Rate Of Change In Absorbance Research Articles

Application of the fuzzy proportional integral differential (PID) temperature control algorithm in a liver function test system based on a centrifugal microfluidic device

Clinical laboratory examinations frequently include biochemical analysis of the liver. The presence of alanine aminotransferase (ALT) and aspartate transaminase (AST) in serum can be used to identify liver damage. In this study, a centrifugal microfluidic-based clinical biochemical detection system was developed for the detection of liver function markers. Using the centrifugal microfluidic chip and centrifugal force on the chip, separation of blood cells and serum was performed. The extraction and mixing of quantitative serum and diluent were completed under the chip design of microchannels and microchambers. The lyophilized reagent beads in the chip interacted with the combined solution. The Fuzzy PID algorithm regulates the power of the heating film to deliver the ideal reaction temperature. In accordance with Beer–Lambert, the rate of change in the absorbance of the reaction solution at 340 nm of the light source was measured and a standard curve for the relationship between concentration and rate of change in absorbance was constructed. The system is portable, quick, and simple to use because it uses a centrifugal microfluidic chip instead of the conventional detection and analysis approach. In the future, it is anticipated that the system will have several applications in the detection of highly integrated on-chip point-of-care devices.

Effect of Protein Hydrolysates from Limonia (L.) acidissima and Citrullus (C.) lanatus on Anthocyanin Degradation

ABSTRACT Anthocyanins are natural plant pigments and used extensively in many food and beverage formulations due to their atrractive red, blue, or purple color. However, this color is highly sensitive to light, and therefore unstable and degraded even in the presence of ascorbic acid. This leads to the loss in the bioactivity of anthocyanin and also loss in the color of food or beverages. Hence, the present study focuses on the preservation of anthocyanin from Syzygium cumini (L.) using natural preservatives such as Limonia acidissima (L. acidissima) and Citrullus lanatus (C. lanatus) seed protein hydrolyzates at elevated temperature (35°C/5 days). Different concentrations (0.04–2%) of L. acidissima and C. lanatus protein hydrolyzates were examined for its preservative action against 5% crude extract of anthocyanin from Syzygium cumini (L.) preservation. This concentration of protein hydrolyzates followed the first order of reaction. Both concentrations of protein hydrolyzates reduced the rate of change in absorbance. However, 2% concentration of C. lanatus protein hydrolyzate was found to be very effective in providing highestthe preservation of color. Therefore, the overall study highlights the preservation effect of L. acidissima and C. lanatus protein hydrolyzate.

Absorbance characteristics of a liquid-phase gas sensor based on gas-permeable liquid core waveguides

The absorbance characteristics and influential factors on these characteristics for a liquid-phase gas sensor, which is based on gas–permeable liquid core waveguides (LCWs), are studied from theoretical and experimental viewpoints in this paper. According to theory, it is predicted that absorbance is proportional to the analyte concentration, sampling time, analyte diffusion coefficient, and geometric factor of this device when the depletion layer of the analyte is ignored. The experimental results are in agreement with the theoretical hypothesis. According to the experimental results, absorbance is time-dependent and increasing linearly over time after the requisite response time with a linear correlation coefficient r2>0.999. In the linear region, the rate of absorbance change (RAC) indicates improved linearity with sample concentration and a relative higher sensitivity than instantaneous absorbance does. By using a core liquid that is more affinitive to the analyte, reducing wall thickness and the inner diameter of the tubing, or increasing sample flow rate limitedly, the response time can be decreased and the sensitivity can be increased. However, increasing the LCW length can only enhance sensitivity and has no effect on response time. For liquid phase detection, there is a maximum flow rate, and the absorbance will decrease beyond the stated limit. Under experimental conditions, hexane as the LCW core solvent, a tubing wall thickness of 0.1mm, a length of 10cm, and a flow rate of 12mLmin−1, the detection results for the aqueous benzene sample demonstrate a response time of 4min. Additionally, the standard curve for the RAC versus concentration is RAC=0.0267c+0.0351 (AUmin−1), with r2=0.9922 within concentrations of 0.5–3.0mgL−1. The relative error for 0.5mgL−1 benzene (n=6) is 7.4±3.7%, and the LOD is 0.04mgL−1. This research can provide theoretical and practical guides for liquid–phase gas sensor design and development based on a gas-permeable Teflon AF 2400 LCW.

Apposite Insulin-like Growth Factor (IGF) Receptor Glycosylation Is Critical to the Maintenance of Vascular Smooth Muscle Phenotype in the Presence of Factors Promoting Osteogenic Differentiation and Mineralization

Vascular calcification is strongly linked with increased morbidity and mortality from cardiovascular disease. Vascular calcification is an active cell-mediated process that involves the differentiation of vascular smooth muscle cells (VSMCs) to an osteoblast-like phenotype. Several inhibitors of this process have been identified, including insulin-like growth factor-I (IGF-I). In this study, we examined the role of the IGF receptor (IGFR) and the importance of IGFR glycosylation in the maintenance of the VSMC phenotype in the face of factors known to promote osteogenic conversion. IGF-I (25 ng/ml) significantly protected VSMCs from β-glycerophosphate-induced osteogenic differentiation (p < 0.005) and mineral deposition (p < 0.01). Mevalonic acid depletion (induced by 100 nm cerivastatin) significantly inhibited these IGF protective effects (p < 0.01). Mevalonic acid depletion impaired IGFR processing, decreased the expression of mature IGFRs at the cell surface, and inhibited the downstream activation of Akt and MAPK. Inhibitors of N-linked glycosylation (tunicamycin, deoxymannojirimycin, and deoxynojirimycin) also markedly attenuated the inhibitory effect of IGF-I on β-glycerophosphate-induced mineralization (p < 0.05) and activation of Akt and MAPK. These results demonstrate that alterations in the glycosylation of the IGFR disrupt the ability of IGF-I to protect against the osteogenic differentiation and mineralization of VSMCs by several interrelated mechanisms: decreased IGFR processing, reduced IGFR cell-surface expression, and reduced downstream signaling via the Akt and MAPK pathways. IGF-I thus occupies a critical position in the maintenance of normal VSMC phenotype and protection from factors known to stimulate vascular calcification.

Major Urinary Protein-1 Increases Energy Expenditure and Improves Glucose Intolerance through Enhancing Mitochondrial Function in Skeletal Muscle of Diabetic Mice

Major urinary protein-1 (MUP-1) is a low molecular weight secreted protein produced predominantly from the liver. Structurally it belongs to the lipocalin family, which carries small hydrophobic ligands such as pheromones. However, the physiological functions of MUP-1 remain poorly understood. Here we provide evidence demonstrating that MUP-1 is an important player in regulating energy expenditure and metabolism in mice. Both microarray and real-time PCR analysis demonstrated that the MUP-1 mRNA abundance in the liver of db/db obese mice was reduced by approximately 30-fold compared with their lean littermates, whereas this change was partially reversed by treatment with the insulin-sensitizing drug rosiglitazone. In both dietary and genetic obese mice, the circulating concentrations of MUP-1 were markedly decreased compared with the lean controls. Chronic elevation of circulating MUP-1 in db/db mice, using an osmotic pump-based protein delivery system, increased energy expenditure and locomotor activity, raised core body temperature, and decreased glucose intolerance as well as insulin resistance. At the molecular level, MUP-1-mediated improvement in metabolic profiles was accompanied by increased expression of genes involved in mitochondrial biogenesis, elevated mitochondrial oxidative capacity, decreased triglyceride accumulation, and enhanced insulin-evoked Akt signaling in skeletal muscle but not in liver. Altogether, these findings raise the possibility that MUP-1 deficiency might contribute to the metabolic dysregulation in obese/diabetic mice, and suggest that the beneficial metabolic effects of MUP-1 are attributed in part to its ability in increasing mitochondrial function in skeletal muscle.

Oxygen Evolution Catalysis by a Dimanganese Complex and Its Relation to Photosynthetic Water Oxidation

[Mn2(III/IV)(mu-O) 2(terpy)2(OH 2)2](NO3)3 (1, where terpy = 2,2':6'2''-terpyridine) acts as a water-oxidation catalyst with HSO5(-) as the primary oxidant in aqueous solution and, thus, provides a model system for the oxygen-evolving complex of photosystem II (Limburg, J.; et al. J. Am. Chem. Soc. 2001, 123, 423-430). The majority of the starting [Mn2(III/IV)(mu-O)2](3+) complex is converted to the[Mn2(IV/IV)(mu-O)2](4+) form (2) during this reaction (Chen, H.; et al. Inorg. Chem. 2007, 46, 34-43). Here, we have used stopped-flow UV-visible spectroscopy to monitor UV-visible absorbance changes accompanying the conversion of 1 to 2 by HSO5(-). With excess HSO5(-), the rate of absorbance change was found to be first-order in [1] and nearly zero-order in [HSO5(-)]. At relatively low [HSO5(-)], the change of absorbance with time is distinctly biphasic. The observed concentration dependences are interpreted in terms of a model involving the two-electron oxidation of 1 by HSO5(-), followed by the rapid reaction of the two-electron-oxidized intermediate with another molecule of 1 to give two molecules of 2. In order to rationalize biphasic behavior at low [HSO5(-)], we propose a difference in reactivity of the [Mn2(III/)(IV)(mu-O)2](3+) complex upon binding of HSO5(-) to the Mn(III) site as compared to the reactivity upon binding HSO5(-) to the Mn(IV) site. The kinetic distinctness of the Mn(III) and Mn(IV) sites allows us to estimate upper limits for the rates of intramolecular electron transfer and terminal ligand exchange between these sites. The proposed mechanism leads to insights on the optimization of 1 as a water-oxidation catalyst. The rates of terminal ligand exchange and electron transfer between oxo-bridged Mn atoms in the oxygen-evolving complex of photosystem II are discussed in light of these results.

Interactions of Human Rad54 Protein with Branched DNA Molecules

The Rad54 protein plays an important role during homologous recombination in eukaryotes. The protein belongs to the Swi2/Snf2 family of ATP-dependent DNA translocases. We previously showed that yeast and human Rad54 (hRad54) specifically bind to Holliday junctions and promote branch migration. Here we examined the minimal DNA structural requirements for optimal hRad54 ATPase and branch migration activity. Although a 12-bp double-stranded DNA region of branched DNA is sufficient to induce ATPase activity, the minimal substrate that gave rise to optimal stimulation of the ATP hydrolysis rate consisted of two short double-stranded DNA arms, 15 bp each, combined with a 45-nucleotide single-stranded DNA branch. We showed that hRad54 binds preferentially to the open and not to the stacked conformation of branched DNA. Stoichiometric titration of hRad54 revealed formation of two types of hRad54 complexes with branched DNA substrates. The first of them, a dimer, is responsible for the ATPase activity of the protein. However, branch migration activity requires a significantly higher stoichiometry of hRad54, approximately 10 +/- 2 protein monomers/DNA molecule. This pleomorphism of hRad54 in formation of oligomeric complexes with DNA may correspond to multiple functions of the protein in homologous recombination.

Reduction of Carboxylic Acids by Nocardia Aldehyde Oxidoreductase Requires a Phosphopantetheinylated Enzyme

Aldehyde oxidoreductase (carboxylic acid reductase (Car)) catalyzes the magnesium-, ATP-, and NADPH-dependent reduction of carboxylic acids to their corresponding aldehydes. Heterologous expression of the car gene in Escherichia coli afforded purified recombinant enzyme with a specific activity nearly 50-fold lower than that of purified native Nocardia sp. enzyme. The 5-fold increase in specific activity obtained by incubating purified recombinant Car with CoA and Nocardia cell-free extracts indicated that post-translational phosphopantetheinylation of Car is required for maximum enzyme activity. Nocardia phosphopantetheine transferase (PPTase) expressed in E. coli was isolated and characterized. When incubated with [(3)H]acetyl-CoA and Nocardia PPTase, the labeled acetylphosphopantetheine moiety was incorporated into recombinant Car. Coexpression of Nocardia Car and PPTase in E. coli gave a reductase with nearly 20-fold higher specific activity. Site-directed mutagenesis in which Ser(689) was replaced with Ala resulted in an inactive Car mutant. The results show that Car expressed in Escherichia coli is an apoenzyme that is converted to a holoenzyme by post-translational modification via phosphopantetheinylation. Doubly recombinant resting E. coli cells efficiently reduce vanillic acid to vanillin.

Insulin-like Growth Factor-binding Protein-5 Activates Plasminogen by Interaction with Tissue Plasminogen Activator, Independently of Its Ability to Bind to Plasminogen Activator Inhibitor-1, Insulin-like Growth Factor-I, or Heparin

Transgenic mice expressing IGFBP-5 in the mammary gland exhibit increased cell death and plasmin generation. Because IGFBP-5 has been reported to bind to plasminogen activator inhibitor-1 (PAI-1), we determined the effects of this interaction in HC11 cells. PAI-1 prevented plasmin generation from plasminogen and inhibited cleavage of focal adhesions, expression of caspase 3, and cell death. IGFBP-5 could in turn prevent the effects of PAI-1. IGFBP-5 mutants with reduced affinity for IGF-I (N-term) or deficient in heparin binding (HEP- and C-term E and F) were also effective. This was surprising because IGFBP-5 reportedly interacts with PAI-1 via its heparin-binding domain. Biosensor analysis confirmed that, although wild-type IGFBP-5 and N-term both bound to PAI-1, the C-term E had greatly decreased interaction with PAI-1. This suggests that IGFBP-5 does not antagonize the actions of PAI-1 by a direct molecular interaction. In a cell-free system, using tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA) to activate plasminogen, PAI-1 inhibited plasmin generation induced by both activators, whereas IGFBP-5 prevented the effects of PAI-1 on tPA but not uPA. Furthermore, we noted that IGFBP-5 activated plasminogen to a greater extent than could be explained solely by inhibition of PAI-1, suggesting that IGFBP-5 could directly activate tPA. Indeed, IGFBP-5 and the C-term E and F were all able to enhance the activity of tPA but not uPA. These data demonstrate that IGFBP-5 can enhance the activity of tPA and that this can result in cell death induced by cleavage of focal adhesions. Thus IGFBP-5 can induce cell death by both sequestering IGF-I and enhancing plasmin generation.

Quantitative determination of heparin levels in serum with microtiter plate-format optode

A new assay method has been developed for the quantitative determination of heparin in serum using a microtiter plate-format optode (MPO). Heparin and proton in physiological sample are favorably co-extracted into the solvent polymeric optode membrane containing both cationic lipophilic additive, tridodecylmethyl ammonium chloride (TDMAC), and proton-selective ionophore, 3-hydroxy-4-(4-nitrophenylazo)-phenyloctadecanoate (ETH 2412), resulting in the absorbance change of the membrane to varying heparin levels. The optimized MPO composition contains low polymer-to-plasticizer ratio compared to those of conventional ion-selective optodes or electrodes, i.e., poly(vinyl chloride) (20.0)/dioctylsebacate (76.3)/ETH 2412 (1.7)/TDMAC (1.0) (wt.%): it resulted in a quantitative response to heparin from 0 to 15 unit/mL in serum with high sensitivity. The heparin–protamine titration on the MPO could provide rapid and precise determination of heparin. It was shown that the heparin levels in serum sample could be determined from the rate of absorbance change over time (Δ A/Δ t); this method was more effective than the direct absorbance measurement in minimizing the interferences from color and turbidity of serum samples. MPO has been developed as a high throughput and convenient disposable sensing device, and may find a wide application in the determination of polyions and charged macromolecules.

PAR1 Cleavage and Signaling in Response to Activated Protein C and Thrombin

Activated protein C (APC), a natural anticoagulant protease, can trigger cellular responses via protease-activated receptor-1 (PAR1), a G protein-coupled receptor for thrombin. Whether this phenomenon contributes to the physiological effects of APC is unknown. Toward answering this question, we compared the kinetics of PAR1 cleavage on endothelial cells by APC versus thrombin. APC did cleave PAR1 on the endothelial surface, and antibodies to the endothelial protein C receptor inhibited such cleavage. Importantly, however, APC was approximately 10(4)-fold less potent than thrombin in this setting. APC and thrombin both triggered PAR1-mediated responses in endothelial cells including expression of antiapoptotic (tumor necrosis factor-alpha-induced a20 and iap-1) and chemokine (interleukin-8 (il-8) and cxcl3) genes, but again, APC was approximately 10(4)-fold less potent than thrombin. The addition of zymogen protein C to endothelial cultures did not alter the rate of PAR1 cleavage at low or high concentrations of thrombin, and PAR1 cleavage was substantial at thrombin concentrations too low to trigger detectable conversion of protein C to APC. Thus, locally generated APC did not contribute to PAR1 cleavage beyond that effected by thrombin in this system. Although consistent with reports that sufficiently high concentrations of APC can cleave and activate PAR1 in culture, our data suggest that a significant physiological role for PAR1 activation by APC is unlikely.

Catalytic Kinetic Determination of Silver through its Catalytic Effect on Congo Red-Peroxodisulphate Reaction

A kinetic-spectrophotometric procedure is reported for determination of trace amounts of Ag(I). This method is based on catalytic effect of silver ion on the oxidation of congo red by potassium peroxodisulphate at pH=3.4 and 30 oC in the presence of 1,10-phenanthroline as an activator. The reaction was followed spectrophotometrically by measuring the rate of change in absorbance of congo red at 510 nm in the range of 30-330 seconds after initiation of the reaction. Silver can be determined in the range 2-100 and 100-1100 ng/ml with detection limit 1 ng/ml. Ten replicate analysis of a sample solution containing 8 ng/ml silver gave a relative standard deviation of 1.0%. This method was used for determination of silver in water and photographic plate samples.

Micro Determination of Ammonia by a Kinetic Spectrophotometric Method

A simple and sensitive kinetic spectrophotometric method is described for the determination of micro amounts of ammonia. The method is based on the inhibiting effect of ammonia on the oxidation rate of aqua ethylenediamine-tetraacetate chromium(III) [Cr(HEDTA)(H2O)] with N-bromo succinimide (NBS) to yield chromium(VI). The reaction was followed spectrophotometrically by measuring the rate of change in absorbance with time at 365 nm, which is the maximum absorption wavelength of chromium(VI). Using the recommended procedure, ammonia can be determined up to 153 µg ml−1 with a linear calibration graph and a detection limit of 0.09 µg ml−1. The method was successfully applied to the determination of ammonia in waste water samples. Recovery experiments for the analyzed samples were 97–100% with relative standard deviation, Sr (%) of ≤ 2.9% indicating high accuracy and precision of the proposed method. The interference of various cations and anions in the determination of ammonia was investigated.

Catalytic Kinetic Determination of Ultratrace Amounts of Silver With Spectrophotometric Detection

ABSTRACT Silver has a very strong catalytic effect on the peroxodisulphate oxidation of Brilliant Cresyl Blue in the presence of 1.10-Phenanthroline as an activator. The reaction was followed spectrophotometrically by measuring the rate of change in absorbance of brilliant cresyl blue at 635 nm and 30°C. Silver can be determined in the range of 0.3 – 1500.0 ng/ml by a fixed time method. The limit of detection is about 0.1 ng/ml of silver. Silver in real samples was determined with satisfactory results.

Mechanism and elimination of aspirin-induced interference in Emit II d.a.u. assays

The presence of salicylates in urine reduces the signal in Emit assays (Syva), potentially yielding false-negative drugs-of-abuse screening results. We demonstrate that the principal urinary metabolite of salicylate, salicyluric acid (SUA; 2-hydroxybenzoylaminoacetic acid), interferes with the measurement of NADH formed in the assay by reducing the molar absorptivity of NADH at 340 nm. Thus, for a given concentration of d.a.u. analyte the change in absorbance over the assay time interval is less in the presence of SUA. With the Emit cocaine assay on the Hitachi 704 analyzer, the rate of absorbance change (delta AR) monitored at 340 nm for a specimen containing approximately 270 micrograms/L benzoylecgonine (BE) was 57 +/- 1.9 mA/min without SUA and 29 +/- 2.7 mA/min with 5 g/L SUA (n = 20). In contrast, delta AR determined at 376 nm was 18.6 +/- 0.5 mA/min with and 17.9 +/- 0.8 mA/min without 5 g/L SUA (n = 20). Measuring the Emit assay signal at wavelengths where SUA has no absorbance (376 nm) eliminates the interference due to SUA while maintaining the precision of the assay near the cutoff concentration for BE (300 micrograms/L).

Kinetic-Spectrophotometric Determination of Traces of Osmium by Its Catalytic Effect on the Oxidation of Pyrogallol Red by Hydrogen Peroxide

Abstract A Kinetic-spectrophotometric method for the determination of ultra-trace amounts of osmium(VIII) is described. It is based on the catalytic action of osmium(VIII) on the oxidation of pyrogallol red with hydrogen peroxide, yielding a colorless product in neutral medium. The reaction is followed spectrophotometrically by measuring the rate of change in absorbance at 540 nm and 30°C. Os(VIII) in the range 0.005 -100 ng.ml−1 can be determined. The proposed method is hardly subject to interferences. The relative standard deviation is 1.5% for 1 ng.ml−1 of Os(VIII). The kinetic parameters of the catalyzed and uncatalyzed reactions are reported. The detection limit is 1 pg.ml−1.

Simplified enzymatic assay of angiotensin-converting enzyme in serum

We report a simple, enzymatic method for determining angiotensin-converting enzyme (ACE; EC 3.4.15.1) in serum. The proposed method features coupling an established reaction catalyzed by gamma-glutamyltransferase (GGT; EC 2.3.2.2) to the ACE reaction, which releases glycylglycine from the artificial substrate hippuryl-glycyl-glycine. The glycyl-glycine released by the ACE reaction becomes rate-limiting in the GGT reaction, in which it participates as a receptor substrate for a gamma-glutamyl group transferred from a donor substrate, L-gamma-glutamyl-3-carboxy-4-nitroanilide. The reaction releases 3-carboxy-4-nitroaniline, which is monitored spectrophotometrically at 410 nm. The resulting rate of change in absorbance is linearly related to the glycyl-glycine concentration and therefore to the activity of ACE. The linear range of the method extends from ACE values < 50 to at least 1300 U/L of serum. Good precision is indicated by a low CV for replicate analyses (3.6% and 4.6% for within-run and day-to-day assays, respectively, for normal ACE activity, and 3.1% within-run for high ACE activity). Results also correlate well with those of an established colorimetric method (r = 0.978). The major advantages of the method are its procedural simplicity, limited cost, use of readily available reagents, applicability to isoenzyme studies, and adaptability to automation.

Kinetic-spectrophotometric determination of molybdenum (VI) and tungsten (VI) in mixtures

A simple kinetic-spectrophotometric method is described for the determination of molybdenum(VI) and tungsten(VI) in mixtures, without prior separation. The method is based on the catalytic effect of molybdenum(VI) and tungsten(VI) on the oxidation of 2,4-diaminophenol dihydrochloride (DAP) by hydrogen peroxide in acidic medium. The reaction was followed spectrophotometrically by measuring the rate of change in absorbance with time at 500 nm. A partial inhibition in the catalytic activity of each catalyst, when the other one is present, at all ratios of Mo(VI) ∶ W(VI) mixtures studied was observed. On the other hand, the catalytic activity of tungsten(VI) dropped to zero whilst that of molybdenum(VI) decreased slightly, in the presence of citrate ions. Two sets of experiments were carried out, the first in the absence and the other in the presence of citrate, and the resolution of Mo(VI)/W(VI) mixtures was achieved by solving two simultaneous equations. Various molar ratios of Mo(VI) ∶ W(VI), at the 10−6M level, from 0.2 ∶ 1 to 5 ∶ 1 can be determined with satisfactory precision and accuracy. The selectivity of the method was investigated and the method was applied successfully to the determination of molybdenum and tungsten in each other's presence in steel.

Spectrophotometric Determination of Some Dibenzazepines

Abstract Simple and sensitive method is proposed for determination of four di-benzazepine derivatives. The method is based on the oxidation of dibenzazepines by potassium dichromate in acid medium. The reaction was followed spectrophoto-metrically by measuring the rate of change in absorbance of the coloured products at 670 nm. Linear calibration graphs from 5-50 μg/ml of the final solutions are obtained for all the studied drugs. The method has been successfully applied to the determination of dibenzazepines in some commercial and laboratory prepared tablets. The results were compared statistically with those obtained by official procedures.

Further studies on autoclaved‐induced toxicity in tissue culture media: gauging sugar breakdown by spectrophotometry

Fructose and sucrose were used to investigate cyanide‐induced absorption changes after high temperature treatment. By comparing the time‐resolved absorbance difference spectra obtained under aerobic conditions with those under aerobic conditions, absorbance changes that are associated with the process of oxygen consumption were identified. The rates of absorbance changes of autoclaved sucrose solution or of autoclaved MS medium (Murashige and Skoog 1962. Physiol. Plant. 15: 473–497) were correlated with those of oxygen consumption measured by polarography and used for determining the toxicity of culture media (Hsiao and Bornman 1991. Physiol. Plant. 81: 55–58). When autoclaved together with sucrose, FeNa‐EDTA promotes its degradation. Absorbance change, therefore, is a convenient parameter for measuring not only the extent of carbohydrate breakdown at high temperature but also the relative toxicity of culture media autoclaved under different conditions.