| Inonotus obliquus is a white rot fungus that grows in Frigid Zone. As a kind of rarity medicinal fungi, it was proved that nucleic acid has many significant functions such as anticancer, AZT, curing diabetes mellitus, improving immunity, ant-oxidation, and antivirus activity et al. Inonotus obliquus is known as a kind of valuable fungi to be researched furtherly. Its chemical components are very complex, triterpenoids of its main chemical components which has unique biologic activity, has been widely applied to medicine and health care, farm, light industry et al. So it is more and more important to extract triterpenoids from Inonotus obliquus. Classical extraction methods have some common defaults, for example, taking much time, low yield and location and so on, so it can't meet with the demand of the traditional medicine. To solve the problem, a novel high intensity pulsed electric fields (PEF) technique which is a developing non-thermal technique, is applied to extract triterpenoids from the Inonotus obliquus.In the paper, the whole extracting process was studied, embedded comparison with classical extraction methods was performed and primary analysis of components of triterpenoids was achieved. It showed that PEF technology has many unique characteristics and a vast development prospect and huge potential in extraction of traditional medicine. Major works and conclusions are as follows:1. Through the single factor experiment, the paper separately studied the effect of electric field intensity, pulse number, solid/liquid ratio on the extraction yield of triterpenoids .(1) When flow velocity was 20ml/min, pulsed number was 2 and solid/liquid ratio was 1:15, with the increase of electric field intensity, the extraction yield of triterpenoids was increasing at beginning, after electric field intensity was 60kV/cm; the extraction yield of triterpenoids was decreasing. So electric field of 60kV/cm that the extraction yield of triterpenoids was the largest was chosen to extract triterpenoids from Inonotus obliquus.(2) When flow velocity was 20ml/min, electric field intensity was 25kV/cm and solid/liquid ratio was 1:15, with the increase of pulsed number, the extraction yield of triterpenoids was increasing at beginning, after pulsed number was 10; the extraction yield of triterpenoids was decreasing. So pulsed number of 10 when the extraction yield was the largest was chosen to extract triterpenoids from Inonotus obliquus.(3) When flow velocity was 20ml/min, pulsed number was 6 and electric field intensity was 45kV/cm, with the increase of solid/liquid ratio, the extraction yield of triterpenoids was increasing, after solid/liquid ratio was 1:15, the extraction yield of triterpenoids was increasing in a small range. So considering overall factors, 1:15 of solid/liquid ratio was chosen to extract triterpenoids from Inonotus obliquus.2. Through orthogonal test based on single factor experiment, a maximum of the extraction yield was achieved when flow velocity was 20ml/min, electric field intensity was 60kV/cm, pulsed number was 10 and solid/liquid ratio was 1:15. Three factors were all notable factors, among which the electric field intensity was the most important factor, the second was pulsed number and the effect of solid/liquid ratio was the least.3. A quadratic general rotary unitized design was arranged near the best parameter which was fixed by orthogonal test, and the Mathematics model was established. The regression equation was: y = 56. 4474+0.9748x1 ?2.9822x1?1.1437x?0.6664x So, the regression equation and regression coefficient were notable and level of significance was 0.01. The best combination parameter was fixed exactly: pulse number: 10, electric field intensity: 60kV/cm, solid/liquid rate 1:16.4. Triterpenoids from Inonotus obliquus was extracted through four methods of PEF, ethanol immersing, ultrasonic and microwave on the best condition, the comparison of the extraction yields of triterpenoids were as follows: ultrasonic> PEF> microwave> ethanol immersing. The extraction field was 58.02% by ultrasonic which was largest, the extraction field was 57.21% by PEF which smaller less than 1% than ultrasonic. It was believed that PEF and ultrasonic had the same extraction effect, but extraction time of PEF was 10-8 time than that of ultrasonic, so PEF method has shortened extraction time farthest and economized cost. At the same time, the extraction field of PEF was 1.23 time than that of ethanol immersing, was 1.08 time than that of microwave, extraction time was only several microsecond. In a word, comparing with ethanol immersing, ultrasonic and microwave, PEF method was more excellent.5. Triterpenoids from Inonotus obliquus was extracted through PEF on the best condition, the extraction solution was analysed by GC-MS, it is concluded that main components of triterpenoids were betulin and lanosteol, reservation time was separately 15.71min and 45.35min.6. Considering that Area(%) of chromatogram figure and the concentration of components determined were in line, Area(%)was regarded as standard to analysis qualitatively the effect of the content of triterpenoids components under different electric field intensity, the result was as follows: Area(%)value of betulin was increasing with the increase of electric field intensity at beginning, after electric field intensity was 60kV/cm, Area(%)value was decreasing, so the content of betulin was maximum when electric field intensity was 60kV/cm. Area(%)value of lanosteol was increasing with the increase of electric field intensity, after electric field intensity was 60kV/cm, Area(%)value of lanosteol was increasing in a small range, so electric field intensity 60kV/cm was the most suitable for the extraction of lanosteol.7. Area (%) was regarded as standard to analysis qualitatively the effect of the content of triterpenoids components under different pulsed number, the result was as follows: Area (%) value of betulin was increasing with the increase of pulsed number, after pulsed number was 8, Area (%) value of betulin was increasing in a small range, so pulsed number 8 was the most suitable for the extraction of betulin. Area (%) value of lanosteol was increasing with the increase of pulsed number at beginning, after pulsed number was 10, Area (%) value was decreasing, so the content of lanosteol was maximum when pulsed number was 10.8. To show the result expediently, Area*(%)was attained by calculating when the dosage of Inonotus obliquus was converted as 1g, considering that Area*(%) and Area(%) were in line, Area*(%) was regarded as standard to analysis qualitatively the effect of the content of triterpenoids components under different solid/liquid rate, the result was as follows: Area*(%) values of betulin and lanosteol were increasing with the increase of solid/liquid rate, after solid/liquid rate was 1:15, Area*(%)values of betulin and lanosteol were increasing in a small range, so solid/liquid rate 1:15 was the most suitable for the extraction of betulin and lanosteol.In a word, conditions for PEF of triterpenoids from Inonotus obliquus have been studied. PEF has been shown to be a feasible method. Compared with the conventional extraction methods, PEF provided higher extraction yield, shorter extraction time and so on. So PEF is a developing extraction method to the extraction of traditional medicine.
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