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Sucrose

  • IUPAC Name: (2R,3R,4S,5S,6R)-2-[(2S,3S,4S,5R)-3,4-dihydroxy-2,5-bis(hydroxymethyl)oxolan-2-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol
  • Molecular Formula: C12H22O11
  • CAS Registry Number: 57-50-1
  • InChI: InChI=1S/C12H22O11/c13-1-4-6(16)8(18)9(19)11(21-4)23-12(3-15)10(20)7(17)5(2-14)22-12/h4-11,13-20H,1-3H2/t4-,5-,6-,7-,8+,9-,10+,11-,12+/m1/s1
  • InChI Key: CZMRCDWAGMRECN-UGDNZRGBSA-N

@ ChemSpider@ NIST@ PubChem

Citations 62

"Combination Of Flow Injection With Capillary Electrophoresis - Part 7. Microchip Capillary Electrophoresis System With Flow Injection Sample Introduction And Amperometric Detection"
Anal. Chim. Acta 2000 Volume 422, Issue 1 Pages 71-79
Chong-Gang Fu and Zhao-Lun Fang

Abstract: A microchip-based capillary electrophoresis (CE) system with amperometric detection, combined with flow injection (FI) sample introduction, was constructed from components readily available in the analytical laboratory. An H-channel configuration was used with separation capillary positioned between two tubular side-arms, and a falling-drop interface connected to one side-arm was developed to achieve electrical isolation between the FI and CE systems. End-column amperometric detection was accomplished with separation voltage decoupled from the detection system, employing a microdisk working electrode positioned immediately outside the capillary outlet in the other side-arm, which functioned as a large-volume reservoir. Sample dispersion in the FI system and the FI-CE interface was minimized by intercalating the sample zone between two air segments. Performance of the FI-CE amperometric system was demonstrated by separation of sugars. Separation of sucrose and glucose was performed with a 5 cm, 25 µm i.d. capillary with 1.7 kV separation voltage in about 60 s (40 µm plate height for glucose), achieving a sampling frequency of over 65 h-1. Responses for sucrose and glucose were linear in the range of 10^-1000 µM with sensitivities of 0.011 and 0.025 nA µM-1. Detection limits (S/N = 3) were 2 µM for sucrose and 1 µM for glucose. Peak height precisions were 2.1 and 2.4% RSD (n = 9) for sucrose and glucose, respectively.
Amperometry Electrophoresis Microfluidic Interface

"Sequential Injection Fourier Transform Infrared Spectroscopy For The Simultaneous Determination Of Organic Acids And Sugars In Soft Drinks Employing Automated Solid Phase Extraction"
Anal. Chim. Acta 2000 Volume 422, Issue 1 Pages 63-69
Hai LeThanh and Bernhard Lendl

Abstract: A fully automated method for the rapid determination of organic acids (citric-, malic- and tartaric acid) and sugars (glucose, fructose, and sucrose) in soft drinks by sequential injection Fourier transform infrared (FTIR) spectroscopy is presented. A convective interaction media (CIM) disc carrying quaternary amino moieties was added as a solid phase extraction column to the flow system. Upon injection of a sample the organic acids were completely retained on the CIM disc whereas sugars passed to the flow cell. The organic acids were subsequently eluted by injection of an alkaline (pH 8.5) 1 M sodium chloride solution and recorded in their fully deprotonated form as a second flow injection peak. In both cases, the FTIR spectra corresponding to the peak maxima were selected for data evaluation. Two partial least squares models, one for sugars and the other for organic acids, were constructed based on the analysis of standards containing all six analytes. The developed method was applied to natural samples yielding results which were in good agreement with those obtained by an external reference method (enzymatic test kits). Deviations in the results were 3.4. and 4.1% for citric and malic acid and ranged from 4.7-5.1% for the sugars. The developed method is characterized by its short analysis time, experimental simplicity and its potential applications in routine analysis and process control.
Soft drink Spectrophotometry Sequential injection Preconcentration Method comparison Solid phase extraction

"Sequential Flow Injection Analysis Based On Calorimetric Detection"
Thermochim. Acta 1999 Volume 337, Issue 1-2 Pages 27-38
A. Wolf, A. Weber, R. Hüttl, J. Lerchner and G. Wolf

Abstract: A calorimetrically based sequential flow injection method with enzyme catalyzed reactions is proposed and demonstrated. In contrast to multi-detection devices with immobilized enzymes miniaturized flow-through reaction calorimeters are used as sensing device with independent substrate and toggled enzyme solution flows. From the sequence of the heat power signals information about composition of the substrate is obtainable. A reasonable application of the method requires miniaturized calorimeters. Different constructions of flow-through reaction calorimeters based on silicon chips with integrated thermopiles as heat power transducers have been analyzed with respect to their sensitivities and mixing behavior. As an application the analysis of saccharides containing mixtures is discussed.
Calorimetry Microfluidic

"Dual Electrode Signal-subtracted Biosensor For Simultaneous Flow Injection Determination Of Sucrose And Glucose"
Anal. Chim. Acta 1999 Volume 380, Issue 1 Pages 7-15
Werasak Surareungchai, Supinda Worasing, Pornpimol Sritongkum, Morakot Tanticharoen and Krissanapong Kirtikara

Abstract: A multi-enzyme electrode obtained by a two-step immobilization of the enzymes glucose oxidase, mutarotase and invertase was developed for the determination of sucrose. Glucose oxidase was entrapped in a poly-1,3-diaminobenzene film on a platinum electrode by electrochemical polymerization and a combination of mutarotase and invertase was cross-linked over the electrode via bovine serum albumin and glutaraldehyde. The sucrose concentration was determined from hydrogen peroxide oxidation at +0.7 V vs. Ag/AgCl. This immobilization method minimized interference from ascorbic acid. A second electrode, for glucose only, was constructed containing inactive invertase. This gave an almost identical glucose response to the sucrose sensor and hence could be used for signal subtraction. In this manner, sucrose could be determined in the presence of glucose at higher concentrations. The influence of enzyme content, pH, temperature, and optimum operational conditions for use in a flow injection analysis (FIA) system were determined. When used in FIA, the sensor responded to sucrose in the range 1-300 mmol l-1. The sensor was stable for 15 h of continuous use (150 assays) and retained 70% activity after 30 days. When used to analyze the sucrose and glucose contents in a number of different soft drinks, the sensor showed good agreement with the standard liquid chromatography (LC) method.
Soft drink Amperometry Immobilized enzyme Optimization Method comparison Interferences

"Computer Monitoring Of Sugars, Acids And Volatile Compounds In Fermentations"
Anal. Chim. Acta 1984 Volume 163, Issue 1 Pages 275-280
J. C. Motte, X. Monseur, M. Termonia, M. Hofman, G. Alaerts, A. De meyer, P. Dourte and J. Walravens

Abstract: A brief account is given of the use of head-space g.c. - computer-assisted m.s. to monitor the formation of ethanol in fermentation media, HPLC with computerized integration to monitor the disappearance of sugars and the formation of acids during fermentation, and flow injection analysis to monitor feed streams of low sugar content.
Fermentation broth Computer

"Simultaneous Determination Of Sucrose And Glucose In Mixtures By Flow Injection Analysis With Immobilized Enzymes"
Anal. Chim. Acta 1985 Volume 171, Issue 1 Pages 185-194
M. Masoom and Alan Townshend

Abstract: Sucrose was determined in a flow injection system by using a manifold comprising columns of β-D-fructofuranosidase - aldose 1-epimerase and glucose oxidase (each immobilized on controlled-pore glass) in sequence, followed by amperometric detection of the H2O2 produced. At pH 6.8, max. sensitivity was achieved, the enzyme maintained activity for ~12 weeks, and the detection limit was 10 µM-sucrose. Incorporation of a controlled by-pass around the first column allowed one sample to traverse both columns, thereby allowing the sequential determination of sucrose and glucose (0.1 to 10 mM), while the next sample passed through only the glucose oxidase column; accurate determinations were possible within 25 s. The simultaneous determination of sucrose and glucose was also possible (in 2 min) by means of a single injection, which was split in the apparatus, thus allowing a sucrose - glucose and a glucose peak to be obtained in sequence. For five samples, the coefficient of variation was 1.8% for the combined peak and 2.9% for the glucose peak.
Amperometry Immobilized enzyme Controlled pore glass Simultaneous analysis

"Determination Of Sucrose In The Presence Of Glucose In A Flow Injection System With Immobilized Multi-enzyme Reactors"
Anal. Chim. Acta 1986 Volume 179, Issue 1 Pages 203-208
Bo Olsson, Berit StÅlbom and Gillis Johansson

Abstract: Sucrose is determined in a packed-bed reactor by its reactions with co-immobilized β-fructofuranosidase, aldose-1-epimerase(I) and glucose oxidase(II). Glucose in the sample is decomposed in a pre-reactor containing co-immobilized I, II and catalase. The H2O2 produced is measured at 514 nm after reaction with a reagent comprising 20 mM 3,5-dichloro-2-hydroxybenzenesulfonate, 1 mM 4-aminoantipyrine and 1 mM EDTA in 0.1 M citrate buffer, pH 6.0. The response is rectilinear from the detection limit, 0.1 µM, up to 0.5 mM. The coefficient of variation at the 0.2 mM level is 0.3% (n = 20).
Spectrophotometry Immobilized enzyme Reactor

"Determination Of Sucrose In Sugar-cane Juice And Molasses By Flow Injection Spectrophotometry"
Anal. Chim. Acta 1988 Volume 204, Issue 1-2 Pages 259-270
E. A. G. Zagatto, I. L. Mattos and A. O. Jacintho

Abstract: The diluted and filtered sugar cane juice or molasses is introduced into a flow injection analyzer. with two merging streams, producing two sample zones. One zone is fed directly to the confluence of the streams but the other first flows through a heated coil where partial and reproducible sucrose (I) inversion is achieved at controlled pH and temperature At the confluence point a buffered IO4- reagent is added to oxidize I. The consumption of IO4- is measured spectrophotometrically as a transient lowering of the iodine concentration, produced by reaction of IO4- with I-. The two zones are fed sequentially to a flow cell where two peaks are recorded. The I concentration. is proportional to the difference in peak heights. About 30 samples h-1 of sugar-cane juice can be analyzed. The coefficient of variation was 0.51% for a I concentration. of 13.66% in cane juice (n = 7). Similar precision was obtained with the modified system for molasses (20 samples h-1).
Juice Vegetable Spectrophotometry Indirect Merging zones Optimization Heated reaction

"Flow Injection Determination Of Sugars With Immobilized Enzyme Reactors And Chemiluminescence Detection"
Anal. Chim. Acta 1988 Volume 205, Issue 1-2 Pages 195-205
Cathy A. (Koerner) Swindlehurst and Timothy A. Nieman

Abstract: In the determination of glucose, the sample (80 µL) was injected into a carrier stream of 1 mM phosphate buffer (pH 6.5), which was passed through a reactor (5.6 cm x 3 mm) containing glucose oxidase and aldose 1-epimerase, immobilized on controlled-pore glass, and the H2O2 produced was determined via its chemiluminescence reaction with luminol in the presence of peroxidase at pH 11.6. Sucrose, maltose, lactose and fructose were determined similarly after their enzymatic conversion into glucose (with β-fructofuranosidase and aldose 1-epimerase, glucan 1,4-α-glucosidase, β-galactosidase, and glucose isomerase, respectively). Calibration graphs were rectilinear from 0.2 µM to 1 mM glucose, -sucrose and -maltose (limit of detection 0.1 µM) and 3 µM to 1 mM lactose and -fructose (limit of detection 1 µM). Analysis time was ~2 min.
Pharmaceutical Chemiluminescence Controlled pore glass Immobilized enzyme

"Spectrophotometric Flow Injection Determination Of Sucrose And Total Reducing Sugar In Sugar-cane Juice And Molasses"
Anal. Chim. Acta 1988 Volume 214, Issue 1-2 Pages 247-257
I. L. Mattos, E. A. G. Zagatto and A. O. Jacintho

Abstract: Samples of sugar-cane juice or molasses were injected into the carrier stream (H2O at 1 mL min-1) which was then mixed with 1 M HCl in a 250-cm PTFE coil for hydrolysis of sucrose. This solution then merged with 3 M NaOH - 1%K3Fe(CN)6 reagent solution in a 580-cm PTFE coil, the solution was debubbled and 0.1% 1,10-phenanthroline - 0.5 M acetic acid - 0.1 M citric acid - 30 mg L-1 of Fe(III) was added. The solution passed through a 200-cm polyethylene coil and the absorbance was measured at 512 nm. The sampling rate for determination of sucrose and total reducing sugar was 40 h-1. The sampling rate was almost doubled when only reducing sugar was determined and a simplified flow system (illustrated) could be used. The coefficient of variation for determination of sucrose and total reducing sugar in sugar-cane juice were 0.47 and 0.38, respectively; those for molasses were 0.54 and 0.44%. Results generally agreed well with those from classical methods.
Vegetable Juice Spectrophotometry Debubbler Merging zones Method comparison

"Hybrid Biosensor For The Determination Of Sucrose"
Anal. Chim. Acta 1991 Volume 247, Issue 1 Pages 83-87
A. Barlíková, J. Svorc and S. Miertus

Abstract: Yeast cell walls of Saccharomyces cerevisiae as a source of invertase together with glucose oxidase were used for the construction of a hybrid sucrose sensor. The biocatalytic layer was prepared by co-immobilizing yeast cell walls and glucose oxidase on a nylon network via glutardialdehyde and was fixed to the Clark oxygen electrode. The mutarotation of α- to β-glucose was accelerated by phosphate ions instead of mutarotase. The influence of pH, temperature and optimum composition of the biocatalytic layer was tested. Test samples and real samples were examined and the results were compared with those obtained by means of a glucose electrode. The sensor is stable for more than 2 months and the linear range extends to 1.3 x 10^-3 mol l-1.
Electrode Sensor Reactor

"Flow Injection Systems For The Determination Of Oxidoreductase Substrates: Applications In Food Quality Control And Process Monitoring"
Anal. Chim. Acta 1991 Volume 249, Issue 1 Pages 137-143
I. Ogbomo, R. Kittsteiner-Eberle, U. Englbrecht, U. Prinzing, J. Danzer and H. -L. Schmidt

Abstract: Flow injection fluorimetric analysis systems incorporating enzyme reactors are described. The first system, for determination of glucose (I) and ethanol in baker's yeast fermentations, contains a microfiltration module (described) for exclusion of micro-organisms, a glucose dehydrogenase reactor and an alcohol dehydrogenase reactor. The second system, for determination of I and sucrose in fruit juice and lemonade, contains three β-fructosidase reactors in series, a mutarotase reactor and a glucose dehydrogenase reactor. The third system, for determination of I and pullulan contains an α-dextrin endo-1,6-α-glucosidase reactor, an amylglycosidase reactor and a glucose dehydrogenase reactor. All systems require NAD+ in the carrier stream.
Fruit Fluorescence Enzyme Filtration Reactor Process monitoring

"Sequential Determination Of Glucose, Fructose And Sucrose By Flow Injection Analysis With Immobilized Enzyme Reactors And Spectrophotometric Detection"
Anal. Chim. Acta 1992 Volume 261, Issue 1-2 Pages 137-143
Cándido García de María and Alan Townsend*

Abstract: The flow system described and illustrated incorporates two injection valves, one preceding and one after a reactor (R3) containing β-fructofuranosidase, reactors containing hexokinase (R2), phosphoglucose isomerase - glucose-6-phosphate dehydrogenase (R4) and glucose-6-phosphate dehydrogenase alone (R1 and R5), and a flow cell monitored by a spectrophotometer. All enzymes are immobilized on controlled porosity glass (Masoom and Townshend, Ibid., 1985, 171, 185). A solution of ATP, NADP+ and Mg2+ in phosphate buffer (pH 8.0) is used as carrier. Three sequential injections are carried out for each sample; the flow passes through R2, R5 and then R1 for glucose; R3, R2, R5 and R1 for glucose plus sucrose; and R2, R4 and R1 for glucose plus fructose. The method shows good selectivity, and covers the range 0.01 to 1 mM. A flow injection spectrophotometric procedure is described for the sequential determination of glucose, fructose, and sucrose in a sample solution by using single and dual immobilized enzyme reactors in a single manifold. Concentrations in the range 10^-5-10-3 M are determined with anal. rates of 30-40 determinations h-1. A relative standard deviation of 0.8% was obtained for glucose and sucrose and of 1.1% for fructose (10 samples of 4 x 10^-4 M in all cases).
Spectrophotometry Immobilized enzyme Controlled pore glass Differential detection

"Simultaneous Determination Of Sucrose And Reducing Sugars Using Indirect Flow Injection Biamperometry"
Anal. Chim. Acta 1993 Volume 271, Issue 2 Pages 239-246
Jacek Michawski and Anatol Koj, Marek Trojanowicz* and Bogdan Szostek, Elias A. G. Zagatto

Abstract: The cited method used a hexacyanoferrate(III) - hexacyanoferrate(II) indicating system. The flow injection system consisted of PTFE heating coils (0.7 mm), immersed in boiling water, and two injection valves. Two 150 µL portions of the sample were injected via the two valves with a time delay of 30 s. The first sample reacted with hexacyanoferrate(III); the second sample was hydrolyzed and then reacted with hexacyanoferrate(III). The resulting hexacyanoferrate(II) was detected. Changes in concentration. were monitored amperometrically with Pt wire electrodes polarized at 200 mV. Sucrose and glucose were determined with a sampling rate of 40 h-1. The method was applied to samples from white beet juice and syrups from stages of sugar production.
Food Juice Biamperometry Electrode Dual detection Indirect Heated reaction

"Multi-site Detection In Flow Analysis. 3. Periodate Tubular Electrode With Low Inner Volume As A Relocatable Detector"
Anal. Chim. Acta 1994 Volume 285, Issue 3 Pages 293-299
José A. Gomez Neto, Ana Rita A. Nogueira, H. Bergamin Filho and Elias A. G. Zagatto*, José L. F. Costa Lima and Conceição B. S. M. Montenegro

Abstract: The membrane sensor comprised 30% PVC, 63% o-nitrophenyl octyl ether and 7% tetra-octylammonium periodate, with an inner volume of 45 µL. Its position could be changed with use of an electronic commutator, to which were attached the working and Ag/AgCl reference electrodes. Two configurations are described. In one, used for the determination of glycerol, the sensor could be moved to either of two identical channels. Glycerol solution (30 µL) were injected into a reagent carrier of 3 mM NaIO4, 0.5 M Na2SO4, 0.1 M acetic acid and 0.1 M sodium acetate (0.5 ml/min). The carrier passed through a coiled reactor (1 m x 0.7 mm i.d.) to the sensor, where unconsumed periodate was measured. Calibration graphs were linear for 0.05-0.2% glycerol. The commutator was switched to the parallel channel a few s after the peak maximum was reached, thus the sampling rate of 100/h was not affected by the wash-out time. For sucrose, the sensor was moved to a position downstream in the same channel. Calibration graphs were linear for 0.1-0.5% sucrose. Glycerol was determined in soaps and detergents, and sucrose was determined in sugar cane juice and syrups.
Commercial product Juice Syrup Electrode Sensor Apparatus Detector

"Application Of Enzyme Field-effect Transistor Sensor Arrays As Detectors In A Flow Injection System For Simultaneous Monitoring Of Medium Components. 1. Preparation And Calibration"
Anal. Chim. Acta 1994 Volume 296, Issue 3 Pages 263-269
T. Kullick, M. Beyer, J. Henning, T. Lerch, R. Quack, A. Zeitz, B. Hitzmann, T. Scheper and K. Schügerl*

Abstract: Enzymes were co-immobilized on the pH-sensitive gates of an 8-channel array of field-effect transistors (FETs). Glucose was determined with a glucose dehydrogenase (GDH) FET, maltose with a co-immobilized maltase (MAL)/GDH FET, sucrose with a co-immobilized invertase (INV)/GDH FET, lactose with a β-galactosidase/galactose dehydrogenase (β-GAL/GALDH)-fusion protein FET and ethanol with a co-immobilized alcohol dehydrogenase/aldehyde dehydrogenase (ADH/ALDH) FET. These EnFETs were integrated into FIA systems, and were calibrated and characterized with respect to pH, buffer capacity, stirrer speed, NAD concentration and cross sensitivity. Because the signals of the EnFETs were sensitive to pH and buffer capacity, the pH was monitored with a pH-FET in the array, and the buffer capacity and substrate concentration were calculated from the shape of the signal of the FIA system. The EnFETs were stored at 4°C for many months without activity loss. They have satisfactory activity for performing a large number of analyzes. However, glucose reversibly inhibited the sucrose signal and added to the maltose signal. Hence the sucrose monitor can only be used when glucose is practically absent, and the maltose sensor requires simultaneous determination of glucose.
Fermentation broth Field effect transistor Sensor Simultaneous analysis pH Buffer Optimization

"Application Of Enzyme Field-effect Transistor Sensor Arrays As Detectors In A Flow Injection System For Simultaneous Monitoring Of Medium Components. 2. Monitoring Of Cultivation Processes"
Anal. Chim. Acta 1995 Volume 300, Issue 1-3 Pages 25-31
T. Kullicka, U. Bocka, J. Schuberta, T. Scheperb and K. Schügerla,*

Abstract: A biosensor array consisting of a single pH-FET and seven enzyme FET channels integrated into a single flow injection system was used for the online monitoring of pH, glucose, maltose, sucrose, lactose, ethanol and urea during the cultivation of Escherichia coli and Saccharomyces cerevisiae in synthetic media. The components were monitored using glucose dehydrogenase, maltase-glucose dehydrogenase, invertase-glucose dehydrogenase, β-galactosidase-galactose dehydrogenase, alcohol dehydrogenase-aldehyde dehydrogenase and urease (co)immobilized on the pH sensitive gates of the FET. The array was integrated in a commercial FIA system. The results agreed with those obtained by offline concentration measurements.
Fermentation broth Field effect transistor Field effect transistor Sensor Method comparison

"Development Of Biosensors Based On An Electrolyte Isolator Semiconductor (EIS)-capacitor Structure And Their Application For Process Monitoring. 1"
Anal. Chim. Acta 1995 Volume 317, Issue 1-3 Pages 259-264
C. Menzel, T. Lerch, T. Scheper and K. Schügerl*

Abstract: Simple non-structured fluoride sensitive electrolyte isolator semiconductor (EIS) chips consisting of Si/SiO-2/Si-3N-4/LaF-3 layers were used as transducers for measuring the H-2O-2 concentration by peroxidase (POD) and combined this H-2O-2 sensor with various oxidases, which form H-2O-2 in stoichiometric amount. The concentrations of the following analytes were measured with this system: glucose with glucose oxidase (GOD) and POD, maltose with amyloglucosidase (AGLU), mutarotase (MUT), GOD and POD, amylose with β-amylase (Amyl), AGLU, MUT, GOD and POD, sucrose with invertase (INV), MUT, GOD and POD, ethanol with alcohol oxidase and POD, sulfite with sulfiteoxidase (SOD) and POD, xanthine with xanthineoxidase (XAD) and POD, phosphate with nucleoside-phosphorylase (NP), XOD and POD. The required enzymes were coimmobilized on the surface of the transducer (pF-EIS-CAP), mounted into a flow cell, equipped with a reference electrode, integrated into a flow injection analyzer. system and operated with the CAFCA (Computer Assisted Flow Control and Analysis) automation programme. (11 references)
Sensor Sensor Immobilized enzyme Process control Process monitoring

"Copper Dispersed Into Polyaniline Films As An Amperometric Sensor In Alkaline Solutions Of Amino-acids And Polyhydric Compounds"
Anal. Chim. Acta 1996 Volume 335, Issue 3 Pages 217-225
Innocenzo G. Casellaa,*, Tommaso R. I. Cataldia, Antonio Guerrieria and Elio Desimonib

Abstract: A vitreous C electrode (0.125 cm2 area) was coated with a polyaniline (PANI) film by galvanostatic polymerization from a solution containing 85 mM aniline in 0.1 M H2SO4 by cycling the potential between -0.1 and 1.1 V (vs. SCE) for five cycles at 50 mV/s. The coated electrode was immersed in 50 mM CuCl2 in 0.1 M H2SO4 for 5 min and then a potential of -0.3 V was applied for 3 min. The Cu-PANI sensor was evaluated for determining amino-acids and polyhydric compounds. Molar response factors for carbohydrates and amino-acids were measured using a FIA system and 0.1 M NaOH as the carrier stream (1 ml/min). A potential of 0.55 V vs. Ag/AgCl was applied to the sensor. A Carbopac PA 1 anion-exchange column (25 cm x 4 mm i.d.) was coupled to the FIA system to analyze carbohydrates and amino-acids. Xylitol, sorbitol, glucosamine, glucose, lactose and sucrose were separated using 0.15 M NaOH as the mobile phase (0.6 ml/min) and the detection limits were 2-6 pmol. Alanine, glycine, lysine methionine and glutamine were separated with 0.1 M NaOH as the mobile phase (0.5 ml/min) and the detection limits were 5-15 pmol. The linear dynamic range was three-four orders of magnitude above the detection limits. The electrode was stable for >=3 h in flowing streams.
Electrode Electrode Amperometry Electrode Ion exchange Sensor Linear dynamic range

"Determination Of Sucrose Using Sucrose Phosphorylase In A Flow Injection System"
Anal. Chim. Acta 1997 Volume 337, Issue 1 Pages 107-111
Mamie Kogure, Hisakazu Mori*, Hisano Ariki, Chika Kojima and Haruhiko Yamamoto

Abstract: The fluorimetric determination of sucrose was accomplished using a single-channel flow manifold incorporating an enzyme reactor (8 cm x 2 mm i.d.) prepared by immobilizing 5 iu sucrose phosphorylase/11 iu phosphoglucomutase/4 iu glucose 6-phosphate dehydrogenase on to 0.4 g aminopropyl glass beads (200-400 mesh, 500 angstrom pore size). The determination was carried out by injecting a sample (50 µL) into a carrier stream at a flow rate of 0.4 ml/min containing 30 mM KH2PO4/3 mM MgCl2/1 mM NADP+/0.05 mM glucose 1,6-diphosphate in 200 mM PIPES buffer of pH 7.5. The stream was propelled through the enzyme reactor at 30°C and into the detection cell (12 µL volume); detection was at 460 nm (excitation at 340 nm). The calibration graph was linear from 0.1-200 µM sucrose; the detection limit was 0.1 µM. The method was applied to the determination of sucrose in soft drinks. The samples were diluted 1000-fold prior to analysis and the phosphate concentration in the carrier stream was increased to 100 mM to reduce the inhibiting effects of fructose. The detection limit was 0.2 µM sucrose. RSD was 1.06% (n = 5) for 50 µM-sucrose.
Soft drink Fluorescence Immobilized enzyme Glass beads Heated reaction

"Continuous-flow Determination Of Reducing Sugars And Sucrose In Natural And Industrial Products With Periodate Oxidation And A Periodate-sensitive Flow-through Electrode"
Analyst 1982 Volume 107, Issue 1281 Pages 1471-1478
E. P. Diamandis and T. P. Hadjiioannou

Abstract: A continuous-flow method for the determination of mixtures of reducing sugars and sucrose in natural and industrial products is described. The sample, before and after hydrolysis of sucrose, reacts with an excess of periodate in a flow system and the decrease in periodate activity is monitored with a periodate-sensitive flow-through electrode. The recorded peak heights are indirectly linearly related to the concentration of reducing sugars in the range 3-18 mM. The sucrose concentration is calculated by difference. The analysis is completely automated, requires no sample pre-treatment except for the hydrolysis of sucrose and samples can be analyzed at a rate of 30 per hour with a relative error and a relative standard deviation of 1-3%. Comparison with Fehling's method for various natural and industrial products gave satisfactory results.
Agricultural Industrial Electrode Electrode

"Chemically Immobilized Tri-enzyme Electrode For The Determination Of Sucrose Using Flow Injection Analysis"
Analyst 1988 Volume 113, Issue 1 Pages 81-85
Junainah Abdul Hamid, G. J. Moody and J. D. R. Thomas

Abstract: Invertase, mutarotase and glucose oxidase were co-immobilized on a nylon mesh which was then placed over a platinum electrode. This electrode was used in the amperometric mode in a three-electrode Stelte cell adapted for flow injection analysis for monitoring the hydrogen peroxide enzymolysis product. The optimum enzyme composition (I.U. ratios) for immobilization on the nylon net of invertase-mutarotase-glucose oxidase was found to be in the ratio 2000 : 1000 : 200. The system exhibited good linearity in the range 10^-6-10-3 M sucrose, short response times (10-20 s), a long lifetime (6% reduction in signals after >14 h continuous flow of fresh 1 mM sucrose) and good storage stability (38 d, stored in 0.1 M pH 7 phosphate buffer at 4°C). The nylon mesh enzyme electrode response and excellent resistance towards radiation, i.e., sterilisation (2.1 Mrad of 60Co -source), indicated its potential use for on-line monitoring of sucrose in clinical and food-processed samples.
Amperometry Clinical analysis Electrode Immobilized enzyme Interferences Optimization

"Repetitive Determinations Of Amylase, Maltose, Sucrose And Lactose By Sample Injection In Closed Flow-through Systems"
Anal. Chem. 1978 Volume 50, Issue 12 Pages 1665-1670
D. P. Nikolelis and Horacio A. Mottola

Abstract: Conditions have been developed for the repetitive determination of amylase and some disaccharides by coupling enzyme-catalyzed reactions yielding glucose as a product with the glucose oxidase catalyzed oxidation of this sugar. Determinations have been performed by sample injection into a continuously circulated reagent mixture and monitoring of oxygen depletion with a three-electrode amperometric system. Maltose, sucrose, and lactose in the range of 50 to 500 mg/100 mL, 10 to 250 mg/100 mL, and 25 to 250 mg/100 mL, respectively, and amylase in the range of 50 to 500 units/100 mL have been determined with relative errors and relative standard deviations (population) of about 2%. The maximum determination rate is 240 injections/h for maltose, 700 injections/h for sucrose and lactose; and 120 injections/h for amylase at room temperature. Applications to real samples (a variety of food products, human blood serum, and serum calibration references and controls) are reported.
Food Serum Human Clinical analysis Electrode Closed loop

"Chemiluminescence Flow Injection Analysis Determination Of Sucrose Using Enzymatic Conversion And A Microporous Membrane Flow Cell"
Anal. Chem. 1986 Volume 58, Issue 1 Pages 116-119
Cathy A. Koerner and Timothy A. Nieman

Abstract: Sucrose is initially converted into glucose by β-fructofuranosidase and aldose-1-epimerase (immobilized on controlled-porosity glass) in 0.1 M phthalate buffer (pH 5.65). The solution is then mixed with a stream containing luminol, haemin and horse-radish peroxidase in 0.1 M Tris (pH 10.5) and passes finally into a flow cell where it mixes with glucose oxidase in 0.1 M acetate buffer (pH 5). The H2O2 formed reacts with the luminol to produce chemiluminescence. The working range is 5 µM to 1 mM and analysis time is 2 min. For detection of sucrose in food products, a separate determination of glucose or a catalytic destruction of glucose in the sample is necessary to distinguish glucose originally present from that formed from sucrose.
Soft drink Breakfast Mix Chemiluminescence Immobilized enzyme Microporous membrane Flowcell

"Pulsed Amperometric Detection Of Carbohydrates At Gold Electrodes With A Two-step Potential Waveform"
Anal. Chem. 1987 Volume 59, Issue 1 Pages 150-154
Glen G. Neuburger and Dennis C. Johnson

Abstract: Glucose, glucitol and sucrose in 0.2 M NaOH were determined by flow injection analysis with pulsed amperometric detection at a gold electrode. Where necessary, O was removed by purging with Ar. Potentials were measured against the SCE and a two-step potential waveform was obtained. Detection limits were ~1 nmol in a 50 µL sample. The method can also be used following separation by HPLC.
Amperometry Electrode

"Simultaneous Determination Of Glucose, Fructose, And Sucrose In Mixtures By Amperometric Flow Injection Analysis With Immobilized Enzyme Reactors"
Anal. Chem. 1988 Volume 60, Issue 2 Pages 147-151
Kiyoshi Matsumoto, Hideaki Kamikado, Hiroaki Matsubara, and Yutaka Osajima

Abstract: The flow injection system (described) incorporated three immobilized-enzyme reactors (prep. described), one each for glucose(I), fructose(II) and sucrose(III), in parallel, a I-eliminating enzyme reactor (placed in series with the III reactor), and a multi-channel flow-through amperometric detector. Sample solution was injected into the system by an injection valve and into each of the three carrier streams by valve switching. For I determination, the carrier stream was 0.1 M phosphate buffer (pH 6.0), the reactor contained immobilized I oxidase, and the H2O2 produced was determined at +0.65 V vs. Ag - AgCl. For II determination, the carrier stream was McIlvaine buffer (pH 5.0) containing 6 mM K3Fe(CN)6 and 0.1% of Triton X-100, the reactor contained immobilized II 5-dehydrogenase, and the Fe(CN)64- formed was determined at +0.385 V vs. Ag - AgCl. For III determination, the carrier stream was 0.1 M phosphate buffer (pH 7.0), the reactor contained immobilized β-fructofuranosidase, aldose 1-epimerase and I oxidase, and the H2O2 formed was determined as before. The coefficient of variation (n = 10) for 1 mM I, -II and -III were 1.8, 1.8 and 1.6%, respectively. Calibration graphs (response vs. concentration.) were rectilinear from 0.02 to 1 mM for I, II and III. The method was applied to food samples.
Food Soft drink Fruit Amperometry Immobilized enzyme Multicomponent Triton X Valve Surfactant

"Catalytic Oxidation And Flow Detection Of Carbohydrates At Ruthenium Dioxide-modified Electrodes"
Anal. Chem. 1990 Volume 62, Issue 14 Pages 1413-1416
Joseph Wang and Ziad Taha

Abstract: A RuO2-modified carbon paste electrode (prep. described) was used in the constant-potential cyclic voltammetric flow detection of carbohydrates. Samples in 1 M NaOH were analyzed at the electrode (+0.4 V; 20 mV s-1) with Ag - AgCl and Pt-wire reference and auxillary electrodes, respectively. The effects of variables, viz, pH, flow rate, operating potential and surface 'loading', were studied. The method was applied in the detection of deoxyribose, fructose, galactose, gluconic acid, glucose, glycerol, lactose, maltose, ribose and sucrose (electrode responses are tabulated). Working ranges were from 0.1 to 10 mM glucose and 0.1 to 1 mM ribose, fructose and galactose. The detection limits were pmol-levels and the coefficient of variation (n = 72) was 1.2%. The electrode was stable for >48 h with a signal loss of 10% over this period.
Electrode Electrode Voltammetry Detection limit Optimization Catalysis

"Liquid-interfaced Oscillating Glow Discharge Detector For A Flowing Liquid System"
Anal. Chem. 1995 Volume 67, Issue 5 Pages 878-884
Christopher J. Herring and Edward H. Piepmeier

Abstract: A new liquid-interfaced oscillating glow discharge detector having a frequency and current response to femtomole and picomole quantities respectively of potassium nitrate and sucrose injected into an aqueous flowing eluent is presented. The glow discharge is formed in an argon atmosphere at ambient pressure between a platinum anode and a cathode consisting of an aqueous conducting solution. A detailed description of the appearance of the liquid-interfaced glow discharge at various electrode distances and the occurrence of high-frequency oscillations is given. Copyright 1995, American Chemical Society.
Spectrophotometry Interface

"Determination Of Sucrose By Flow Injection Analysis With Fourier Transform Infrared Detection"
Microchim. Acta 1995 Volume 119, Issue 1-2 Pages 73-79
Bernhard Lendl and Robert Kellner

Abstract: The cited analysis was based on the invertase-catalyzed cleavage of sucrose (I) to α-D-glucose and β-D-fructose in a reactor (3 cm x 3 mm i.d.) packed with invertase immobilized on aminopropylated controlled pore glass using glutaraldehyde (details given). A manifold (diagram given) incorporating two internally-coupled injection valves enabled FTIR spectra of the unreacted and reacted sample to be obtained. Sample (75 µL) was injected into 0.2 M acetate buffer of pH 4.2 (2.1 ml/min) and FTIR spectra were recorded continuously at 8 cm-1 resolution with use of Brucker OPUS GC-IR software and the difference spectrum of the unreacted and reacted sample was obtained. The difference in absorption at 998 and 1038 cm-1 was used to determine I. The calibration graph was linear for 10^-100 mM I. No detection limit or RSD is given. Sample throughput was 45/h. The method was applied to soft drinks.
Soft drink Spectrophotometry Immobilized enzyme Controlled pore glass

"Simultaneous Determination Of Glucose And Sucrose By A Dual-working-electrode Multi-enzyme Sensor Flow Injection System"
Electroanalysis 1994 Volume 6, Issue 5-6 Pages 361-367
Xianen Zhang, Garry A. Rechnitz

Abstract: Two working electrodes, each consisting of carbon paste covered with a cross-linked enzyme membrane, were mounted in a flow injection flow cell (diagram presented). The membrane of the upstream electrode was prepared from glucose oxidase and that of the downstream electrode from glucose oxidase with an outer layer of β-fructofuranosidase/aldose 1-epimerase. A layer of cross-linked catalase was formed on the wall of the flow cell between the two electrode membranes. A 50 µL sample was injected into a carrier stream of phosphate buffer solution (pH 6.8); both electrodes were maintained at 0.9 V vs. Ag/AgCl, so that glucose was detected at the upstream and glucose plus sucrose at the downstream electrode. The method of calculating the two concentrations is described. When the method was applied to Cola beverages, the relative errors for glucose and sucrose were ~3 and ~4%, respectively.
Soft drink Amperometry Electrode Electrode Electrode Sensor

"Applications Of Immobilized Enzymes In Flow Injection Analysis"
Anal. Proc. 1985 Volume 22, Issue 1 Pages 6-8
Stephen, M. Masoom, Alan Townshend

Abstract: Samples of blood serum and soft drinks were injected into a stream of phosphate buffer solution (pH 7) which then passed through a glass column (2.5 cm x 2.5 mm) containing glucose oxidase (immobilized on controlled-pore glass) to a flow-through amperometric detector for the determination of H2O2. Sucrose in soft drinks was determined by use of immobilized β-D-fructofuranosidase, aldose 1-epimerase and glucose oxidase. The limit of detection was 1 µM-H2O2.
Beverage Blood Serum Amperometry Immobilized enzyme Controlled pore glass

"A New Version Of An In-situ Sampling System For Bioprocess Analysis"
Acta Biotechnol. 1996 Volume 16, Issue 2-3 Pages 185-192
J.-M. Hilmer, Th. Scheper

Abstract: In this article, attention is focused on an online sampling device for bioreactors and its characterization regarding sterility and response time. The integration of this device into analytical systems for biotechnology (FIA with biosensors) resulted in online analysis systems for bioprocess control. The new ESIP version of an in situ sampling system for bioprocess analysis produced by the EPPENDORF-NETHELER-HINZ GmbH, Germany, had a short response time of 8 min (99%), which was determined by means of conductivity measurement after an increase of the medium conductivity due to the gradual addition of KCl. Effectiveness and reliability of the module were tested by bubble point measurement resulting in a bubble point pressure of 2.1 bars. The sampling probe was tested successfully for use in a broad variety of microorganisms and cultivations.
Fermentation broth Sensor Biotechnology Process control Review

"Hyphenation Of Sequential- And Flow Injection Analysis With FTIR-spectroscopy For Chemical Analysis In Aqueous Solutions"
AIP Conf. Proc. 1998 Volume 430, Issue 1 Pages 403-406
B. Lendl, R. Schindler, and R. Kellner

Abstract: A survey of the principles of sequential (SIA)- and flow injection analysis (FIA) systems with FTIR spectroscopic detection is presented to introduce these hyphenations as powerful techniques for performing chemical anal. in aqueous solution The strength of FIA/SIA-FTIR systems lies in the possibility to perform highly reproducible and automated sample manipulations such as sample clean-up and/or chemical reactions prior to spectrum acquisition. It is shown that the hyphenation of FIA/SIA systems with an FTIR spectrometer enhances the problem solving capabilities of the FTIR spectrometer as also parameters which can not be measured directly (e.g. enzyme activities) can be determined On the other hand application of FTIR spectroscopic detection in FIA or SIA is also of advantage as it allows to shorten conventional anal. procedures (e.g. sucrose or phosphate anal.) or to establish and apply a multivariate calibration model for simultaneous determinations (e.g. glucose, fructose and sucrose anal.). In addition to these examples two recent instrumental developments in miniaturized FIA/SIA-FTIR systems, a µ-Flow through cell based on IR fiber optics and a micromachined SI-enzyme reactor are presented in this paper.
Spectrophotometry Interface Sequential injection Multivariate calibration Optical fiber

"Hyphenation Of Sequential- And Flow Injection Analysis With FTIR-spectroscopy For Chemical Analysis In Aqueous Solutions"
AIP Conf. Proc. 1998 Volume 430, Issue 1 Pages 403-406
B. Lendl, R. Schindler, and R. Kellner

Abstract: A survey of the principles of sequential (SIA)- and flow injection analysis (FIA) systems with FTIR spectroscopic detection is presented to introduce these hyphenations as powerful techniques for performing chemical anal. in aqueous solution The strength of FIA/SIA-FTIR systems lies in the possibility to perform highly reproducible and automated sample manipulations such as sample clean-up and/or chemical reactions prior to spectrum acquisition. It is shown that the hyphenation of FIA/SIA systems with an FTIR spectrometer enhances the problem solving capabilities of the FTIR spectrometer as also parameters which can not be measured directly (e.g. enzyme activities) can be determined On the other hand application of FTIR spectroscopic detection in FIA or SIA is also of advantage as it allows to shorten conventional anal. procedures (e.g. sucrose or phosphate anal.) or to establish and apply a multivariate calibration model for simultaneous determinations (e.g. glucose, fructose and sucrose anal.). In addition to these examples two recent instrumental developments in miniaturized FIA/SIA-FTIR systems, a µ-Flow through cell based on IR fiber optics and a micromachined SI-enzyme reactor are presented in this paper.
Spectrophotometry Interface Sequential injection Multivariate calibration Optical fiber

"Supersensitive Analytical Detection By Voltammetry"
Am. Lab. 1998 Volume 30, Issue 1 Pages 21-25
Sturrock, P.E.;Barringer, G.E.

Abstract: This paper describes an improved instrument (DP90, Groton Technol., Acton, MA), the design of which was based on experience with the DP82. The DP90 can be used in a traditional manner in unstirred bulk solution or in flow streams as detectors for HPLC or FIA. It operates with a commercial PC as a host computer and features a real-time display of operating parameters and acquired data, the unique ability to change parameters repeatedly during an experiment, the capacity to store all data and parameter values, and the capability of compensating background currents on a point-by-point basis during voltammetric sweeps to allow increased gain in the instrument. (SFS)
Voltammetry Sensitivity

"Theoretical Consideration On Polarized Photometric Detection"
Biomed. Chromatogr. 1994 Volume 8, Issue 3 Pages 130-133
Kazuichi Hayakawa, Atsushi Yamamoto, Akinobu Matsunaga, Eiichi Mizukami, Masayuki Nishimura, Motoichi Miyazaki

Abstract: Previously, we developed a novel detector, a polarized photometric detector for optically active compounds, where two polarizers are set on either side of the UV-visible absorbance detector flow cell. In this paper, we present theoretical considerations on this method with several equations. If the inclined angle of the second polarizer is defined as α (left when α > 0°), the following equation is obtained: ΔAbs = 2 log e.tan α.beta, where ΔAbs is the change in absorbance and beta (magnitude of beta << α) is the angle of rotation of light passing through the cell containing the optically active compound. This equation suggests that ΔAbs is proportional to the analyte concentration and that the dextro- and laevorotatory compounds are detected as positive and negative absorbance changes, respectively. The theoretical maximum signal to noise (S/N) ratio is obtained when α = 45° (= 0.8 rad) and the smallest detection limit is 5 x 10^-5° (S/N = 3) when the path length of the flow cell is 10 mm. This detector was successfully used in flow injection analysis (FIA) as well as HPLC for optically active compounds.
Pharmaceutical Spectrophotometry Polarimetry HPLC Theory

"Biosensing Of Glucose, Sucrose, And Lactate In Beverages With An Automated Multi-channel Flow Analyzer"
Biosci. Biotechnol. Biochem. 1995 Volume 59, Issue 5 Pages 813-816
Chen, R.L.C.;Matsumoto, K.

Abstract: A multi-channel continuous-flow analyzer equipped with biosensing devices was developed for multicomponent measurement and its use in automating routine analysis was evaluated. Biosensing was achieved by the aid of an immobilized enzyme reactor installed in the channel, and the channel switching process for the sensing of a different compound was made by using a column-switching rotary valve. Another rotary valve was used for auto-sampling. Both of the two rotary valves were interfaced to a system controller and work conjugatively in a programmed manner. Signal subtraction between different channels was found to be more precise compared with the multi-channel flow injection analysis method, which is of merit for an analysis utilizing enzyme relay reaction (as for sucrose analysis) or for background signal subtraction. Glucose, lactate, and sucrose content in real samples were measured automatically with high reproducibility, and the results agree well with the kit method.
Beverage Immobilized enzyme Valve Multichannel

"Evaluation Of Polypyrrole - Glucose Oxidase Electrodes In Flow Injection Systems For Sucrose Determination"
Biosens. Bioelectron. 1991 Volume 6, Issue 3 Pages 263-273
Wolfgang Schuhmann and Ruth Kittsteiner-Eberle

Abstract: Thin films of polypyrrole were deposited on Pt or vitreous-carbon disc electrodes (1 or 3 mm diameter) by electropolymerization of the monomer. The polymer film was nitrated by treatment with acetic anhydride and Cu(NO3)2 and the nitro-groups were reduced electrochemically to amino-groups. The glucose oxidase was immobilized on the electrode surface via the formation of amides or secondary amines (full details given). The electrodes were operated at a working potential of +600 mV (vs. the SCE), and in the flow injection mode, with 0.5 M NaCl - 0.1 M phosphate (pH 7.4) as supporting electrolyte, responses were rapid (~10 s) and rectilinear for 0.05 to 10 mM glucose. The response fell to 60 to 80% of its initial value after 2 days, but then remained fairly constant for a further 20 to 23 days. Attempts to determine simultaneously glucose and sucrose in the same solution, by use of two electrochemical sensors, in combination with immobilized β-fructofuranosidase (I) and aldose 1-epimerase (II), were unsuccessful owing to inhibition of glucose oxidase by the relatively high concentration. of sucrose. However simultaneous determination of glucose and sucrose was possible in a flow injection system containing enzyme columns with glucose dehydrogenase, I and II immobilized on controlled-pore glass, with fluorimetric determination of enzymatically generated NADH.
Electrochemical analysis Electrode Electrode Electrode Electrode Fluorescence Column Controlled pore glass Immobilized enzyme

"An Amperometric Flow Injection Analysis Enzyme Sensor For Sucrose Using A Tetracyanoquinodimethane Modified Graphite-paste Electrode"
Biosens. Bioelectron. 1996 Volume 11, Issue 8 Pages 719-723
J. L. Lima Filho, P. C. Pandey*, H. H. Weetall

Abstract: FIA systems for the determination of sucrose are described in which (i) invertase and µarotase were immobilized on to controlled pore glass beads in an enzyme reactor and the C-paste detection electrode was modified with glucose oxidase (I) and tetracyanoquinodimethane (II) as electron transfer mediator or (ii) invertase was incorporated into the C-paste electrode together with I and II (an enzyme reactor was not used). Both systems were operated with 0.1 M phosphate buffer of pH 7 as the carrier stream (30 ml/h) and a 250 µL sample loop. The amperometric response was measured at 200 mV vs. Ag/AgCl. The detection range was 0.025-200 mM for i and up to 2 M for ii.
Amperometry Electrode Electrode Sensor Controlled pore glass

"Rapid Determination Of Glucose And Sucrose By An Amperometric Glucose-sensing Electrode Combined With An Invertase/mutarotase-attached Measuring Cell"
Biosens. Bioelectron. 1997 Volume 12, Issue 9-10 Pages 1013-1020
Fumio Mizutania,* and Soichi Yabukia

Abstract: Glucose and sucrose are simultaneously determined by the use of an enzyme sensor system consisting of a glucose-sensing electrode based on a lipid-modified glucose oxidase and a measuring cell that contains an invertase/mutarotase-coimmobilized layer. From the current response of the enzyme electrode after the addition of a glucose/sucrose mixture, the concentrations of the two kinds of sugars can be separately determined: the concentration of glucose (0 . 2 µM-3 mM) is determined from the steady-state current increase obtained from 2 to 6s after the addition of the mixture, and that of sucrose (10 µM-6 mM), from the rate of current increase from 8 to 20 s after the addition. The relative standard deviations are 1 . 7% for glucose and 3 . 1% for sucrose (n = 10). The system can be applied to the rapid determination of glucose and sucrose in food samples.
Food Amperometry Sensor Electrode Nafion membrane Simultaneous analysis

"Flow Injection Analysis System For The Supervision Of Industrial Chromatographic Downstream Processing In Biotechnology"
Biosens. Bioelectron. 1998 Volume 13, Issue 12 Pages 1251-1255
P. Sosnitzaa, F. Irtela, R. Ulbera, M. Busseb, R. Faurieb, L. Fischerc and T. Schepera,*

Abstract: Sugar beet molasses is a natural resource for various products used in daily life, ranging from sucrose to amino acids for pharmaceutical industry. The separation of molasses into these high value components is performed on a large scale by ion exchange/exclusion chromatography A biosensor system was set up for the 'in time' anal. of serine and sucrose during molasses desugarization. D-Serine was analyzed with the multi-enzyme system D-serine dehydratase/lactic dehydrogenase and photometric detection of the NADH consumed. Sucrose was determined with invertase/mutarotase/glucose oxidase and the oxygen consumed was monitored amperometrically. An anal. could be performed within 2-5 min by directly injecting samples from the chromatography process into the flow injection analysis system. The determination range for the sucrose anal. was 0-2.5 gl-1 and for the anal. of D-serine 0-0.5 gl-1. The standard deviation for the measurement of D-serine was 1.7%.
Food Amperometry Sensor Spectrophotometry HPLC Process monitoring Immobilized enzyme

"Flow Injection Analysis Of Glucose, Fructose And Sucrose Using A Biosensor Constructed With Permeabilized Zymomonas Mobilis And Invertase"
Biotechnol. Prog. 1995 Volume 11, Issue 1 Pages 58-63
Je-Kyun Park, Min-Chol Shin, Seung-Goo Lee, and Hak-Sung Kim

Abstract: Cells of Zymomonas mobilis possessing glucose-fructose oxidoreductase and gluconolactonase were permeabilized with toluene and co-immobilized with invertase within a gelatin membrane. This membrane was coated over a pH electrode, and the resulting biosensor was placed in a flow-through cell to develop a flow injection analysis system for the specific determination of glucose, fructose, and sucrose. Peak height resulting from the production of hydrogen ion was correlated with sugar concentration, and the effect of operating variables on the response characteristics of the FIA system was investigated on the basis of theoretical and experimental analyzes. Under the optimized conditions, the calibration curves for glucose, fructose, and sucrose were linear up to 8, 80, and 60 g/L, respectively. The FIA system was applied to the online monitoring of glucose production in the enzymatic hydrolysis of cellulose, and the glucose concentrations determined using the FIA system coincided well with those determined by the conventional enzymatic method. Copyright 1995, American Chemical Society.
Sensor Electrode Electrode Electrode Immobilized cell Optimization Method comparison

"Simple And Inexpensive Flow Injection Analysis For Determination Of Sucrose Using Invertase And Glucose Oxidase Immobilised On Glass Beads"
Biotechnol. Techniq. 1995 Volume 9, Issue 5 Pages 345-348
V. Leite, I. C. Le&atilde;o, G. F. V. de Vasconcelos, M. C. B. Pimentel, V. L. SIlva, E. H. M. Melo, and J. L. Lima Filho

Abstract: A Flow Injection Analysis (FIA) for sucrose using invertase ( E.C. 3.2.1.26), mutarotase (E.C.5.1.3.3) and glucose oxidase (E.C.1.1.3.4) was developed. The enzymes were immobility on glass beads using glutaraldehyde. The sucrose concentration was related to oxygen saturation. Fall in O-2 concentration, as a result of sucrose oxidation, was detected by a low cost, home-made O-2 electrode. The system was able to measure sucrose from 0.025 to 100 mM with a response time of 6 min using 200 µl of sample, with an apparent K-m of 42 mM of sucrose. The system has been operated satisfactorily for 50 days without loss any initial activity.
Electrode Immobilized enzyme Glass beads Low cost

"Development Of A Sucrose Enzymic Biosensor"
Biotechnol. Techniq. 1998 Volume 12, Issue 4 Pages 305-307
A.M. Salgado, R.O.M. Folly, B. Valdman and F. Valero

Abstract: An enzymatic biosensor for sucrose determination was developed for online and continuous monitoring of sucrose concentration. The sensor was adapted to two different measurement schemes, one continuous and another injection sampling lines. The sensor adapted with the injection sampling line presented a linear measurement range of 5-20 g sucrose/1, good reproducibility, and a high versatility permitting the substitution of the immobilized enzymes when their activity decreased.
Sensor Electrode Apparatus Process monitoring Detector

"Simultaneous Determination Of Components In Food By FIA Including Parallel Configuration Of Enzyme Columns"
Bunseki 1990 Volume 1990, Issue 5 Pages 379-383
Matsumoto, K.

Abstract: A review is presented, with 8 references, of the use of FIA and immobilized-enzyme columns in the determination of components, e.g., glucose, fructose, sucrose, lactic acid and ethanol in food products. Procedures and instrumentation are discussed.
Food Enzyme Column Immobilized enzyme Review

"Flow Injection Determination Of Sugars In Foods By Use Of A Porphinatotitanium(IV) Reagent"
Bunseki Kagaku 1995 Volume 44, Issue 5 Pages 355-362
Yokoi, Y.;Matsubara, C.;Takamura, K.

Abstract: Sample solution (20 µL) was injected into a stream (0.4 ml/min) of 0.05 M phosphate buffer/1 mM MgCl2 (pH 6.6). To determine glucose, the stream passed directly to a column of glucose oxidase, merged with a stream (0.4 ml/min) of 30 µM-oxo-[5,10,15,20-tetra-(4-pyridyl)porphinato]titanium(IV), and then passed through a mixing coil (15 m x 0.5 mm i.d.) maintained at 75°C before absorbance measurement at 450 nm. To determine maltose, lactose or sucrose, the stream after injection of the sample passed first through a column of glucose oxidase/catalase, then through a column of α-glucosidase, β-galactosidase, or β-fructofuranosidase/aldose 1-epimerase, respectively, and then through a column of glucose oxidase before merging with the reagent stream. Calibration graphs for glucose and for maltose, lactose or sucrose were linear over the ranges 0.5-500 µM and 1-1000 µM, respectively, and the respective RSD (n = 10) at 100 µM-glucose, -maltose, -lactose or -sucrose were 0.74, 0.84, 0.49 and 0.75%. The four carbohydrates were determined in milk, soft beverages and wine.
Milk Soft drink Wine Spectrophotometry Immobilized enzyme Heated reaction

"Flow Injection Analytical System For Simultaneous Determination Of Glucose And Sucrose Utilizing Immobilized Enzyme Bioreactors"
Chem. Express 1988 Volume 3, Issue 1 Pages 13-16
Yao, T.;Akasaka, R.;Wasa, T.

Abstract: A flow-injection anal. system with immobilized enzyme bioreactors (invertase-mutarotase-glucose oxidase) is described for the highly selective and highly sensitive simultaneous determination of glucose and sucrose. The simultaneous determination of glucose and sucrose in soft drinks could be performed on 30 samples/h with satisfactory precision (<1% relative standard deviation). (SFS)
Soft drink Immobilized enzyme Multicomponent

"Study On Multiple-enzyme Electrode For Sucrose Determination"
Shengwu Gongcheng Xuebao 1991 Volume 7, Issue 4 Pages 339-344
Hu W, Zhang X, Zhang X, Hu S.

Abstract: Invertase (INV), mutarotase (MUT), glucose oxidase (GOD) and BSA were coimmobilized via glutaraldehyde-bridged covalent bonding, and directly absorbed on the teflon membrane. This membrane was covered with a nylon mesh and placed over an oxygen electrode. An enzyme electrode for flow injection analysis system (EFIA) was adopted. The optimum enzyme composition (IU) for immobilization on the teflon membrane of INV-MUT-GOD was found to be in the ratio 72:48:2.4, with a recovery activity INV-MUT of more than 42.9%. pH 5.8-6.5 was the most suitable range of acidity for the sensor activity. The optimum temperature was 35-45°C. The system exhibited good linearity in the range of 5 x 10^-4 approximately 10^-1 M sucrose (kinetic method) and 10^-5 approximately 2 x 10^-3 M sucrose (steady state method), in short response time (20 seconds for kinetic method, 2 minutes for steady state method), CV = 1.7% (kinetic method). The sensor had been used for determining sucrose concentration in fermentation broth, with an average recovery rate of 98%. The interference caused by the presence of glucose derived from decomposition of sucrose was eliminated by calibration with a GOD sensor. No significant loss of the enzyme electrode activity was observed after 120 hours of the continuous-flow of fresh 1 mM sucrose. The multiple-enzyme membrane showed a relatively long lifetime (compared with 14 hours as reported previously) and good storage stability (30 days, stored in distilled water at 4°C).
Fermentation broth Electrode Heated reaction Immobilized enzyme Interferences Kinetic pH Teflon membrane Steady state

"Trace Analysis Of Sugars By HPLC And Post-column Derivatization"
Chromatographia 1987 Volume 23, Issue 9 Pages 657-662
H. Engelhardt and P. Ohs

Abstract: Sugars, e.g., arabinose, glucose, fructose (each 80 ng), raffinose and sucrose (each 160 ng), were separated on a column at 36°C containing the strongly basic anion-exchange resin HPIC-AS6 (Dionex) with 0.15 M NaOH as mobile phase (0.5 mL min-1). The eluate was treated with 0.2% thymol in concentrated H2SO4 in a specially designed reaction coil operated at >90°C. Detection was at 500 nm. Results are presented for 17 sugars. The method has been applied to the determination of fructose and glucose in wine with a high residual sugar content (after a 100-fold dilution) and to dry wine, for which a 'fingerprint' analysis illustrates a separation of several sugars. Sugar alcohols, acids and other wine constituents did not interfere.
Wine HPIC Spectrophotometry Heated reaction Interferences Post-column derivatization

"Flow Stream Detectors Based On Electrocatalytic Oxidation Of Polyhydroxy Compounds At Silver Oxide Electrodes"
Contemp. Electroanal. Chem. 1990 Volume 1, Issue 1 Pages 275-296
Terrence P. Tougas, Edwin G. E. Jahngen, Michael Swartz

Abstract: Many simple carbohydrates and other polyhydroxy compounds can be oxidized at a silver oxide surface. The oxidation is via an electrocatalytic mechanism involving a Ag(I) oxide. This forms the basis of a flow stream detector operated in an amperometric mode which may be used for either flow injection or high performance liquid chromatography (HPLC) applications. The title electrode has been applied to the detection of simple carbohydrates, triglycerides and nucleic acid components.
Electrode Amperometry Detector

"Online Monitoring Of Enzyme-catalyzed Biotransformations With Biosensors"
Enzyme Microb. Technol. 1997 Volume 20, Issue 6 Pages 432-436
Frank Lammers and Thomas Scheper

Abstract: A computer-controlled flow injection analysis system with an enzyme thermistor as detector is presented for online monitoring of enzyme-catalyzed syntheses performed in a simple batch reactor with immobilized biocatalyst. Reactions studied were (1) L-arginine to L-ornithine + urea, by arginase, (2) N-acetyl-L-methionine to L-methionine, by acylase [aminoacylase, EC 3.5.1.14], and (3) fructose from sucrose, by invertase + glucose isomerase [xylose isomerase from Streptomyces albus, EC 5.3.1.5]. All the enzymes were separately immobilized from phosphate-buffered solution at 4°C, pH 7, using VA-Epoxy-Biosynth from Riedel de Haen of Seelze in Germany. The method allowed continuous monitoring and optimization of enzymatic processes.
Sensor Thermistor Immobilized enzyme Process monitoring Optimization

"Enzymic Determination Of Glucose, Sucrose And Maltose In Food Samples By Flow Injection Analysis"
Food Chem. 1990 Volume 35, Issue 2 Pages 109-116
S. M. Tzouwara-Karayanni* and S. R. Crouch

Abstract: Wheat flour (1 g) was mixed with ethanol (1 ml) and the mixture was diluted to 10 mL with water. After centrifugation (20 min at 1000 rpm), the supernatant solution was analyzed. Soft drinks were degassed, and honey was diluted (1:10) before analysis. Wine needed no pre-treatment. For determination of glucose (I), sample solution (0.1 to 1 ml) were diluted to 10 mL with 0.05 M phosphate buffer and then passed through a single bead string reactor containing glucose oxidase. The eluate from this reactor was mixed with a reagent stream containing peroxidase (0.8 mg mL-1), 1 mM 4-aminoantipyrine and 1 mM 3,5-dichloro-2-hydroxyphenylsulfonic acid in 0.05 M phosphate buffer and passed through a plain single bead string reactor. The absorbance was measured at 510 nm. For determination of sucrose (II), the samples were treated with invertase; for the determination of maltose (III) samples were treated with maltase. Calibration graphs were rectilinear from 0.01 to 0.08% of I and III and from 0.01 to 0.12% of II. Results compared well with those by the AOAC method. Recoveries were quantitative.
Wine Soft drink Spectrophotometry Immobilized enzyme Dilution Buffer Enzyme Calibration Single bead string reactor Method comparison

"Removal Of Ascorbic Acid Interference In The Determination Of Glucose And Sucrose In Non-alcoholic Beverages"
Food Chem. 1993 Volume 48, Issue 1 Pages 95-98
S. M. Tzouwara-Karayanni*, M. I. Karayannis* and S. R. Crouch*

Abstract: With use of a computer-controlled flow injection system, one portion of the degassed or (for natural orange juice) centrifuged sample was diluted with 0.05 M phosphate buffer of pH 6.86 and injected into the same buffer as carrier. The stream was passed through a single-bead string reactor containing glucose oxidase immobilized on glass beads and then mixed with Trinder reagent (containing 0.8 mg/ml of peroxidase, 1 mM 4-aminoantipyrine and 1 mM 3,5-dichloro-2-hydroxybenzenesulfonic acid). The absorbance of the product was recorded at 510 nm. A second portion of sample was treated with invertase (β-fructofuranosidase) and analyzed similarly to determine sucrose. A third portion was treated with invertase and made 40 µm in ascorbic acid (I) before analysis to determine the contribution of I. A fourth portion was treated with invertase, I and ascorbase (L-ascorbate oxidase) to remove the ascorbic acid and also to invert sucrose. Application of a standard addition method for glucose indicated recoveries in the range 99.1-102.7%, and the RSD (n = 6) for 1.1 mM glucose was 3%.
Beverage Spectrophotometry Interferences Glass beads

"Automatic Enzymatic Determination Of True Sucrose In Beet And Molasses"
Int. Sugar J. 1991 Volume 93, Issue 1110 Pages 121-125
Bengtsson, M.;Tjebbes, J.

Abstract: An outline is presented of a flow injection method in which the sample is injected into a stream of citrate buffer solution and passes through a 'glucose killer' reactor, containing immobilized aldose 1-epimerase, glucose oxidase and catalase, and then through a 'sucrose reactor' containing immobilized β-fructofuranosidase, aldose 1-epimerase and glucose oxidase. A color reagent is injected into the resulting stream to react with the H2O2 formed, and the product is detected colorimetrically. The method has been applied to digests of beet and to molasses and juices. Results were lower for cossettes than those obtained by the conventional pol analysis; the results for molasses differed only slightly from those of pol analysis and appeared to demonstrate that the correction usually applied for raffinose is too high.
Vegetable Molasses Automation Buffer Immobilized enzyme Reactor

"Flow Injection Amperometric Determination For Sucrose And Glucose In Sugarcane Juice And Molasses"
Int. Sugar J. 1998 Volume 100, Issue 1195 Pages 320-324
Luiz De Mattos, I.;Zagatto, E.A.G.;De Oliveira Neto, G.

Abstract: A flow injection system for amperometric determination of sucrose (I) and glucose (II) employing mutarotase and glucose oxidase immobilized on controlled-pore glass was developed. A Pt microelectrode polarized at 650 mV vs Ag/AgCl was used as the working electrode. The proposed system allows sequential determination of I (0-8.0 mM, r = 0.99973, RSD = 1.5%) and II (0 - 0.60 mM, R = 0.99852, RSD = 1.2%) and was applied to the anal. of sugarcane juice and molasses. Results were compared with those provided by HPLC, and the agreement between them corroborated the applicability of the proposed procedure for large scale anal. and(or) for quality control of ethanolic fermentation Sampling frequency was 50/h.
Juice Food Fermentation broth Amperometry Immobilized enzyme Controlled pore glass Method comparison

"Analysis Of Various Sugars By Means Of Immobilized Enzyme Coupled Flow Injection Analysis"
J. Biotechnol. 1996 Volume 50, Issue 2-3 Pages 93-106
B. Weigela, B Hitzmanna, G. Kretzmera, K. Sch&uuml;gerla,*, A. Huwigb and F. Giffhornb

Abstract: Glucose, maltose, sucrose, lactose, xylose, sorbose, galactose, fructose and gluconolactone were analyzed by means of immobilized pyranose oxidase as well as by the combination of immobilized glucose oxidase with immobilized glycoamylase, invertase, mutarotase, maltase (α-glucosidase) and glucose isomerase by flow injection analysis (FIA). For the simultaneous analysis of glucose and other sugars three different flow injection configurations were applied and compared. The average error of prediction of the analyzes were better than 3% in model media and better than 6% in yeast extract containing media.
Chemiluminescence Immobilized enzyme Simultaneous analysis Manifold comparison

"Simple Flow Injection Analysis System For Determination Of Added Sugars In Dairy Products"
J. Dairy Res. 1998 Volume 65, Issue 4 Pages 675-680
EDUARDO CORT&Oacute;N a and GUILLERMO LOCASCIO

Abstract: A microbial sensor based on a carbon dioxide electrode coupled with immobilized Saccharomyces cerevisiae (baker's yeast) was used for the determination of sucrose in dairy products. The sensor was used as the detector in a flow injection analysis system. Calibration curves for sucrose were established from 1 to 100 g/L. Determinations for several dairy products containing added sucrose gave good agreement with the concentrations given by manufacturers. Typically, the standard error of the method was shown to be <5% of the calculation mean.
Food Electrode Apparatus Detector

"Flow Injection Analysis System For Conductometric Measurement Of Sugar Content. Study On Electrochemical Measurement Of Sugar Content Of Food, Part VII "
Nippon Shokuhin Kagaku Kogaku Kaishi 1985 Volume 32, Issue 8 Pages 576-581
Takakazu NOMURA, Hiroyuki UKEDA, Kiyoshi MATSUMOTO, Yutaka OSAJIMA

Abstract: A conductometric flow injection analysis system for measuring sugar content in food was constructed and several component of this system were investigated. The controlled-current four-electrode method was used for conductance measurements, and then the cell constant of this system was not affected by flow rate (0-5 mL min-1). The most preferrable conditions for optimum operation of the system were as follows: mixing coil length 80 cm (i.d.=0.8 mm), injection volume 0.68 ml, flow rate 2.3 mL min-1. Over the range of 5 to 30% (w/w), the linear response was obtained and the coefficient of variation was 0.20% at 20% (w/w) sucrose for 10 successive assays. The measuring time was about 3 min for each of these assays. The sugar content in citrus juice estimated with this system agreed with that obtained from phenol-sulfuric acid method within an error of 0.20% (w/w).
Juice Food Conductometry Method comparison

"Conductometric Measurement Of Sugar Content Of Food With Flow Injection Analysis System. Study On Electrochemical Measurement Of Sugar Content Of Food, Part VIII"
Nippon Shokuhin Kagaku Kogaku Kaishi 1985 Volume 32, Issue 12 Pages 916-919
Takakazu NOMURA, Hiroyuki UKEDA, Kiyoshi MATSUMOTO, Yutaka OSAJIMA

Abstract: The rapid and convenient methods based on conductometric flow injection analysis (FIA) were proposed for measuring sugar content of apple juice and juice of sugar beet root and for measuring total solids content of cow's milk. With the FIA system, all measurements were able to carry out under the same operational conditions of the system, by use of each calibration curve for each sample -apple juice, juice of sugar beet root and cow's milk, respectively. For apple juice and clarified juice of sugar beet root with tannic acid, sugar content estimated from the proposed mehtods agreed with that obtained from phenol-sulfuric acid method with an error of 0.24%. For cow's milk homogenate, the total solids content estimated from the proposed method agreed with that obtained from gravimetric method (method of A.O.A.C.) with an error of 0.19%.
Root Juice Cows Milk Conductometry

"Approach For Conductometric Flow-injection Analysis Of Salt Content In Food"
Nippon Shokuhin Kagaku Kogaku Kaishi 1985 Volume 33, Issue 1 Pages 61-66
Kiyoshi MATSUMOTO, Koh-ichi ISHIDA, Yutaka OSAJIMA

Abstract: The four-electrode cell system described earlier (Agric. Biol. Chem., 1984, 48, 2211) was applied to enable a.c. conductometric measurement of NaCl in foods (cf. Okayama et al., Anal. Abstr., 1981, 40, 6F40) in a flow-injection system. Because the samples of soy and Worcestershire sauce had high (12 to 22%) salt content, further sample dilution in a preliminary mixing coil was required. For injections of aqueous 15% NaCl, the coefficient of variation (n = 10) was 0.22%. Up to 70 samples h-1 could be analyzed and simple detergent cleaning of the system was effective.
Food Food Conductometry Surfactant

"Simultaneous Determination Of Fructose And Glucose In Syrups By Flow Injection Spectrophotometry"
Quim. Anal. 1992 Volume 11, Issue 2 Pages 221-228
De Mattos, I.L.;Zagatto, E.A.G.

Abstract: NA
Food Spectrophotometry Redox

"Determination Of Glucose In Soft Drink And Sugar-cane Juice Employing A Multicommutation Approach In Flow System And Enzymatic Reaction"
Fresenius J. Anal. Chem. 1999 Volume 364, Issue 4 Pages 358-361
Elo&iacute;sa A. M. Kronka, Ana Paula S. Paim, B. F. Reis, Jos&eacute; L. F. Costa Lima, Rui A. Lapa

Abstract: A flow system based on a multicommutation approach was developed for the determination of glucose and sucrose employing enzymatic reactions. The determination was based on the reaction with D-glucose generating hydrogen peroxide catalyzed by glucose-oxidase (GOD). Subsequently, the H2O2 generated reacts with 4-aminefenazone plus phenol to form 4-(p-benzoquinone-mono-imine) fenazone detected at 510 nm. This reaction is catalyzed by the peroxidase enzyme (POD). The flow network comprised a set of three-way solenoid valves and was controlled by means of a microcomputer furnished with an electronic interface and running a software written in Quick BASIC 4.5. The flow network and control software were designed to implement the multicommutation approach providing facilities to handle sample and reagent solutions, so that, sample dilution could be easily performed on line. Accuracy was assessed by comparison with results obtained by known procedures and no significant difference at the 95% confidence level was observed. Other advantageous features were a linear response ranging from 0.05 to 0.20% (w/v) glucose without prior dilution, a reagent consumption of 336 µL per determination, an analytical throughput of 30 samples per hour.
Soft drink Juice Spectrophotometry Multicommutation Immobilized enzyme Computer Method comparison