The interaction of bromothymol blue(BB) with human serum albumin(HSA) was studied by electrochemical techniques and a sensitive method for proteins assay was developed. When BB interacted with HSA, the voltammetri...The interaction of bromothymol blue(BB) with human serum albumin(HSA) was studied by electrochemical techniques and a sensitive method for proteins assay was developed. When BB interacted with HSA, the voltammetric peak current value of BB decreased linearly with the concentration of HSA in a range of 1.0--40.0 mg/L, and the peak potential shifted negatively. Based on the results, a sensitive assay method for proteins, such as HSA, bovine serum albumin(BSA), and egg albumin etc. was established. This method was further applied to determining the HSA in healthy human blood samples, and the results are not significantly different from those obtained by the classic Coomassie Brilliant Blue G-250 spectrophotometic method. The detecting conditions of this method were optimized and the interaction mechanism was discussed. The results show that the electrochemical parameters(formal potential E^0, standard rate constant of the electrode reaction ks, parameter of kinetic nα) of BB have no obvious changes before and after the interaction, which indicate that BB can interact with HSA, forming an electrochemical non-active complex. The equilibrium constant(βs) and the binding ratio(m) for this complex were calculated. The m is 4 and βs is 1.41 × 10^19. This method is fast, simple, highly sensitive, and has good selectivity, which can be used in clinical measurements.展开更多
The utility of microorganisms for solubilizing the unavailable forms of potassium(K) from soil has led to renewed interest in fabrication of rapid and sensitive plate assays for their isolation and screening. The pres...The utility of microorganisms for solubilizing the unavailable forms of potassium(K) from soil has led to renewed interest in fabrication of rapid and sensitive plate assays for their isolation and screening. The present study developed a modified plate assay and compared it with previously reported methods for the isolation and screening of K-solubilizing bacteria. The newly developed plate assay is based on improved visualization of halo zone formation around the colonies on agar plates, through inclusion of an acid-base indicator dye, bromothymol blue(BTB), to modify the previously reported Aleksandrov medium. The halo zone exhibited a significant correlation(R = 0.939) with K released in liquid medium. The visualization of potential K solubilizers was improved using this method, which would help in detection of weak/non-acid producers based on secretion of organic acids in the medium. Organic acids in plate diffuse radially and form halo zones in response to reaction with the acid-base indicator dye BTB. Furthermore, K solubilization on plates with this method can be observed within 48–72 h, against the incubation time of 4–5 d needed in the earlier method. Therefore, the newly developed protocol for the plate assay was time saving, more sensitive, and beneficial in comparison to the previously reported Aleksandrov plate assay.展开更多
Two chemical methods which are commonly used for rice grain freshness determination were investigated for their efficiencies. Method 1 is made of bromothymol blue indicator, and the principle is based on indicator's ...Two chemical methods which are commonly used for rice grain freshness determination were investigated for their efficiencies. Method 1 is made of bromothymol blue indicator, and the principle is based on indicator's color which is changed in according to pH of the stored rice grains. Method 2 is based on peroxidase activity which deteriorates during storage of rice grains. Both methods were used for determination of fresh-aged rice index of six Thai-rice cultivars, four from non-waxy rice cultivars (KDML 105, Chai Nat 1, Chai Nat 2 and Phitsanulok 2) and two from waxy rice cultivars (San-pah-tawng and RD6). Rice samples were kept in the forms of paddy and polished rice. Fresh-aged rice indices were determined using both methods every two weeks over the storage period of 24 weeks (six months). It was found that both methods were capable of detecting fresh-aged rice indices. The color of chemical solutions changed with regards to the age of rice grains and it could be detected spectrophotometrically. Rice grains which have been kept as paddy provided more consistent results. Method 1 is recommended for industrial application as it is simple, efficient and inexpensive.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.20635020 and 20375020)Doctorial Foundation of the Ministry of Education of China(No.20060426001)
文摘The interaction of bromothymol blue(BB) with human serum albumin(HSA) was studied by electrochemical techniques and a sensitive method for proteins assay was developed. When BB interacted with HSA, the voltammetric peak current value of BB decreased linearly with the concentration of HSA in a range of 1.0--40.0 mg/L, and the peak potential shifted negatively. Based on the results, a sensitive assay method for proteins, such as HSA, bovine serum albumin(BSA), and egg albumin etc. was established. This method was further applied to determining the HSA in healthy human blood samples, and the results are not significantly different from those obtained by the classic Coomassie Brilliant Blue G-250 spectrophotometic method. The detecting conditions of this method were optimized and the interaction mechanism was discussed. The results show that the electrochemical parameters(formal potential E^0, standard rate constant of the electrode reaction ks, parameter of kinetic nα) of BB have no obvious changes before and after the interaction, which indicate that BB can interact with HSA, forming an electrochemical non-active complex. The equilibrium constant(βs) and the binding ratio(m) for this complex were calculated. The m is 4 and βs is 1.41 × 10^19. This method is fast, simple, highly sensitive, and has good selectivity, which can be used in clinical measurements.
基金the Department of Biotechnology (DBT), IndiaICAR-Indian Agricultural Research Institute (IARI) for providing financial support
文摘The utility of microorganisms for solubilizing the unavailable forms of potassium(K) from soil has led to renewed interest in fabrication of rapid and sensitive plate assays for their isolation and screening. The present study developed a modified plate assay and compared it with previously reported methods for the isolation and screening of K-solubilizing bacteria. The newly developed plate assay is based on improved visualization of halo zone formation around the colonies on agar plates, through inclusion of an acid-base indicator dye, bromothymol blue(BTB), to modify the previously reported Aleksandrov medium. The halo zone exhibited a significant correlation(R = 0.939) with K released in liquid medium. The visualization of potential K solubilizers was improved using this method, which would help in detection of weak/non-acid producers based on secretion of organic acids in the medium. Organic acids in plate diffuse radially and form halo zones in response to reaction with the acid-base indicator dye BTB. Furthermore, K solubilization on plates with this method can be observed within 48–72 h, against the incubation time of 4–5 d needed in the earlier method. Therefore, the newly developed protocol for the plate assay was time saving, more sensitive, and beneficial in comparison to the previously reported Aleksandrov plate assay.
基金financially supported by the Thailand Research Fund(TRF)(Grant No.MRG-WI535S078)
文摘Two chemical methods which are commonly used for rice grain freshness determination were investigated for their efficiencies. Method 1 is made of bromothymol blue indicator, and the principle is based on indicator's color which is changed in according to pH of the stored rice grains. Method 2 is based on peroxidase activity which deteriorates during storage of rice grains. Both methods were used for determination of fresh-aged rice index of six Thai-rice cultivars, four from non-waxy rice cultivars (KDML 105, Chai Nat 1, Chai Nat 2 and Phitsanulok 2) and two from waxy rice cultivars (San-pah-tawng and RD6). Rice samples were kept in the forms of paddy and polished rice. Fresh-aged rice indices were determined using both methods every two weeks over the storage period of 24 weeks (six months). It was found that both methods were capable of detecting fresh-aged rice indices. The color of chemical solutions changed with regards to the age of rice grains and it could be detected spectrophotometrically. Rice grains which have been kept as paddy provided more consistent results. Method 1 is recommended for industrial application as it is simple, efficient and inexpensive.