Objective To isolate endophytic fungi from the rare medicinal plants ephedra, detect the production of ephedrine alkaloids, and explore the optimum fermentation conditions.Methods The endophytic fungi were isolated from the herbaceous stems treated.The ephedrine alkaloids were preliminarily detected by Dragendorff's reagent.The fungi strains producing ephedrine alkaloids were screened by HPLC method with reference substance of ephedrine, pseudoephedrine and methephedrine.After the optimum carbon source and nitrogen source were determined with total ephedrine alkaloids as evaluation indicators by HPLC, the orthogonal test of L16 (45) was designed to investigate five levels of carbon source amount, nitrogen source amount, pH value, temperature, fermentation time.Results The ephedra herbaceous stems endophytic fungus-Es2 mycelium extract after fermentation contained ephedrine, pseudoephedrine and methephedrine.Es3 mycelium extract containing pseudoephedrine after fermentation.Lactose and ammonium sulfate were as the most suitable carbon source and nitrogen source of Es2 fermentation to produce ephedrine alkaloids.Es produced the highest yield of ephedra alkaloids when the carbon source 30 g/L, nitrogen source 4 g/L, pH5.0, temperature 30℃, fermented for 8 days.Conclusion The ephedrine alkaloids of industrial production could be realized by microorganism fermentation method, which has very important significance to alleviate the ephedrine market demand pressure on the environment.

A poly( GMA-EGDMA) coated SPME fiber was prepared using an in-situ polymerization by direct bonding to the surface of a polydopamine-modified stainless steel wire. Then the fiber was modified by sulfuric acid. A novel solid phase microextraction coating coupled to high performance liquid chromatography ( HPLC) method based on the as-prepared fiber was developed for the determination of four pharmaceuticals and personal care products ( PPCPs) in water samples. The influences of extraction parameters, including pH, extraction time, extraction temperature and salt addition were investigated. 3 mL water sample was extracted by the as-prepared fiber for 60 min at 30 ℃, and then desorbed with mobile phase for 30 min, respectively. Desorption solution was analyzed by HPLC-DAD ( diode array detection ) . The results indicated that the extraction yield of the fiber was good for four PPCPs. The linear correlation coefficients were>0. 997 with the linear range of 2-200 μg/L. The limits of detection (S/N=3) were 0. 5-5 μg/L with RSD (n=5) of 4. 1%-11. 9%. The recoveries of four PPCPs at spiked level of 20, 50, 100 μg/L were within the range of 70. 6%-105. 5%. The results showed that this method was easy, green, accurate and precise, and could be used to assay the four PPCPs in real water samples.