A new method for screening alpha-glucosidase inhibitors and application to marine microorganisms

A new enzyme assay method for screening alpha-glucosidase inhibitors with rapidity and simplicity was developed. The enzyme-substituted alpha-glucosidases for this assay was glucoamylase. Samples were spotted or developed on the silica gel plate. The agar solution containing substrate was poured on the plate, and paper impregnated with enzyme was layered on the agar. After incubation, an inhibitory circle would appear around the inhibitor. By using this method, more than 200 strains of marine microorganisms were screened. Among them, three active strains were found to secrete inhibitors in the culture medium.

不同类型量表的线上调查和传统纸笔调查的比较研究

During the past 11 years, with the rapid development of the Internet, more and more psychologists began to realize and take advantage of it, which led to a growing number of psychological test administrated on the internet for data collection. But there were some controversy about the reliability and representatively of this new method. To examine the applicability of the Online Survey and how different types of scales used on the internet, we first reversed the measurement instrument, then from three different levels to investigate the equivalence of online survey and paper-and-pencil assessment, namely, sample level, scale level and item level. Both Classical Test Theory and Item Response Theory were used to analyze the invariance of different types of scale applicability on the internet. The main conclusions of this study could be drawn as follows: 1. In the sample-based study, self-select sample of the online survey was compared to the random sampled sample of paper-and-pencil assessment. The results showed there were no gender difference between them (p>0.05), but the online survey sample was characterized with high qualifications, high-income and younger features (88% of the sample with post-secondary education or above, and 71% aged 20 -29 years). There were significant differences on the scores of all scales between online survey and paper-and-pencil assessment (p<0.01). With demographic controlled, there was no significant difference on the variable of Neurotic between different surveys (p>0.05). 2. With in-group design, it was proved equivalence of the scale of BI (Attitude toward Brand Importance), BT (Attitude toword Brand Switcher), Extraversion, and Conscientiousbess in the compared study in the reliability, construct validity and average scores. 3. On the item level, the results based on the Item Response Theory analysis showed that 2PLM is appropriate for personality and attitude scales. With regard to personality scale, there emerged some items with DIF in the dimensions of Openness to the experience subscale and Agreeable subscale. However, there were no significant differences about the test function. 4. Exploring the psychometrics properties of answer formats of five-, six-, seven-, ten-points, it was showed that different measurement validity between online survey and paper-and-pencil test. It was also described the lower reliability and validity of six-point scale. In conclusion, the results support the application of personality scale online, but for attitude scale, we need to choose prudently.

Determination of aliphatic amines from soil and wastewater of a paper mill by pre-column derivatization using HPLC and tandem mass spectrometry (HPLC-MS/MS)

A pre-column derivatization method for the sensitive determination of aliphatic amines using the labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) followed by HPLC with fluorescence detection and APCI/NIS identification in positive-ion mode has been developed. The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent was replaced by the 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent, BCEOC, that could easily and quickly label amines. Derivatives were stable enough to be efficiently analyzed by HPLC and showed an intense protonated molecular ion corresponding m/z [M + H](+) with APCI/MS in positive-ion mode. The collision induced dissociation of the protonated molecular ion formed characteristic fragment ions at m/z 264.1, m/z 246.0 and m/z 218.1, corresponding to the cleavages of CH2CH2O-CO, CH2CH2-OCO, and N-CH2CH2O bonds. Studies on derivatization conditions demonstrated that excellent derivatization yields close to 100% were observed with a 3 to 4-fold molar reagent excess in acetonitrile solvent, in the presence of borate buffer (pH 9.0) at 40 degrees C for 10 min. In addition, the detection responses for BCEOC derivatives were compared with those obtained with CEOC and FMOC as labeling reagents. The ratios I-BCEOC/I-CEOC and I-BCEOC/I-FMOC were, respectively, 1.40-2.76 and 1.36-2.92 for fluorescence responses (here, I was the relative fluorescence intensity). Separation of the amine derivatives had been optimized on an Eclipse XDB-C-8 column. Detection limits calculated from an 0.10 pmol injection, at a signal-to-noise ratio of 3, were 18.65-38.82 fmol (injection volume 10 mu L for fluorescence detection. The relative standard deviations for intraday determination (n = 6) of standard amine derivatives (50 pmol) were 0.0063-0.037% for retention times and 3.36-6.93% for peak areas. The mean intra-and inter-assay precision for all amines were <5.4% and 5.8%, respectively. The recoveries of amines ranged from 96 to 113%. Excellent linear responses were observed with correlation coefficients of >0.9994. The established method provided a simple and highly sensitive technique for the quantitative analysis of trace amounts of aliphatic amines from biological and natural environmental samples.