Method and system for detecting alpha-asarone extracting solution based on biosensor

Abstract

The invention relates to the technical field of traditional Chinese medicinal material detection, and particularly discloses an alpha-asarone extracting solution detection method and system based on a biosensor, and the method sequentially comprises the following steps: mixing medicinal material powder to be detected with an extraction solvent according to a certain solid-to-liquid ratio, and extracting volatile oil by adopting an ultrasonic or reflux method to obtain a crude extracting solution; volatile oil concentrated mother liquor is obtained; obtaining a standard solution; a combined-state electrode is obtained; obtaining electrochemical response data; and converting the electrochemical response data into the content of alpha-asarone in the sample according to the standard curve, and evaluating the precision, the recovery rate and the regeneration performance to obtain a final detection result. Recognition sites prepared by adopting the molecularly imprinted polymer are complementarily paired with template molecules in steric configuration, steric hindrance and key functional groups to realize specific recognition of alpha-asarone, so that false signals caused by matrix interference and non-specific adsorption are remarkably reduced; the electrochemical measurement method is extremely sensitive to electron transfer generated by surface bonding.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine material detection technology, specifically relating to a detection method and system for α-asarone extract based on biosensors. Background Technology

[0002] As an important active ingredient in the volatile oils of traditional Chinese medicine, the accurate quantitative detection of α-asarone is of great significance for the quality control and efficacy evaluation of medicinal materials. However, current detection technologies still have many limitations and cannot meet the requirements for efficient, sensitive, and specific analysis.

[0003] In the field of instrumental analysis, high-performance liquid chromatography (HPLC) is a commonly used method for detecting α-asarone, but its operation is complex and time-consuming. HPLC detection requires setting up cumbersome gradient elution programs, such as multi-stage adjustments of the methanol and potassium dihydrogen phosphate buffer ratios, with a single analysis often exceeding 60 minutes. More importantly, the volatile oils and pigments abundant in traditional Chinese medicine extracts can severely interfere with the detection signal, causing retention time drift. This necessitates complex purification processes using solid-phase extraction columns, increasing operating costs and posing a risk of target component loss. While gas chromatography-mass spectrometry (GC-MS) is widely used in volatile oil analysis, it is prone to co-elution and peak overlap when dealing with structurally similar terpene isomers, making it difficult to accurately distinguish between α-asarone and β-asarone.

[0004] In the field of biosensing technology, while traditional immunosensors possess specific recognition capabilities, antibody molecules are easily inactivated by environmental factors such as temperature and pH, and are prone to non-specific adsorption in complex biological matrices. Studies have shown that even with antifouling strategies such as bovine serum albumin (BSA) blocking or polyethylene glycol (PEG) modification, electrode surface impedance still increases significantly, leading to a decrease in detection sensitivity. Conventional electrochemical sensors, lacking specific recognition mechanisms, exhibit insufficient selectivity in multi-component systems of traditional Chinese medicine, failing to effectively distinguish α-asarone from other structural analogs, resulting in poor quantitative accuracy.

[0005] While molecular imprinting technology offers a new approach to solving selectivity issues, its application in the detection of α-asarone is still immature. Existing molecularly imprinted polymers (MIPs) are mostly prepared using common monomers and polymerization conditions, making it difficult to form recognition holes that precisely match the spatial configuration and functional groups of α-asarone. Furthermore, traditional MIP sensors exhibit weak electrochemical response signals and high detection limits, failing to meet the needs of detecting low-concentration samples.

[0006] In response to this problem, this application proposes a method and system for detecting α-asarone extract based on biosensors. Summary of the Invention

[0007] The purpose of this invention is to provide a method and system for detecting α-asarone extract based on biosensors, so as to solve the problems mentioned in the background art.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] One aspect of the present invention is to provide a method for constructing a biosensor, comprising the following steps:

[0010] The glassy carbon electrode or gold electrode is mechanically polished and then ultrasonically cleaned with deionized water and ethanol alternately to remove surface impurities and obtain a clean electrode surface.

[0011] Preparation of template solution: Dissolve α-asarone in a suitable organic solvent and mix with monomer and electrolyte to obtain a prepolymerization solution containing template;

[0012] The clean electrode surface is immersed in the pre-polymerization solution and electrochemical polymerization (potential scanning or constant current polymerization) is used to generate a polymer film containing a template in situ on the electrode surface, resulting in a template-embedded polymer film electrode.

[0013] The polymer membrane electrode is eluted with an organic or acidic elution solvent to remove template molecules to form recognition holes and obtain a molecularly imprinted polymer electrode.

[0014] The molecularly imprinted polymer electrode is characterized by electrochemical or impedance analysis, and nanomaterials are adsorbed or deposited on the electrode surface as needed for signal enhancement to obtain a biosensing active electrode that can be used for detection.

[0015] Preferably, the monomer is a pyrrole or styrene monomer, and the electrolyte is 0.1M PBS or a mixed solution containing methanol, so as to form imprinted sites compatible with α-asarone during the polymerization process and obtain a pre-polymerization solution.

[0016] Preferably, the electrochemical polymerization employs cyclic voltammetric scanning or constant current / potential constant electropolymerization, with a potential range of -0.2V to +0.8V and a scan rate of 10–100 mV·s. -1 The number of cycles is 5–20, used to form a dense, template-containing polymer film on the electrode surface.

[0017] Preferably, the elution process uses a methanol:acetic acid (volume ratio 9:1) or methanol / acetonitrile mixed solvent for ultrasonic or immersion elution to form highly selective recognition holes and obtain a MIP electrode.

[0018] Another aspect of the present invention is to provide a method for detecting α-asarone extract based on a biosensor, comprising the following steps:

[0019] The powdered medicinal material to be tested is mixed with the extraction solvent at a certain solid-liquid ratio, and the volatile oil is extracted by methods such as ultrasound or reflux to obtain a crude extract.

[0020] The crude extract was concentrated by rotary evaporation or nitrogen blowing and then diluted to volume with chromatographically pure methanol to obtain a concentrated mother liquor of volatile oil.

[0021] The concentrated mother liquor of the volatile oil was filtered and serially diluted with PBS and methanol in a certain ratio to prepare a standard curve solution, thus obtaining a standard solution.

[0022] The biosensing active electrode prepared by the biosensor construction method described above is contacted with the standard solution and the target molecule is specifically bound under incubation conditions to obtain a bound electrode.

[0023] Electrochemical measurements (e.g., differential pulse voltammetry or cyclic voltammetry) are performed on the bound state electrode, and the response signal is recorded to obtain electrochemical response data;

[0024] The electrochemical response data were converted into the content of α-asarone in the sample based on the standard curve, and the precision, recovery rate and regeneration performance were evaluated to obtain the final detection results.

[0025] Preferably, the extraction uses petroleum ether or n-hexane as solvent, with a solid-liquid ratio of 1:20–1:30, ultrasonic extraction for 20–40 min or reflux extraction for 1–2 h to obtain a crude extract of volatile oil.

[0026] Preferably, the incubation conditions are room temperature incubation for 5–20 min or incubation until equilibrium is reached, and after incubation, the electrode is gently washed with PBS to remove non-specific adsorption, thereby obtaining the bound electrode.

[0027] Preferably, the electrochemical measurement uses differential pulse voltammetry (DPV) or cyclic voltammetry (CV). The DPV parameters are pulse amplitude 20–100 mV, pulse interval 20–200 ms, and scan range -0.2 V to +0.6 V, to obtain a linear response and construct a calibration curve.

[0028] A biosensor-based detection system for α-asarone extract includes:

[0029] A sample pretreatment device is used to pulverize the medicinal material to be tested and mix it with the extraction solvent at a set solid-liquid ratio to obtain a crude extract.

[0030] An extraction and concentration apparatus is used to perform ultrasonic or reflux extraction on the crude extract and to concentrate the extract by rotary evaporation or nitrogen blowing to obtain a volatile oil concentrate.

[0031] A volume-distillation and filtration apparatus is used to dilute the volatile oil concentrate to volume with chromatographically pure solvent and filter it through a filter to obtain a mother liquor for analysis.

[0032] A biosensor unit for receiving the assay mother liquor and comprising an electrode assembly modified with a molecularly imprinted polymer, the electrode assembly being used to contact the mother liquor and specifically bind α-asarone at a recognition site to obtain a bound electrode;

[0033] An electrochemical measurement unit is used to perform differential pulse voltammetry, cyclic voltammetry or impedance measurement on the bound state electrode and output the raw electrochemical response signal;

[0034] The signal processing and calculation unit is used to receive the original electrochemical response signal and convert the response signal into the α-asarone content in the sample based on the stored calibration curve;

[0035] Display, recording, and regeneration apparatus for displaying or storing the content and performing chemical or electrochemical elution on the electrode assembly to achieve electrode regeneration.

[0036] Compared with the prior art, the beneficial effects of the present invention are:

[0037] (1) The recognition sites prepared by molecularly imprinted polymers (MIPs) in this invention achieve specific recognition of α-asarone by complementary pairing with template molecules in terms of spatial configuration, steric hindrance, and key functional groups, thereby significantly reducing false signals caused by matrix interference and non-specific adsorption. Electrochemical measurement methods (such as differential pulse voltammetry) are extremely sensitive to electron transfer generated by surface bonding; when conductive nanomaterials (such as gold nanoparticles) are loaded on the sensor surface, the electrochemical signal can be amplified by increasing the effective electrode area and improving the electron transport channel, thereby theoretically improving the detection sensitivity of the method.

[0038] (2) This invention utilizes the superposition of template-induced high selectivity and electrochemical amplification effect, enabling the generation of distinguishable Faraday responses even in low concentration regions. Theoretically, this method possesses a lower detection limit and can cover a wider linear response range, facilitating reliable detection of low-content samples. Combining pretreatment steps such as volatile oil extraction, concentration, and methanol volume adjustment with MIP-electrochemical detection allows for the enrichment of target molecules and the separation of complex matrix components at the physicochemical level, thereby reducing matrix noise at the source and improving quantitative accuracy and repeatability. Attached Figure Description

[0039] Figure 1 This is a flowchart of a method for constructing a biosensor according to the present invention;

[0040] Figure 2 This is a flowchart of the detection method for α-asarone extract based on a biosensor according to the present invention. Detailed Implementation

[0041] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0042] Example 1:

[0043] Please see Figure 1 As shown,

[0044] Sensor construction

[0045] Glassy carbon electrodes that have been mechanically polished and ultrasonically cleaned are used to obtain a clean electrode surface.

[0046] Preparation of template / prepolymerization solution: Dissolve α-asarone standard in a small amount of methanol, then add 0.10M PBS (pH 7.4) and pyrrole monomer (final monomer concentration approximately 0.1M) to obtain a template-containing prepolymerization solution.

[0047] The electrode was immersed in the solution and cyclic voltammetric electropolymerization was performed (potential range -0.2V to +0.8V, scan rate 50mV·s). -1 (12 cycles) is used to prepare template-containing polymer films in situ on the electrode surface.

[0048] The template polymer membrane was ultrasonically eluted with a methanol:acetic acid (9:1, v / v) solution for 3×5 min to remove the template and obtain a hole-imprinted molecular electrode (MIP-GCE).

[0049] Hollowed molecularly imprinted electrodes were equilibrated in 0.1 M PBS and electrochemically characterized (CV, EIS) to obtain a reference electrode (MIP-GCE) that can be used for detection.

[0050] Testing process

[0051] The stock solution obtained from the common pretreatment was used to prepare gradient standards and test solutions (diluted to 0.01–0.50 mg / mL). -1 (range), to obtain standard / sample solutions.

[0052] The electrode was pre-wetted in PBS and 50 μL of standard / sample solution was added to the sensitive area and incubated at room temperature for 10 min to allow α-asarone to specifically bind to the MIP recognition site, thus obtaining the bound electrode.

[0053] After incubation, wash twice with PBS to remove non-specific adsorbates and obtain a clean bound electrode.

[0054] The clean bound electrode was placed in a solution containing 5 mM [Fe(CN)6]. 3- / 4- Differential pulse voltammetry (DPV) measurements were performed in the detection buffer (parameters: pulse amplitude 50 mV, pulse duration 50 ms, scan -0.2 to +0.6 V) to record peak current I (μA).

[0055] A calibration curve (I vs C) was constructed using a series of standard solutions. The sample peak current was then substituted into the calibration equation to obtain C_sample, thereby calculating the α-asarone content (mg·g) in the sample. -1 ), and obtain the test results.

[0056] Example 2:

[0057] Instruments and reagents:

[0058] KDM type adjustable temperature electric heating mantle (Shandong Juancheng Hualu Electric Heating Instrument Co., Ltd.); AL204 type electronic analytical balance (Mettler-Toledo Instruments (Shanghai) Co., Ltd.); GC-6890A gas chromatograph (Agilent Technologies, USA); SHB-IIIA type circulating water vacuum pump (Beijing Zhongxing Weiye Century Instrument Co., Ltd.); DHG-9140A type electric thermostatic drying oven (Shanghai Yiheng Technology Co., Ltd.).

[0059] Acorus tatarinowii (Sichuan Cangxi, batch number 210102); β-asarone reference standard (Jiangsu Yongjian Pharmaceutical Technology Co., Ltd., batch number 240601); methanol (chromatographic grade, Dikama, USA); Wahaha purified water (Hangzhou Wahaha Qili Nutritional Food Co., Ltd.); synthetic zeolite (Tianjin Fuchen Chemical Reagent Factory).

[0060] Methods and Results:

[0061] Volatile oil extraction:

[0062] Add an appropriate amount of precisely weighed Acorus tatarinowii powder to a conical flask, set the ultrasonic machine temperature to 30°C, add petroleum ether to the conical flask, set the ultrasonic time and power, filter under reduced pressure after ultrasonication, perform distillation, heat and distill out most of the petroleum ether from the filtrate, and then remove the remaining petroleum ether by evaporation until a constant mass is reached, and obtain an orange-yellow oily liquid, which is Acorus tatarinowii volatile oil.

[0063] Determination of volatile oil content:

[0064] Volatile oil extraction rate (%) = (Volume of volatile oil / Mass of Acorus tatarinowii herb) × 100%

[0065] Mass of α-asarone: mα-asarone = mstandard × Atest sample / AReference standard

[0066] α-Asaurophen content = mα-Asaurophen / mAcorus tatarinowii

[0067] Determination of α-asarone content:

[0068] This experiment used GC to determine the content of α-asarone in the volatile oil of Acorus tatarinowii. The chromatographic conditions were as follows: Dimension column (30m × 0.32mm × 0.50μm); temperature program: initial temperature 90℃, ramp to 170℃ at 5℃ / min, hold for 1 min, ramp to 190℃ at 2℃ / min, ramp to 230℃ at 10℃ / min, hold for 1 min; injection port temperature: 250℃, detector temperature: 230℃; injection volume: 1μl; split ratio: 40:1; solvent: methanol; carrier gas: nitrogen. Under the above chromatographic conditions, α-asarone achieved baseline separation from other components. α-asarone showed good linearity with peak area in the range of 0.1–1.02 mg / ml (R² = 0.9992), and all methodological investigations met the requirements.

[0069] Single-factor experiment:

[0070] Ultrasound time study:

[0071] Five portions of Acorus tatarinowii powder, each weighing 5g, were added to 16 times the amount of petroleum ether. The ultrasonic power was set to 175W, and the ultrasonic time was set to 25, 30, 35, 40, and 45 min. Extraction was performed according to the method described in section 2.1. The yield was calculated, and the α-asarone content in the volatile oil was determined. The results showed that the trends of volatile oil yield and α-asarone content were basically consistent. When the reaction time was 35 min, the volatile oil yield was approximately 4.25%, and the α-asarone content was approximately 0.096 mg / g.

[0072] Ultrasonic power assessment:

[0073] Five portions of Acorus tatarinowii powder, each weighing 5g, were added to 16 times the amount of petroleum ether. The mixture was ultrasonicated for 35 minutes at power levels of 125, 150, 175, 200, and 225 W, and extracted according to the method described in section 2.1. The yield was calculated, and the α-asarone content in the volatile oil was determined. The results showed that the yield of the volatile oil and the α-asarone content exhibited a similar trend, initially increasing and then decreasing with increasing reaction time. The highest yield of volatile oil and the highest α-asarone content were achieved at an ultrasonic power of 175 W.

[0074] Feed-to-liquid ratio analysis:

[0075] Five portions of Acorus tatarinowii powder, each weighing 5g, were extracted using ultrasonic extraction at a power of 175W and a time of 35min. The extraction was performed by adding 8, 12, 16, 20, and 24 times the volume of petroleum ether, respectively. The yield was calculated, and the α-asarone content in the volatile oil was determined. The results showed that the yield of the volatile oil and the α-asarone content exhibited a similar trend. The yield of the volatile oil reached its highest value when 20 times the volume of petroleum ether was added, and the α-asarone content also approached its maximum value.

[0076] result:

[0077] Based on the results of the single-factor experiments, the effects of ultrasonic time, ultrasonic power, and solid-liquid ratio on the extraction of volatile oil from Acorus tatarinowii were investigated using orthogonal experiments, with volatile oil yield and α-asarone content as indicators. The factor level table is shown in Table 1, the orthogonal experiment results are shown in Tables 2 and 4, and the analysis of variance tables are shown in Tables 3 and 5.

[0078] Table 1 Factor Level Table

[0079]

[0080]

[0081] Table 2. Results of the orthogonal experiment (extraction rate)

[0082]

[0083] Table 3. Analysis of Variance Table

[0084]

[0085]

[0086] Table 4. Results of orthogonal experiments (α-asarone content)

[0087]

[0088] Table 5. Analysis of Variance Table

[0089]

[0090]

[0091] Table 3 shows that factors A, B, and C did not show significant effects in the experimental results. Table 2 shows that the order of influence of each factor is C>A>B, and A3>A2>A1, B3=B1>B2, C3>C2>C1; therefore, the selected combination is A3B1C3. Table 5 shows that factor A showed a significant effect in the experimental results, while factors B and C did not show significant effects. Table 4 shows that the order of influence of each factor is A>C>B, and A3>A2>A1, B3>B2>B1, C3>C2>C1; therefore, the selected combination is A3B3C3. Taking all factors into consideration, the extraction process is determined to be A3B1C3, i.e., a material-to-liquid ratio of 1:24, an extraction power of 200W, and an extraction time of 30min.

[0092] Verification experiment:

[0093] Three 5g portions of Acorus tatarinowii powder were taken and validated using the optimal extraction process described above. The average extraction rate of volatile oil in the three tests was 3.8% (RSD = 2.01%), and the average α-asarone content in the volatile oil was 0.1421 mg / g (RSD = 2.15%). The results showed no significant difference from the optimal extraction process, therefore the process was considered stable.

[0094] in conclusion:

[0095] This experiment investigated the extraction process of ultrasonic extraction using volatile oil extraction rate and α-asarone content in volatile oil as test indicators. The experimental results showed that the optimal extraction process was a material-to-liquid ratio of 1:24, an extraction power of 200W, and an extraction time of 30 minutes.

[0096] As shown above, the recognition sites prepared using molecularly imprinted polymers (MIPs) achieve specific recognition of α-asarone through complementary pairing with the template molecule in terms of spatial configuration, steric hindrance, and key functional groups, thereby significantly reducing false signals caused by matrix interference and non-specific adsorption. Electrochemical measurement methods (such as differential pulse voltammetry) are extremely sensitive to electron transfer generated by surface bonding; when conductive nanomaterials (such as gold nanoparticles) are loaded onto the sensor surface, the electrochemical signal can be amplified by increasing the effective electrode area and improving the electron transport channels, thus theoretically improving the detection sensitivity of the method.

[0097] The combined effect of template-induced high selectivity and electrochemical amplification allows for distinguishable Faraday responses even at low concentrations, theoretically giving this method a lower detection limit and enabling it to cover a wider linear response range, facilitating reliable detection of low-concentration samples. Combining pretreatment steps such as volatile oil extraction, concentration, and methanol dilution with MIP-electrochemical detection enables the enrichment of target molecules and the separation of complex matrix components at the physicochemical level, thereby reducing matrix noise at the source and improving quantitative accuracy and reproducibility.

[0098] Example 3:

[0099] A biosensor-based detection system for α-asarone extract includes:

[0100] A sample pretreatment device is used to pulverize the medicinal material to be tested and mix it with the extraction solvent at a set solid-liquid ratio to obtain a crude extract.

[0101] An extraction and concentration apparatus is used to perform ultrasonic or reflux extraction on the crude extract and to concentrate the extract by rotary evaporation or nitrogen blowing to obtain a volatile oil concentrate.

[0102] A volume-distillation and filtration apparatus is used to dilute the volatile oil concentrate to volume with chromatographically pure solvent and filter it through a filter to obtain a mother liquor for analysis.

[0103] A biosensor unit for receiving the assay mother liquor and comprising an electrode assembly modified with a molecularly imprinted polymer, the electrode assembly being used to contact the mother liquor and specifically bind α-asarone at a recognition site to obtain a bound electrode;

[0104] An electrochemical measurement unit is used to perform differential pulse voltammetry, cyclic voltammetry or impedance measurement on the bound state electrode and output the raw electrochemical response signal;

[0105] The signal processing and calculation unit is used to receive the original electrochemical response signal and convert the response signal into the α-asarone content in the sample based on the stored calibration curve;

[0106] Display, recording, and regeneration apparatus for displaying or storing the content and performing chemical or electrochemical elution on the electrode assembly to achieve electrode regeneration.

[0107] Its beneficial effects are the same as those of the embodiments of the detection method of α-asarone extract based on biosensors, and will not be repeated here.

[0108] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for constructing a biosensor, characterized in that, Includes the following steps: The glassy carbon electrode or gold electrode is mechanically polished and then ultrasonically cleaned with deionized water and ethanol alternately to remove surface impurities and obtain a clean electrode surface. α-Asauroyl ether was dissolved in a suitable organic solvent and mixed with monomers and electrolytes to obtain a prepolymerization solution containing a template. The clean electrode surface is immersed in the pre-polymerization solution and electrochemical polymerization is used to generate a polymer film containing a template in situ on the electrode surface, resulting in a template-embedded polymer film electrode. The polymer membrane electrode is eluted with an organic or acidic elution solvent to remove template molecules to form recognition holes and obtain a molecularly imprinted polymer electrode. The molecularly imprinted polymer electrode is characterized by electrochemical or impedance analysis, and nanomaterials are adsorbed or deposited on the electrode surface as needed for signal enhancement to obtain a biosensing active electrode for detection.

2. The method for constructing a biosensor according to claim 1, characterized in that, The monomer is a pyrrole or styrene monomer, and the electrolyte is 0.1M PBS or a mixed solution containing methanol, so as to form imprinted sites compatible with α-asarone during the polymerization process and obtain a pre-polymerization solution.

3. The method for constructing a biosensor according to claim 1, characterized in that, The electrochemical polymerization was performed using cyclic voltammetric scanning or constant current / potential constant electropolymerization, with a potential range of -0.2V to +0.8V and a scan rate of 10–100 mV·s. -1 The number of cycles is 5–20, used to form a dense, template-containing polymer film on the electrode surface.

4. The method for constructing a biosensor according to claim 1, characterized in that, The elution process employs ultrasonic or immersion elution with methanol and acetic acid or a mixed solvent of methanol and acetonitrile to form highly selective recognition holes and obtain MIP electrodes.

5. A method for detecting α-asarone extract based on biosensors, characterized in that, Includes the following steps: The powdered medicinal material to be tested is mixed with the extraction solvent at a certain solid-liquid ratio, and the volatile oil is extracted by ultrasonic or reflux methods to obtain a crude extract. The crude extract was concentrated by rotary evaporation or nitrogen blowing and then diluted to volume with chromatographically pure methanol to obtain a concentrated mother liquor of volatile oil. The concentrated mother liquor of the volatile oil was filtered and serially diluted with PBS and methanol in a certain ratio to prepare a standard curve solution, thus obtaining a standard solution. The biosensing active electrode prepared by the method of constructing a biosensor according to any one of claims 1-4 is contacted with the standard solution and the target molecule is specifically bound under incubation conditions to obtain a bound electrode. Electrochemical measurements were performed on the bound state electrode and the response signal was recorded to obtain electrochemical response data; The electrochemical response data were converted into the content of α-asarone in the sample based on the standard curve, and the precision, recovery rate and regeneration performance were evaluated to obtain the final detection results.

6. The method for detecting α-asarone extract based on a biosensor according to claim 5, characterized in that, The extraction was performed using petroleum ether or n-hexane as solvent, with a solid-liquid ratio of 1:20–1:30, and ultrasonic extraction for 20–40 min or reflux extraction for 1–2 h to obtain a crude extract of volatile oil.

7. The method for detecting α-asarone extract based on a biosensor according to claim 5, characterized in that, The incubation conditions are room temperature incubation for 5–20 min or incubation until equilibrium is reached, and after incubation, the electrode is gently washed with PBS to remove non-specific adsorption, thus obtaining the bound electrode.

8. The method for detecting α-asarone extract based on a biosensor according to claim 5, characterized in that, Electrochemical measurements were performed using differential pulse voltammetry (DPV) or cyclic voltammetry (CV). The DPV parameters were pulse amplitude 20–100 mV, pulse interval 20–200 ms, and scan range -0.2 V to +0.6 V. Linear responses were obtained and calibration curves were constructed.

9. A biosensor-based detection system for α-asarone extract, characterized in that, include: A sample pretreatment device is used to pulverize the medicinal material to be tested and mix it with the extraction solvent at a set solid-liquid ratio to obtain a crude extract. An extraction and concentration apparatus is used to perform ultrasonic or reflux extraction on the crude extract and to concentrate the extract by rotary evaporation or nitrogen blowing to obtain a volatile oil concentrate. A volume-distillation and filtration apparatus is used to dilute the volatile oil concentrate to volume with chromatographically pure solvent and filter it through a filter to obtain a mother liquor for analysis. A biosensor unit for receiving the assay mother liquor and comprising an electrode assembly modified with a molecularly imprinted polymer, the electrode assembly being used to contact the mother liquor and specifically bind α-asarone at a recognition site to obtain a bound electrode; An electrochemical measurement unit is used to perform differential pulse voltammetry, cyclic voltammetry or impedance measurement on the bound state electrode and output the raw electrochemical response signal; The signal processing and calculation unit is used to receive the original electrochemical response signal and convert the response signal into the α-asarone content in the sample based on the stored calibration curve; Display, recording, and regeneration apparatus for displaying or storing the content and performing chemical or electrochemical elution on the electrode assembly to achieve electrode regeneration.

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