Concentrations Of Triton X-100 Research Articles

Reverse flotation of non-coking coal fines using non-ionic surfactant triton X-100 as depressant

A novel approach to apply Triton X-100 (TX-100) surfactant as depressant in reverse flotation of non-coking coal fines, which is different from the conventional use of dextrin as an depressant. Fourier transformed infrared spectroscopy (FTIR), contact angle and induction time instruments were used to analyze the mechanism of high-concentration TX-100 depressing the floating of coal particle. FTIR results revealed that TX-100 could physically adsorb on the surface of coal. Contact angle results showed the higher concentration of TX-100 aqueous solution was more likely to wet the surface of TX-100-treated coal, slowing down the film thinning and rupture rate during bubble-coal adhesion. In high-concentration TX-100 aqueous solution, the bubble surface was modified by the coverage of TX-100 molecules, which makes bubble difficult to adhere to coal particle. Induction time results showed the adhesion probability of coal particle and bubble gradually decreases with the increase of TX-100 concentration.

Immobilisation of Candida rugosa lipase on a highly hydrophobic support: A stable immobilised lipase suitable for non-aqueous synthesis

Lipase from Candida rugosa (CrL) was immobilised on highly hydrophobic, octadecyl methacrylate resin (Lifetech™ ECR8806M) via interfacial adsorption. The aim was to produce a stable biocatalyst suitable for use in a range of lipid-modifying reactions. Immobilisation was carried out in 10 mM phosphate buffer (pH 6.0) over 24 h at 21 °C. High protein binding of 58.7 ± 4.9 mg/g dry support accounted for ∼53 % of the applied protein. The activity recovery against tributyrin was 74.0 ± 1.1 %. The specific activity of immobilised CrL against tributyrin was considerably higher than that of Novozym® 435, at 1.79 ± 0.05 and 1.08 ± 0.04 U/mg bound protein, respectively. Incubation with high concentrations (10 % w/v) of both Triton X-100 and SDS resulted in only a small reduction in immobilised lipase activity. Solvent-free synthesis of glycerides by the FFA-saturated immobilised CrL was successful over 6 reaction cycles, with no apparent loss of activity.

Cloud Point Extraction for Pre-Concentration and Spectrophotometric Determination of Trace Amounts of Silver Ions

During the last years, the cloud point extraction was applied for determination, separation, and enrichment of the elements. The current study used cloud point extraction (CPE) for the extraction of Ag(I) ions pre-concentration from watery solutions by a non-ionic surfactant (Triton X-114) and chelating agent a 6-(4-bromo-phenylazo)m-anisidine[6-(4-BrPAA)], then estimation by using the spectrophotometry at 514 nm. Several condition effects on the efficiency of the cloud-point extraction included Triton X-114 concentration, [6-(4-BrPAA)] concentration, pH, time, and incubation temperature. The silver reacts with [6-(4-BrPAA)] to produce complex at a ratio of one to one. 0.009 to 1.5 μg mL-1 is the range of linearity. The detection limit and quantification of Ag(I) ion were 0.0054 and 0.0182 μg mL-1, respectively. The interference of the cations was examined. The cloud-point extraction was used for the evaluation of silver concentration in the water specimen.

Triton Ziehl-Neelsen technique does not improve detection of TB bacilli in sputum

Background: Addition of Triton X 100 reagent to conventional Ziehl-Neelsen staining has been shown to improve detection of tuberculosis bacilli in CSF to diagnose TB meningitis. We evaluated its performance in sputum in the diagnosis of pulmonary tuberculosis. Methods: We compared the Triton ZN technique with the conventional ZN method on the sputum of 68 patients suspected with pulmonary tuberculosis by Bland Altman agreement analysis. Results: The Triton ZN technique detected lower number of bacilli than the conventional AFB staining method at all the tested concentrations of Triton X100 by a mean difference in the AFB score ranging from 0.2 to 0.5. Conclusions: Our study suggests that Triton X 100 added to conventional ZN staining does not improve detection of M. tuberculosis in sputum.

Mutations in the Reelin pathway are associated with abnormal expression of microglial IgG FC receptors in the cerebellar cortex.

Microglia are the immune cells of the central nervous system involved in a variety of developmental processes, such as regulation of cell death and survival, spatial patterning, and contribute to the development of Purkinje cells (PCs) during migration. Microglia express immunoglobulin G Fc receptors (FcgRs). In this report, we describe microglial FcgR expression and its relation to abnormal PC migration in the cerebellum during development. To detect microglial FcgR, the direct anti-IgG (secondary antisera) and high concentrations of Triton X-100 were applied as a method for labeling microglial cells without the use of any specific primary antiserum. By using Acp2-/- mice, which show an excessive PC migration into the molecular layer (ml), and 3 different types of mice with a null to alter the Reelin pathway (Reeler-, Dab1 (SCM)-, and Apoer mutant mice), we studied the location of PCs and the expression of FcgRs. Wild type littermates were used as controls in all studies. We show that the expression of microglial FcgRs was absent and PCs were ectopically located in the white matter in the cerebella of all mutant mice, except for the Acp2-/- mice (PCs were located in the ml). These results suggest a role for FcgRs in the Reelin signaling pathway, not in regulating PC migration, but rather in the adaptation to an environment with a relatively large number of ectopically located PCs. However, the exact correlation between the ectopic location of PCs and lack of FcgRs in Reeler, SCM, and Apoer-/- mice and the presence of FcgRs and directed PC location in the ml in Acp2-/- mice are yet to be determined.

Molecular Expression of Bioactive Recombinant Methionine Sulfoxide Reductase A (MsrA).

The increase in reactive oxygen species (ROS) production during cryopreservation of semen, leads to oxidation of biomolecules affecting the functionality of spermatozoa. Methionine residues in proteins are highly prone to oxidation and get converted into methionine sulfoxide (MetO). Methionine sulfoxide reductase A (MsrA) can improve the functionality of spermatozoa by reducing the MetO to methionine restoring the lost functionality of the affected proteins. The expression of catalytically active recombinant MsrA (rMsrA). The msrA gene was PCR amplified, cloned and sequenced. Further, the recombinant clone was used for protein expression and purification. The protein was getting precipitated during dialysis in Tris-buffer. Hence, the purified rMsrA was dialyzed at 4°C against the Tris-buffer pH 7.5 containing MgCl2, KCl, NaCl, urea and triton X-100. During dialysis, changes of buffer were done at every 12 h interval with stepwise reduction in the concentrations of NaCl, urea and triton X-100. The final dialysis was done with buffer containing 10 mM MgCl2, 30 mM KCl, and 150 mM NaCl, 25 mM Tris-HCl pH 7.5. The activity of the rMsrA was checked spectrophotometrically. The protein BLAST of buffalo MsrA with bovine sequence showed 14 amino acid mismatches. The rMsrA has been purified under denaturing conditions as it was forming inclusion bodies consistently during protein expression. After renaturation, the purified 33 kDa rMsrA was catalytically active by biochemical assay. The rMsrA expressed in prokaryotic system is catalytically active and can be used for supplementation to semen extender to repair the oxidatively damaged seminal plasma proteins that occur during cryopreservation.

Adsorption onto MWCNTs Coupled with Cloud Point Extraction for Dye Removal from Aqueous Solutions: Optimization by Experimental Design.

The main aim of the study was to examine the feasibility and benefits of adsorption onto multi-walled carbon nanotubes (MWCNTs) coupled with cloud point extraction (CPE) for the removal of Rhodamine B (RB) from aqueous solutions. MWCNTs offer the particular features of the ideal adsorbents for the organic dyes such as hollow tubular structure and specific surface area. Nevertheless, they suffer from the drawbacks of low dispersion in the aqueous solutions and separation inconvenience from the media. Cloud point extraction, combined with the adsorption onto MWCNTs can be a promising method to overcome the problems. In the study, adsorption onto MWCNTs coupled with CPE was applied for RB removal from aqueous solutions. The process was optimized by the response surface modeling method. Moreover, the applicability of the proposed method in the real sample analyses was investigated. MWCNTs were used as adsorbent and Triton X-100 (TX-100) as the nonionic surfactant for CPE process. The experiments were carried out based on a Box-Behnken design (BBD) with the input variables of MWCNTs dosage (0.6-1.2 mg), solution pH (3-9), clouding time (20-40 min) and TX-100 concentration (10-20 v/v%) using 5 mg L-1 RB solutions. Regression analyses resulted in a statistically significant quadratic model (R2=0.9718, F=24.96, p<0.0001) by which the optimum levels of the variables were predicted as: MWCNTs dosage of 0.7 mg, pH=3, clouding time of 39.9 minutes and TX-100 concentration of 19.91% (v/v). The predicted conditions were experimentally validated by achieving an RB removal of 94.24%. Based on the results, the combination of the environmentally friendly technique of CPE with adsorption onto MWCNTs allows the efficient removal of RB from water samples and the method can be effectively optimized by the response surface modeling.

МОНІТОРИНГ ВМІСТУ НАТРІЮ, КАЛІЮ, КАЛЬЦІЮ ТА МАГНІЮ В ПРОДУКТАХ ПЕРЕРОБКИ МОЛОЧНОЇ СИРОВАТКИ

The influence of Triton X-100 concentration and time of ultrasonic treatment (frequency 44 kHz, intensity 1.5 W/cm2) on the determination of sodium, potassium, calcium and magnesium in whey processed products by atomic absorption spectroscopy (AAS) was studied. It was shown that the maximum value of the analytical signal is achieved with a mass fraction of Triton X-100 of 5 % and an ultrasonic treatment time of 15 min. The sensitivity of measurements increases from one and a half to two times. Procedures have been developed for determining the content of sodium, potassium, calcium and magnesium in whey and products of its processing by AAS and atomic emission spectroscopy with inductively coupled plasma (AES-ICP). Validation of the results of determination of magnesium and calcium by AAS and AES-ICP was carried out by varying the weight of the sample and standard addition method.

Extraction induced by emulsion breaking for simultaneous determination of Co, Fe and Ni in petroleum asphalt cement by high-resolution continuum source atomic absorption spectrometry

This paper reports, for the first time, the simultaneous determination of Co, Fe and Ni in petroleum asphalt cement by high-resolution continuum source atomic absorption spectrometry after extraction induced by the emulsion breaking method. The variables that could influence the sample preparation were evaluated, like sample dilution with toluene and mineral oil, extraction time, HNO3 and Triton X-100 concentration in the aqueous extractant solution. At optimized conditions, 0.08 g of the sample was dissolved with 1.5 mL of toluene and 3.5 mL of mineral oil and vigorous mixed with the extractant solution containing 40% (v/v) of HNO3 and 40% (v/v) of Triton-X-100. The emulsions were broken by heating just after their formation and the aqueous extracts were collected for analysis. Simultaneous determination was carried out at the secondary lines of Co (352.685 nm), Fe (352.604 nm) and Ni (352.454 nm). For this purpose, the best pyrolysis and atomization temperatures were 700 and 2400 °C, respectively. Calibration was performed with standard aqueous solutions and no matrix effects were observed. The limit of detection was 0.003 mg L−1 for Co, 0.018 mg L−1 for Fe and 0.027 mg L−1 for Ni. Method accuracy was confirmed by comparing the results obtained by the proposed extraction method with those obtained after samples acid digestion. Recoveries ranged from 100 to 107%, 103 to 109% and 95.5 to 98.9% were obtained for Co, Fe and Ni, respectively.

Optimization of aqueous two-phase micellar system for partial purification of L-asparaginase from Penicillium sp. grown in wheat bran as agro-industrial residue.

L-asparaginase has been used in the remission of malignant neoplasms such as acute lymphoblastic leukemia. The search for new sources of this enzyme has become attractive for therapeutics. Traditional methods for biomolecule purification involve several steps. A two-phase system may be a good strategy to anticipate one of these stages. This study aimed to produce and purify a fungal L-asparaginase through an aqueous two-phase micellar system (ATPMS) using Triton X-114. The fungus Penicillium sp.-encoded 2DSST1 was isolated from Cerrado soil. Plackett-Burman design followed by a 24 full factorial design was used to determine the best conditions to produce L-asparaginase. The evaluated variables were L-asparagine, L-proline, wheat bran, potato dextrose broth, ammonium sulfate, yeast extract, sucrose and glucose concentrations, incubation temperature, incubation period, and initial pH of the culture medium. L-asparaginase quantification was valued by the formation of β-aspartyl hydroxamate. The significant positive variables, L-asparagine, L-proline, potato dextrose broth, and sucrose concentrations, were evaluated at 2 levels (+ 1 and - 1) with triplicate of the central point. After 34 runs, maximum activity (2.33IU/mL) was achieved at the factorial design central point. A central composite design was performed in ATPMS at two levels (+ 1 and - 1) varying Triton X-114 concentration (w/v), separation phase temperature, and crude extract concentration (w/v). The L-asparaginase partition coefficient (K) was considered the experimental design response. Out of the 16 systems that were examined, the most promising presented a purification factor of 1.4 and a yield of 100%.

The impact of electrolyte and urea on the phase separation of Triton X-100

Abstract The temperature dependent phase separation of non-ionic surfactants is the most important characteristic of non-ionic surfactants. Herein, phase separation of Triton X-100 (TX-100) has been investigated in electrolytes and urea medium. The cloud point (CP) of TX-100 increases with the augment of the concentration of TX-100 in the electrolyte medium whereas decrease slightly with the rise of the concentration of TX-100 followed by increase with the subsequent rise of concentration in the urea medium. The CP values of TX-100 reduce in the electrolyte medium and augment in the urea medium as compared to aqueous medium. The positive values of free energy of clouding (∆Goc) signify the non-spontaneous phase separation of TX-100 in all cases. The values of enthalpy change (∆Hoc) as well as entropy change (∆Soc) of clouding were positive which reduce with the rise of TX-100 concentration and appear to be negative at the highest concentration of TX-100 employed herein in the electrolyte/urea mediums in almost all cases. The free energy of transfer (∆Goc,tr) were negative in electrolyte medium whereas positive in urea medium. The other thermodynamic properties of transfer as well as entropy-enthalpy compensation parameters were also assessed and explained with proper clarification.

A 3D Cell Death Assay to Quantitatively Determine Ferroptosis in Spheroids.

The failure of drug efficacy in clinical trials remains a big issue in cancer research. This is largely due to the limitations of two-dimensional (2D) cell cultures, the most used tool in drug screening. Nowadays, three-dimensional (3D) cultures, including spheroids, are acknowledged to be a better model of the in vivo environment, but detailed cell death assays for 3D cultures (including those for ferroptosis) are scarce. In this work, we show that a new cell death analysis method, named 3D Cell Death Assay (3DELTA), can efficiently determine different cell death types including ferroptosis and quantitatively assess cell death in tumour spheroids. Our method uses Sytox dyes as a cell death marker and Triton X-100, which efficiently permeabilizes all cells in spheroids, was used to establish 100% cell death. After optimization of Sytox concentration, Triton X-100 concentration and timing, we showed that the 3DELTA method was able to detect signals from all cells without the need to disaggregate spheroids. Moreover, in this work we demonstrated that 2D experiments cannot be extrapolated to 3D cultures as 3D cultures are less sensitive to cell death induction. In conclusion, 3DELTA is a more cost-effective way to identify and measure cell death type in 3D cultures, including spheroids.

Study of boiling behavior of surfactant added saline water over coated surface

The present article is focused on two phase heat transfer experiment, conducted over plain and porous coated circular tube of 20 mm diameter with 130 mm heating length in the quiescent pool of fluid at atmospheric pressure. The heat flux is varied from 20.19–70.22 kW/m2. The concentration of SDS and TritonX-100 are varied from 100 to 1200 ppm and 100–400 ppm in NaCl solutions respectively. It is found that higher nucleate boiling heat transfer coefficient obtained on plain tube as well as porous coated tube at SDS_400 ppm and TritonX-100_100 ppm. However, the maximum enhancement found in SDS surfactant. By increasing the concentration of SDS and TritonX-100, the nucleate boiling heat transfer decreases and minimum value found at 1200 ppm and 400 ppm respectively. As compared to the plain tube with plain NaCl solution, an enhancement of 51.43–65.74% found in the nucleate boiling heat transfer coefficient for the porous coated tube. At SDS_400 ppm and TritonX-100, an enhancement of the nucleate boiling heat transfer coefficient found about 46.42–57.71% and 36.75–50.21% for the plain tube. In general, the nucleate boiling heat transfer coefficient increases from plain NaCl to TritonX-100 to SDS for both the plain as well as porous coated surface.

Preparation of MgF2/SiO2 coating with broadband antireflective coating by using sol–gel combined with electron beam evaporation

The broadband antireflective coatings (ARCs) can effectively reduce the optical loss in the transmittance process and improve energy efficiency in solar cells. MgF2/SiO2 bilayer films were prepared on the surface of soda–lime substrates via sol–gel spin-coating combined with electron beam (e-beam) evaporation. The bottom layer was the MgF2 film obtained from e-beam evaporation, whereas the top layer was the nanoporous SiO2 film via sol–gel spin-coating method. The refractive indices of MgF2 and SiO2 layers at 550 nm are 1.38 and 1.12, respectively. The structural, morphological, and optical properties of the films were characterized. The effects of annealing temperature, Triton X-100 concentration, and spin-coating speed on the properties of the film were investigated. The transmittance spectra of the coatings were investigated, and the broadband antireflective performance of the bilayer structure was examined. A bilayer coating with optical properties close to the theoretical design results was obtained by optimizing the refractive index and the thickness of the double coatings. The solar transmittances of the soda–lime glass coated with the MgF2/SiO2 bilayer antireflective film increased by 8.89% at 400–800 nm. The multilayer antireflective film exhibits promising applications in photoelectric devices, high power laser systems, and antidazzle glasses.

Simultaneous Determination of Six Isoflavones from Puerariae Lobatae Radix by CPE-HPLC and Effect of Puerarin on Tyrosinase Activity.

Tyrosinase inhibitors with excellent inhibitory activities and lower side effects have promising applications in the fields of medicine, agriculture, food sciences and cosmetics. In this study, a method for simultaneous separation and determination of six target compounds (puerarin, daidzin, genistein, daidzein, genistin, and formononetin) in Puerariae Lobatae Radix was established by cloud point extraction (CPE) and concentration combined with high performance liquid chromatography (HPLC). To achieve high extraction yields, an ultrasound-assisted extraction method was developed based on a salt-modified Triton X-100 system. The optimal extraction conditions are: surfactant Triton X-100 concentration 0.07 g/mL, liquid-solid ratio 80:1 (mL/g), NaCl addition amount 0.6 g, equilibrium time 40 min, equilibrium temperature 70 °C. Under the optimal conditions, the total maximum extraction yield of the six target isoflavones reached 8.92 mg/g. Using l-tyrosine and l-dopa as substrates, the effects of puerarin on the monophenolase and diphenolase activity of tyrosinase activity were investigated by the enzyme kinetics method. The results showed that puerarin inhibited monophenolase activity with an IC50 of 0.537 mg/mL and activated diphenolase activity. The inhibition type of puerarin on monophenolase and the activation type of puerarin on diphenolase were analyzed by Lineweaver-Burk plots which show that puerarin showed mixed inhibition on monophenolase and mixed activation on diphenolase. Therefore, puerarin can be used as both a tyrosinase inhibitor and a tyrosinase activator.

Mass transfer from single carbon-dioxide bubbles in surfactant-electrolyte mixed aqueous solutions in vertical pipes

Mass transfer from single carbon-dioxide (CO2) bubbles rising through vertical pipes filled with surfactant−electrolyte mixed aqueous solutions was measured to study the combined effects of surfactant and electrolyte on the mass transfer. Triton X-100 and sodium chloride (NaCl) were used for surfactant and electrolyte, respectively. The surface tension, σeq, at an equilibrium state decreased with increasing the concentration of NaCl at a constant concentration of Triton X-100. The pipe diameters were 12.5 and 18.2 mm. A wide range of bubble diameter covered various bubble shapes from ellipsoidal to Taylor bubbles. The combined effects of Triton X-100 and NaCl on the Sherwood number, Sh, showed a different trend from those of alcohol and electrolyte, i.e. bubbles in the Triton X-100−NaCl mixed aqueous solution were not fully-contaminated even with the same σeq as in the condition fully-contaminated with Triton X-100, due to the difference in distributions of surfactant concentrations at the interface. The Sh of bubbles fully-contaminated with Triton X-100 were not affected by the presence of NaCl. The Sh of contaminated Taylor bubbles can be expressed as a combination of functional forms of available Sh correlations for clean and fully-contaminated Taylor bubbles.

Screening of extraction buffer concentration for protease extraction from the leaves of Syzygium polyanthum using full factorial design

This study aimed to identify the factors that significantly affect the extraction of protease from the leaves of Syzygium polyanthum. The extraction of protease from the leaves of Syzigium polyanthum was studied at different concentration of substances in the extraction buffer. A 24 full factorial experimental design was performed. The effect of four independent operating factors including; A, the concentration of potassium phosphate (KPO4); B, the concentration of Triton X-100; C, the concentration of glycerol; and D, the concentration of dithiotreitol (DTT) on the protease activity were evaluated statistically. All regression models gave a good fit to the experimental data (R2 > 99%). The maximum protease activity was detected when the extraction was performed using 3.0 M KPO4, 1% Triton X-100, 80% glycerol and 1.5 M DTT. The results revealed that the concentration of KPO4, DTT, and their interaction were statistically significant to affect the protease activity. Thus, the results obtained from this study indicate that full factorial design is useful to screen the significant factors and minimise the number of experiments.

Kinetics of Dissociation of bis(2,4,6-Tripyridyl-s-triazine)iron(II) and tris(2,2′-Bipyridyl)iron(II) in the presence of Triton X-100/Tween 80 Mixed Micellar Medium

Surface tension studies were carried on the binary surfactant mixtures over a wide range of Triton X-100 mole fractions and total surfactant concentrations to obtain critical micellar concentration values. These values were used to determine the composition of the mixed micelles and the average interaction parameter (β) which contains all the interactions of the mixed surfactants. The method is based on Rubingh′s theory using a Gauss-Newton iteration technique written in FORTRAN. The value of β was found to be -0.69 indicating synergistic behaviour i.e., combined positive catalytic effect of both the surfactants on rates of reactions. Hence, the kinetics of dissociation of bis(2,4,6-tripyridyl-striazine) iron(II) ([Fe(tptz)2]2+) and tris(2,2′-bipyridyl)iron(II) ([Fe(bipy)3]2+) were studied in the presence of Triton X-100/Tween 80 mixed micellar medium. The reactions have been carried out in the presence of mixed micelles of Triton X-100/Tween 80 at various mole fractions of Triton X-100 (αTX-100 = [Triton X-100]/([Triton X-100] + [Tween 80])) and at different total surfactant concentrations of Triton X-100 and Tween 80 (Ct = [Triton X-100] + [Tween 80]). The results show that as αTX-100 increases the rate increases for all values of Ct. Kinetic analysis has been carried out by using a simple pseudo phase model and binding constants were determined. These binding constants were found to be in agreement with the binding constants obtained spectrophotometrically.

ФЛУОРЕСЦЕНТНІ ХАРАКТЕРИСТИКИ РОЗЧИНІВ ФЛУОРЕСЦЕЇНУ ТА ЕОЗИНУ Н У ВОДНО-МІЦЕЛЯРНИХ СЕРЕДОВИЩАХ ПАР

The effect of cationic, anionic and nonionic surfactants on the fluorescence properties of fluorescein and eosin Y aqueous solutions has been investigated. It has been found that sodium dodecyl sulfate does not affect the position of the maximum wavelengths of solutions of fluorescein and eosin Y in the study of the effect of an anionic surfactant on the fluorescence emission intensity of dyes. The intensity of the signal of the dye solutions when changing the concentration of anionic surfactant changes little. As the concentration of non-ionic surfactant increases, the fluorescence emission intensity of the fluorescein solutions decreases. In contrast, with increasing concentration of Triton X-100, there is an increase in the signal intensity of solutions of more hydrophobic eosin Y with subsequent access to the "plateau" at СТХ-100≥5.1·10–2 mol/L. The position of the maxima fluorescence emission wavelengths for the fluorescein solutions in the 0-1.0·10–5 mol/L range of concentrations of cationic surfactant cetylpyridinium chloride remain unchanged. The position of the maxima shifts to the long-wavelength region of the spectrum at higher concentrations of CPC. The nature of the position dependence of the fluorescence emission maxima of eosin Y solutions in the presence of cationic surfactants is similar. The effect of fluorescence quenching has also been shown in the study of the influence of organic substances of cationic nature on the signal intensity of fluorescein solutions. It has been shown that the concentration dependence is linear in the (0.1–4.0)·10–1 mol/L range of isoniazid molecule concentrations. The data obtained can be implemented in the development of conditions and methods for the determination of pharmacologically active substances of cationic nature by reaction with fluorescein in medicines.

Optimisation of Acetylcholinesterase Extraction from the Brain of Clarias batrachus using Response Surface Methodology

Acetylcholinesterase is an important enzyme in biosensing works. It is utilized to detect insecticides and cheaper sources of the enzyme is constantly being sought. Optimisation of the acetylcholinesterase extraction from the brain of Clarias batrachus using Response Surface Methodology is reported for the first time in this study. AChE from the brain of this fish was extracted and partially purified using ammonium sulphate. The optimum range of ammonium sulphate cut was between 40 and 50%. RSM was conducted using Box-Behnken design and the optimisation value for the extraction was determined for each variable at the end of the study. The optimum concentrations of Triton X-100, PMSF, phosphate buffer and the pH for AChE extraction based on RSM were 0.75%, 1.00 mM, 0.075 M and 7.5, respectively.