|
Trichothecenes toxins (Trichothecenes) single-ended class by Fusarium (Fusarium) in a variety of secondary metabolites produced by fungi, very widely distributed in nature, is the most dangerous food contaminants naturally occurring very serious health hazards to humans and animals. It can not only cause human and animal acute, chronic poisoning, carcinogenic, teratogenic, mutagenic potential hazards, but also closely linked with the occurrence of certain endemic diseases. Due to Fusarium soil Xi ranks bacteria, many Chinese medicine susceptible to Fusarium infection, many roots and tubers medicinal root rot pathogens above Fusarium can produce single-ended part of toxigenic spore the the mycophenolate olefinic toxoid Therefore, it is necessary to single-ended trichothecenes toxins in traditional Chinese medicine research, establish toxin detection method to formulate reasonable limits, to ensure that Chinese medicine is safe and effective. In view of the single-ended trichothecenes the universality and dangers of toxins in the food, agricultural, domestic and foreign single-ended trichothecenes toxins establish multiple detection analysis method. At the same time, the countries have set strict limits. However, these analysis methods is limited to the analysis of food, agricultural products, as well as dietary toxins, the analysis of single-ended trichothecenes toxins in Chinese herbal medicine is still blank. Deoxynivalenol sickle bacteria enol (DON), of snow rot of Fusarium alcohol (NIV) and T-2 toxin, the three most polluted frequent single-ended trichothecenes toxins as the representative of traditional Chinese medicine in the single-ended spore mildew trichothecene toxin detection methods. The main contents and conclusions are as follows: 1, deoxynivalenol in Fusarium Chinese medicine enol gas chromatography and mass spectrometry confirmatory method. Chinese medicine samples by distilled water extraction, purification by immunoaffinity column (IAC), N-heptafluorobutyric dibutyryl imidazole derivative derivatization reagent by gas chromatography - electron capture detection (GC-ECD) was measured, and measurement results by GC-MS spectra. The minimum detection limit of the law 2pg, the linear range 0.02-5μg/mL, different Chinese medicine In add a level 100-5000μg/kg, the recovery rate of 85.5% -97.2%, and the relative standard deviation of less than 6.3%. Analysis determination of 58 traditional Chinese medicine samples, three samples detected contains of DON pollution levels 17.21-50.54μg/kg. The results show that the method is accurate, reliable, good reproducibility, and can be used in traditional Chinese medicine DON content analysis. This is the first reported about Chinese medicine in DON pollution. 2, the establishment of a gas chromatographic detection method of Chinese medicine in the T-2 toxin. Chinese medicine extracted by methanol - water (volume ratio 80:20) mixed solvent extraction, purification by immunoaffinity column (IAC), N-heptafluorobutyric dibutyryl imidazole derivative reagent derivative by gas chromatography - electron capture detector method (GC-ECD) measured, and the measurement results of GC-MS confirmatory. Orthogonal derivatization conditions (reaction time and temperature). The minimum detection limit of the law 25pg, linear range 0.1-16μg/mL, and different Chinese medicine In add a level 50-1000μg/kg, the recovery rate of 82.2% -98.6%, and the relative standard deviation of less than 7.5%. Analysis of 32 traditional Chinese medicine samples determination of T-2 toxin, contain no samples above the detection limit. The method is simple, accurate, reliable, and can be used for the analysis of T-2 toxin detection in traditional Chinese medicine. 3, bacteria enolase and deoxynivalenol sickle Chinese medicine simultaneous determination of snow Fusarium fungus enol HPLC method. Chinese medicine samples with acetonitrile - water (80:20, V / V) mixed solvent extraction by DON specialization purified by high performance liquid chromatography - ultraviolet detector simultaneous determination. The mobile phase was acetonitrile - methanol - water (5:5:90, VNN). DON and NIV the linear range 0.2-20μg/mL, and minimum detection limit of the method NIV 2ng of DON lowest detection limit of 2.5 ng of. In add a level 500-10000μg/kg, the recovery rate of the different traditional Chinese medicine NIV in 80.8-100.3% recoveries of DON 82.8-99.9%, the relative standard deviation of 0.9-9.6%. The 30 TCM samples were analyzed Determination above the detection limit, no sample containing NIV and of DON. The same way in front of the establishment of Chinese medicine in the DON GC-ECD detection methods were compared, analyzed the pros and cons of the two methods. Detected by HPLC-UV DON, its sensitivity is lower than the GC-ECD; However, the method of handling simple, low cost of detection, and can be used for traditional Chinese medicine in the snow rot Fusarium enol and simultaneous determination of deoxynivalenol Fusarium enol; two detection methods can meet the most stringent EU limits. 4, initially established a rapid qualitative detection method of Chinese medicine and Fusarium culture in single-ended trichothecenes toxins. Samples acetonitrile - water (80:20, V / V) mixed solvent extraction, column purification, and of the through Bond Elut Mvcotoxin purification by liquid chromatography - mass spectrometric detection (LC-MS) method Determination Fusarium culture was whether the sample containing NIV , DON and T-2 three single-ended Neurospora trichothecene toxins. Mobile phase gradient elution, initial water - methanol (80:20, v / V), was increased to 10 minutes after water - methanol (20:80, V / V), to maintain 10min; Mass using electrospray ionization (ESI), SCAN scan mode. Using this method, eight kinds of sickle bacteria culture samples containing the NIV of DON and T-2, three single-ended trichothecenes toxins.
|