Development And Application Research Of High Voltage Pulsed Electric Field Extraction Equipment

Objective: Design a rapid,efficient,safe,and energy-saving traditional Chinese medicine extraction device using Pulsed Electric Field（PEF）technology to meet the extraction needs of clinical and production applications in traditional Chinese medicine.This device can effectively shorten extraction time,retain active components,and reduce production costs,thereby significantly promoting the development of the traditional Chinese medicine industry.The prevalent method for TCM extraction,namely boiling with water,suffers from drawbacks such as prolonged extraction times,high energy consumption,and significant degradation of heat-sensitive components.Therefore,there is an imminent need for an emerging TCM extraction technology to replace conventional boiling techniques.In recent years,Pulsed Electric Field（PEF）technology has garnered considerable attention as a novel non-thermal extraction processing technique.This study revolves around the design of an innovative TCM extraction device centered on PEF technology,providing insights into extraction principles and optimal process design.The primary goal of this groundbreaking device is to effectively reduce extraction time,preserve therapeutic components,and lower production costs.The integration of PEF technology presents a promising alternative to the traditional boiling method,addressing concerns related to extraction efficiency and damage to heat-sensitive constituents in traditional Chinese medicines.Methods: Design of TCM extraction equipment based on the Principle of Electroporation by Pulsed Electric Fields.This equipment design utilizes the principle of electroporation induced by pulsed electric fields for the extraction of traditional Chinese medicine.The device comprises a pulse power supply,a Chinese medicine extraction treatment chamber,and a circulation system.High-voltage probes and current probes are employed to monitor the voltage and current waveforms between the two electrodes of the pulse power supply.These waveforms are then displayed and stored on an oscilloscope.（1）Investigate the impact of different power sources,treatment chambers,and electrical parameters（peak voltage,pulse frequency）and withdrawal time on the efficiency of TCM extraction.Utilize fluorescence microscopy to observe the entry of PI dye into cells and assess the extent of cell membrane damage caused by PEF treatment.Employ scanning electron microscopy（SEM）to observe the damage to TCM cell membranes induced by PEF treatment.（2）Determine the influence of PEF treatment on the physicochemical properties of the TCM extract.Measure the production of ozone（O₃）and hydrogen peroxide（H₂O₂）in relation to peak voltage and pulse frequency.Evaluate the impact of PEF on the surface tension of water,determining the effect of PEF on water solubility.Compare the electrical conductivity and total dissolved solids（TDS）values between PEF-extracted liquid and traditionally boiled liquid to assess the solubility of bioactive substances.（3）Optimize the PEF extraction process for different medicinal plant parts using response surface methodology.Utilize HPLC to compare the fingerprint similarity of single herbs and herbal combinations.（4）Employ high-resolution mass spectrometry to qualitatively and quantitatively analyze the bioactive substances in the extracts of single herbs and herbal combinations.（5）Preliminarily assess the safety of PEF-extracted substances for consumption through cytotoxicity experiments and microbial content testing.Results:（1）In the design evaluation of power sources and treatment chambers,it was found that the extraction efficiency of alternating current pulsed power sources surpassed that of direct current pulsed power sources.Additionally,the needle-plate treatment chamber demonstrated higher extraction efficiency compared to the plate-plate treatment chamber.In the examination of electrical parameters,the extraction efficiency increased with higher voltage,reaching optimal results at a peak voltage of 3 k V.However,beyond 4 k V,a decline in extraction efficiency was observed.Similarly,the extraction efficiency increased with higher pulse frequency,reaching optimal results at 6 k Hz.However,a decrease in efficiency was noted beyond 6 k Hz.PEF treatment allowed PI dye to enter ginseng cells through damaged cell membranes,binding with DNA and emitting red fluorescence.With prolonged treatment time,fluorescence intensity gradually increased,confirming the membrane-disrupting effect of PEF.SEM observations revealed the presence of dense pores and severe structural damage in ginseng cell membranes after PEF treatment,indicating the disruptive impact of PEF on cell membranes,leading to the formation of substance exchange channels on the cell membrane surface.（2）Post-PEF treatment,O₃ production increased with higher peak voltage and pulse frequency.When the pulse peak voltage was set at 3 k V and the pulse frequency at 6 k Hz,O₃and H₂O₂ generation remained below safety thresholds,resulting in minimal damage to bioactive substances.O₃ and H₂O₂ had a short residual time in water,with almost undetectable levels 200 seconds after treatment cessation.PEF reduced water surface tension,enhancing the solubility of bioactive components and accelerating water infiltration into traditional Chinese medicine（TCM）,thereby increasing extraction efficiency.TDS and conductivity values of PEF-extracted liquid were higher than those of traditionally boiled liquid,indicating superior extraction efficiency of PEF for bioactive components compared to boiling.（3）After optimizing the extraction process for different parts of Chinese medicinal plants（roots and rhizomes,fruits,seeds,root bark,whole plants,and flowers）using response surface methodology,the optimal PEF extraction conditions were determined as follows: extraction voltage of 3 k V,pulse frequency of 6 k Hz,and extraction time of 60 seconds.Under these conditions,PEF demonstrated superior extraction efficiency for medicinal plant bioactive components compared to traditional boiling.（4）High-resolution mass spectrometry analysis revealed that PEF and boiling extraction compounds were highly similar in types and concentrations.However,PEF demonstrated superior extraction efficiency for thermally unstable,insoluble,and volatile components,such as organic acids,flavonoids,coumarins,saponins,and essential oils.Comparative analysis of PEF and boiling extraction for herbal combinations showed a high degree of similarity in the extraction of bioactive substances,with PEF exhibiting superior extraction efficiency for thermally unstable and insoluble components.（5）Microbial testing revealed that PEF exhibited better microbial eradication effects in TCM extracts compared to boiling,especially against heat-resistant microorganisms.PEF was effective in eliminating heat-resistant microorganisms that could not be deactivated by high temperatures.Evaluation of PEF extract toxicity on 293 T cells indicated no adverse effects on cell viability.Conclusion: Research indicates that both PEF technology and traditional water decoction methods can effectively extract the active ingredients from herbal medicines,demonstrating good extraction performance.However,PEF offers distinct advantages,including a shorter extraction period,higher efficiency,and lower energy consumption.Notably,PEF shows significantly superior results to traditional boiling in extracting heat-sensitive,poorly soluble,and volatile bioactive components.PEF directly acts on plant cell membranes and cell walls in the solution,causing their disruption and accelerating the release of bioactive substances.Post-treatment,PEF reduces the surface tension of water,enhancing the solubility of bioactive components and expediting the infiltration of water molecules into TCM.Charged ions,accelerated by the alternating pulsed electric field,play a stirring role in the solution,promoting the dissolution of bioactive substances at all four stages of TCM extraction: infiltration,dissolution,leaching,and diffusion.Furthermore,PEF has the capacity to eliminate various harmful microorganisms present in TCM without exhibiting toxicity to cells,indicating its safety and efficacy.Considering extraction efficiency,cost-effectiveness,safety,and environmental impact,PEF technology holds potential value in the modern production of TCM.It provides an efficient,fast,and environmentally friendly alternative for TCM extraction,contributing to the advancement of TCM modernization.