13C NMR detection of non-protein nitrogen substance adulteration in animal feed.

Illegal adulteration of melamine in animal feed and food has been widely studied. However, the risk of using substitute non-protein nitrogen substances still exists. In this study, we developed the 13C NMR method for the detection of non-protein nitrogen substance adulteration in animal feed. Three compounds, i.e., urea, melamine, and biuret, were used for method development. We found that the chemical shifts of the characteristic peaks in the carbon spectra of high-nitrogen adulterants were all between 150 and 170 ppm, whereas the chemical shifts of real protein peptide bonds (-CO-NH-) were between 170 and 180 ppm, demonstrating a good distinction between non-protein nitrogen and authentic protein. The method for analyzing melamine, urea, and biuret was validated. The R2 values were all above 0.99 within the calibration range of 0.05-2% (w/w). The limits of quantification of urea, melamine, and biuret were 0.0120%, 0.0660%, and 0.0806%, respectively. This method involves simple sample pretreatment and rapid detection while also providing high accuracy. All the sample information obtained by NMR detection does not require strict impurity removal. Compared with a previously reported 1H NMR method, the developed 13C NMR method does not require strict moisture removal to avoid active hydrogen exchange, and the interfering peak overlap is mitigated.

Extraction of ferulic acid from Angelica sinensis with supercritical CO2.

Extraction of pharmacologically active ingredient of ferulic acid from the root of Angelica sinensis with supercritical CO2 was investigated. The experimental results show that the extract yields were 0.87-4.06% at temperatures from 45 to 65 degrees Celsius and pressures from 30 to 50 MPa, and the maximum content of ferulic aicd in the extracts was about 0.35-0.37%, which is lower than that of 0.61-0.85% by conventional percolation methods. Ethanol was used as co-solvent in different ratios to raw materials in order to increase the content of ferulic acid in the extracts. The experimental results show that both the extract yields and the content of ferulic acid in the extracts increase greatly compared with pure CO2 extraction. When the ratio of ethanol to the raw material was 1.6, the content of ferulic acid in the extracts was 0.91-1.27%, indicating that supercritical fluid extraction (SFE) with CO2 in the presence of suitable co-solvent is superior to percolation in extracting polar ferulic acid from Angelica sinensis.