Method for determining functional components in Acer truncatum bunge leaves and application thereof

Abstract

This invention provides a method and application for the efficient determination of functional components in Acer truncatum leaves, relating to the field of plant functional component detection technology. The method includes the following steps: (1) two ultrasonic-assisted extractions using 70% methanol, followed by merging the extracts; (2) detection using high-performance liquid chromatography-tandem mass spectrometry. This invention achieves simultaneous, accurate, rapid, and stable detection of multifunctional components in Acer truncatum leaves.

Description

Technical Field

[0001] This invention provides a method and application for determining the functional components in Acer truncatum leaves, relating to the field of plant functional component detection technology. Background Technology

[0002] Acer truncatum ( A. truncatum Acer bungeana is a deciduous tree belonging to the genus Acer, endemic to China. It is mainly distributed in Northeast and North China, extending west to Shaanxi, Sichuan, and Hubei provinces, and south to Zhejiang, Jiangxi, and Anhui provinces. The leaves of Acer bungeana are rich in flavonoids, polyphenols, chlorogenic acid, and other components, and are often researched and developed as a medicinal resource.

[0003] Acer truncatum leaves are abundant, and related studies have shown that the leaves contain rich functional components, leading to a gradual increase in related research in recent years. Wang Xingyan conducted the first systematic analysis of the chemical composition of Acer truncatum leaves, creating a low-temperature, high-efficiency process for extracting tannins, flavonoids, and chlorogenic acid. Shi Weisheng et al. compared and analyzed four extraction methods for flavonoids from Acer truncatum leaves: ethanol reflux, cold soaking, boiling, and percolation. Ethanol reflux yielded the highest content, but percolation was more economical. Hu Qingping et al. found that using cellulase to break down plant cell walls resulted in better extraction of chlorogenic acid from Acer truncatum leaves. Luo Xiang et al. investigated the variation patterns of chlorogenic acid content based on different leaf collection times, quantities, and frequency. Pang Xiaoli, through measurements of flavonoids in Acer truncatum leaves during spring, summer, and autumn, concluded that the flavonoid content in Acer truncatum leaves is generally high. Gu Ronghui preliminarily isolated and identified four compounds from *Acer truncatum* leaves using a combination of chromatographic separation and purification methods, including nuclear magnetic resonance spectroscopy, high-resolution mass spectrometry (UPLC-QTOF-MS), and literature data. Among these, kaempferol-3,7-di-O-α-L-rhamnoside and 1,2,3,4,6-penta-O-galloyl-β-D-glucose were reported for the first time in *Acer truncatum* leaves. Furthermore, the latter exhibits multiple activities, providing evidence for the traditional medicinal and edible uses of *Acer truncatum* leaves. Su Yajing et al., through HPLC analysis, first discovered compounds in *Acer truncatum* and *Acer tridentata* (…). A. buergerianum Miq.) and Golden Sand Maple ( A. paxiiThe leaves of *Acer truncatum* contain corilagin. Long et al. were the first to discover the components and differences in *Acer truncatum* leaves from different regions, and for the first time revealed the influence of environmental factors on rutin, a key functional component in the leaves. Currently, most methods for detecting functional components in *Acer truncatum* leaves suffer from problems such as detecting only single components and insufficient precision. Furthermore, existing methods cannot simultaneously achieve accurate quantitative detection of multiple core functional components such as rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin. This fails to meet the practical needs for rapid, efficient, and accurate detection of multiple components in the development of *Acer truncatum* leaf resources, thus hindering the in-depth exploration and industrial application of the medicinal and edible value of *Acer truncatum* leaves. Summary of the Invention

[0004] This invention provides a method and application for determining the functional components in Acer truncatum leaves, which solves the technical problem that the multifunctional components in Acer truncatum leaves cannot be simultaneously and accurately determined and the precision is insufficient in the prior art.

[0005] This invention is implemented as follows: including the following steps: (1) Ultrasonic-assisted extraction of 70% methanol Take the sample powder, add 70% methanol, mix well and let stand for 10 min, then perform ultrasonic and centrifugation treatments in sequence. Ultrasonic treatment conditions: power 500 W, frequency 40 kHz, ultrasonic time 30 min, temperature 50℃; centrifugation conditions: 8000 rpm, centrifugation temperature 4℃, time 15 min. After centrifugation, filter and collect the supernatant; repeat the above ultrasonic and centrifugation steps, and combine the supernatants from the two extractions; take the combined supernatant, add 70% methanol to dilute to the standard mark, and filter. (2) HPLC-MS / MS detection The product obtained in step (1) was detected by HPLC-MS / MS; The chromatographic-mass spectrometry conditions were as follows: the column was an octadecylsilane-bonded silica column with dimensions of 2.1 mm × 75 mm and a particle size of 5 μm; the mobile phase was acetonitrile-0.1% formic acid as mobile phase A and 0.1% formic acid aqueous solution as mobile phase B, with gradient elution as follows: 0 min ~ 1.5 min, 95% B; 1.5 min ~ 2.5 min, 95% ~ 60% B; 2.5 min ~ 3.0 min, 60% B; 3.0 min ~ 4.0 min, 60% ~ 10% B; 4.0 min ~ 5.5 min, 10% B; 5.5 min ~ 6.0 min, 10% ~ 60% B; 6.0 min ~ 7.0 min, 60% ~ 90% B; 7.0 min ~ 10.0 min, 90% B; the flow rate was 0.3 mL / min; the column temperature was 40℃; and the injection volume was 3 μL.

[0006] As a further preferred option, filtration is performed using a 0.22 μm microporous membrane.

[0007] The method for determining the functional components in Acer truncatum leaves has been applied in the fields of germplasm resource screening, quality control of medicinal raw materials, and development and testing of functional foods.

[0008] The beneficial effects of this application are as follows: Targeted optimization is made based on the tissue characteristics of *Acer truncatum* leaves, such as their thick waxy layer and high cellulose content, as well as the physicochemical properties of flavonoids and phenolic acids in the leaves, which are easily oxidized and soluble in polar organic solvents. A pretreatment step of shaking and mixing followed by a 10-minute settling time is added to solve the problem of solvent penetration into the leaves. Simultaneously, 70% methanol is allowed to fully wet the powder and penetrate into the intercellular spaces, laying the foundation for subsequent ultrasonic disruption and extraction, thus improving the dissolution rate of components from the source. Constant temperature ultrasonic parameters of 500W / 40kHz / 50℃ are set to balance dissolution efficiency and component stability. The 500W high-power ultrasonic technology efficiently breaks down the dense fibrous cell walls of Acer truncatum leaves, releasing internally bound functional components and solving the problem of insufficient cell wall disruption caused by low-power ultrasonic technology. The 50℃ constant-temperature ultrasonic technology, compared to room temperature, moderately raises the temperature to improve the dissolution rate of functional components, while strictly controlling the temperature at 50℃ to avoid the oxidation and decomposition of flavonoids and phenolic acids caused by high temperatures (>60℃), thus balancing dissolution efficiency and component stability. The 40KHZ fixed frequency is adapted to the ultrasonic frequency for cell wall disruption of Acer truncatum leaf tissue, avoiding localized overheating of the solvent and component degradation due to excessively high frequencies, or poor cell wall disruption effect due to excessively low frequencies.

[0009] Compared with existing detection technologies, this method achieves simultaneous, accurate, rapid, and stable detection of multiple components. The operation process is standardized, and the required instruments are conventional chromatography-mass spectrometry instruments, making it easy to promote and apply in laboratories and enterprises. It provides key detection technology support for the industrialization development of Acer truncatum medicinal and edible resources. Attached Figure Description

[0010] Figure 1 These are chromatograms of the functional components of Acer truncatum leaves. Among them, A and B are chromatograms of chlorogenic acid samples and standards, respectively; C and D are chromatograms of gallic acid samples and standards, respectively; E and F are chromatograms of kaempferol samples and standards, respectively; G and H are chromatograms of quercetin samples and standards, respectively; and I and J are chromatograms of rutin samples and standards, respectively. Figure 2 These are mass spectra of the functional components of Acer truncatum leaves. Among them, A and B are the mass spectra of chlorogenic acid samples and standards, respectively; C and D are the mass spectra of gallic acid samples and standards, respectively; E and F are the mass spectra of kaempferol samples and standards, respectively; G and H are the mass spectra of quercetin samples and standards, respectively; and I and J are the mass spectra of rutin samples and standards, respectively. Detailed Implementation Example

[0011] The experimental instruments used in this embodiment are: high performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS) (chromatograph: Thermo Fisher Scientific, model: UltiMate 3000; mass spectrometer: ABI Scientific, model: 3200 Q TRAP); ultrasonic cleaner (KO-500E model).

[0012] The test reagents involved in this embodiment are: rutin, gallic acid, chlorogenic acid, kaempferol and quercetin standards (Solepro, purity >99%); methanol (Thermo Fisher Scientific, chromatographic grade); acetonitrile (Thermo Fisher Scientific, chromatographic grade).

[0013] Materials Preparation: In mid-to-late April, 10 functional leaves were selected from the 3rd, 4th, and 5th pairs of leaves on the south-facing and north-facing standard branches of each germplasm resource tree, for a total of 20 leaves, for leaf phenotypic determination. The mixed leaves after phenotypic determination were dried and stored as research samples. Five functional components with high content in Acer truncatum leaves—rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin—were selected for detection and research. The selection of these five functional components was based on a multi-dimensional consideration of the medicinal and edible value of Acer truncatum leaves, component activity, practicality of testing, and industrialization needs. Firstly, the five components cover three core functional component categories: flavonoids (rutin, kaempferol, quercetin), phenolic acids (chlorogenic acid), and tannins (gallic acid), which can comprehensively characterize the enrichment level of functional components in the leaves. Secondly, all of them are high-value active ingredients proven by research, representing the core direction for the development of Acer truncatum for medicinal and functional food applications. Thirdly, they are present in relatively high concentrations in the leaves, making them suitable for standardized and precise testing. Fourthly, the research foundation is solid, allowing direct connection to industrialization stages such as germplasm screening and raw material quality control, reducing the cost of technology implementation.

[0014] Experimental steps: ① Reference sample solution. Accurately weigh 0.05 mg of rutin, gallic acid, chlorogenic acid, kaempferol and quercetin standards into 100 mL volumetric flasks, dissolve them in 70% methanol and dilute to the mark, shake well to obtain a standard solution containing 500 ng / mL.

[0015] ② Sample solution. Ultrasonic extraction with 70% methanol was used. Approximately 0.500 g of sample powder was accurately weighed and placed in a 10 mL centrifuge tube. 70% methanol was added, and the mixture was shaken and allowed to stand for 10 min. The tube was then subjected to ultrasonic treatment (power: 500 W, frequency: 40 kHz) for 30 min at 50℃. The centrifuge tube was then centrifuged at 8000 rpm at 4℃ for 15 min, followed by filtration to collect the supernatant. The ultrasonic and centrifugation processes were repeated, and the supernatants from both extractions were combined. 1 mL of the combined supernatant was transferred to a 10 mL centrifuge tube, diluted to the 10 mL mark with 70% methanol, and filtered through a 0.22 μm microporous membrane. This double extraction of the supernatant maximizes the extraction of the active ingredient.

[0016] The reference standard solution was diluted with 70% methanol to prepare standard solutions with a gradient of 100 ng / mL, 200 ng / mL, 500 ng / mL, 1000 ng / mL, 2000 ng / mL, and 5000 ng / mL. The reference standard and the test sample were detected by HPLC-MS / MS. The chromatographic-mass spectrometry conditions were as follows: column: Innoval ODS-2 (2.1 mm × 75 mm, 5 μm); mobile phase: gradient elution using (A) acetonitrile and 0.1% formic acid and (B) 0.1% formic acid aqueous solution.

[0017] To determine the optimal gradient elution time, elution times of 5 min, 8 min, 10 min, and 15 min were set.

[0018] Specifically, the 5-minute gradient elution program is as follows: 0.0–1.0 min, 95% B; 1.0–2.0 min, 95%–60% B; 2.0–2.5 min, 60% B; 2.5–3.5 min, 60%–10% B; 3.5–4.0 min, 10% B; 4.0–5.0 min, 10%–95% B.

[0019] 8-minute gradient elution program: 0.0–1.2 min, 95% B; 1.2–2.2 min, 95%–60% B; 2.2–2.8 min, 60% B; 2.8–4.0 min, 60%–10% B; 4.0–5.5 min, 10% B; 5.5–6.5 min, 10%–60% B; 6.5–8.0 min, 60%–95% B.

[0020] 10-minute gradient elution program: 0.0–1.5 min, 95% B; 1.5–2.5 min, 95%–60% B; 2.5–3.0 min, 60% B; 3.0–4.0 min, 60%–10% B; 4.0–5.5 min, 10% B; 5.5–6.0 min, 10%–60% B; 6.0–7.0 min, 60%–90% B; 7.0–10.0 min, 90% B.

[0021] 15-minute gradient elution program: 0.0–2.0 min, 95% B; 2.0–3.5 min, 95%–60% B; 3.5–4.5 min, 60% B; 4.5–7.0 min, 60%–10% B; 7.0–9.5 min, 10% B; 9.5–11.0 min, 10%–60% B; 11.0–13.0 min, 60%–95% B; 13.0–15.0 min, 95% B.

[0022] Four parallel comparative experiments were conducted with consistent parameters such as fixed chromatographic column, mobile phase, and flow rate. Using the resolution, peak shape, and detection efficiency of the five components as core indicators, the results showed that: at 5 min, the elution resolution was <1.2 for all components, with no significant peak separation, making accurate quantification impossible; at 8 min, the elution resolution was 1.2–1.5, nearing the critical threshold, but quercetin and kaempferol showed severe peak tailing; at 15 min, the elution resolution was >2.0 for all components, but detection efficiency was low, kaempferol showed severe tailing, and the time required for a single detection increased by 50%, making it unsuitable for batch detection needs. The 10-minute gradient elution program achieves optimal balance across multiple indicators, with separations of all five functional components exceeding 2.0 (gallic acid and chlorogenic acid Rs≈2.53, chlorogenic acid and rutin Rs≈10.94, rutin and quercetin Rs≈10.71, quercetin and kaempferol Rs=4.00). The peaks are relatively symmetrical with no tailing, exhibit high mass spectrometry response values, and the single-detection time is suitable for industrial-scale batch detection needs. This is the optimal elution solution specifically designed for the detection of five functional components in Acer truncatum leaves.

[0023] In this embodiment, the specific gradient elution was selected as follows: 0 min ~ 1.5 min, 95% B; 1.5 min ~ 2.5 min, 95% ~ 60% B; 2.5 min ~ 3.0 min, 60% B; 3.0 min ~ 4.0 min, 60% ~ 10% B; 4.0 min ~ 5.5 min, 10% B; 5.5 min ~ 6.0 min, 10% ~ 60% B; 6.0 min ~ 7.0 min, 60% ~ 90% B; 7.0 min ~ 10.0 min, 90% B); flow rate: 0.3 mL / min, column temperature: 40 ℃, injection volume: 3 μL.

[0024] The mass spectrometer ion source was an ESI source in negative ion mode, using multiple reaction detection (MRM). Ion source temperature: 550 ℃; ionization voltage: -4500 V; GS1 and GS2 ion source spray gas and auxiliary gas were 50 kPa and 55 kPa, respectively, and the curtain gas was 206.84 kPa. Ion pairs and collision energies for the five components—rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin—are shown in Table 1.

[0025] Table 1. Mass spectrometry parameters of the five functional components

[0026] See Figure 1 and Figure 2 By comparing the peak times of the tested samples with those of the functional component standards, it was determined that the leaves of *Acer truncatum* contain five functional components (all samples were LS-31). The peak times of the five standards were 6.84 min, 0.73 min, 0.92 min, 7.41 min, and 7.20 min, respectively, while the peak times of the five functional components in the leaves were 6.85 min, 0.78 min, 0.96 min, 7.18 min, and 7.16 min, respectively. The peak times of the two were roughly the same, and the reaction range was reasonable, confirming that *Acer truncatum* leaves contain these five functional components.

[0027] A standard curve was plotted with the solution concentration of the functional component on the x-axis and the peak area on the y-axis, and regression analysis was performed. The results showed that the rutin standard curve was Y = 166.42X + 3.71 × 10³, r = 0.9997, and the linear range was 192.41 ng·mL. -1 ~5103 ng·mL -1 The standard curve for gallic acid was calculated as follows: Y = 72.37X + 9.31 × 10², r = 0.9979, with a linear range of 95.24 ng·mL. -1 ~5085 ng·mL -1The chlorogenic acid standard curve was calculated as follows: Y = 9.51X - 8.04 × 10², r = 0.9999, with a linear range of 102.99 ng·mL. -1 ~4988 ng·mL -1 The standard curve for kaempferol was calculated as follows: Y = 21.42X - 1.12 × 10³, r = 0.9989, with a linear range of 102.42 ng·mL. -1 ~5119 ng·mL -1 The standard curve for quercetin was Y = 12.98X - 5.52 × 10², r = 0.9991, with a linear range of 94.85 ng·mL. -1 ~5093 ng·mL -1 .

[0028] Precision test The standard solution was injected six times consecutively under the chromatographic and mass spectrometric conditions of Example 1, and the peak area was recorded. The RSDs of the precision tests for rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin were 1.81%, 2.82%, 2.64%, 2.43%, and 1.30%, respectively.

[0029] Repeatability test Six portions of the test sample powder, each approximately 0.500 g, were weighed and prepared into test solutions according to the chromatographic and mass spectrometric conditions described in Example 1. The peak areas were recorded, and the RSDs were calculated. The RSDs for repeatability tests of rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin were 1.77%, 1.43%, 2.55%, 0.70%, and 2.26%, respectively.

[0030] Stability test Six parallel sample solutions were prepared and analyzed at 0 h, 3 h, 6 h, 12 h, and 24 h according to the chromatographic and mass spectrometric conditions and methods described in Example 1. Each sample was injected three times consecutively, and the peak area was recorded and the RSD was calculated. The RSDs for the stability tests of rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin were 1.99%, 3.89%, 3.39%, 2.22%, and 2.19%, respectively. Example 2

[0031] Recovery rate of spiking Five portions of powdered Acer truncatum leaves (clonal line JC-4) were accurately weighed, each approximately 0.05 g, for a total of five portions. 0.125 mg / mL each of rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin were then accurately added to each portion. -1 0.035 mg·mL -1 0.145 mg·mL -1 0.015 mg·mL -1 0.251 mg·mL-1 1 mL of the mixed standard was prepared according to the method of Example 1 above, and the chromatographic and mass spectrometric conditions of Example 1 were used to determine the recovery rate and its RSD.

[0032] The recovery results are shown in Table 2. The units for sample content, amount added, and measured amount in Table 2 are mg.

[0033] Table 2 Results of the spiking recovery test (n = 5)

[0034] As shown in Table 2, the recoveries of rutin, gallic acid, chlorogenic acid, kaempferol, and quercetin were 96.06%, 98.12%, 97.61%, 93.82%, and 96.28%, respectively; the RSDs were 1.30%, 1.94%, 0.83%, 2.32%, and 2.05%, respectively. The correlation coefficients were all above 0.99.

[0035] In summary, the established HPLC-MS / MS method exhibits good repeatability and stability, with a high recovery rate, and can be used to determine five functional components in Acer truncatum leaves.

[0036] The determination method in Example 1 can be applied to the screening of superior germplasm resources of Acer truncatum, the quality control of Acer truncatum medicinal raw materials, and the development of functional foods. Example 3

[0037] Screening of superior germplasm resources of Acer truncatum Application method: Leaves of Acer truncatum from different geographical sources and different clones were collected (sampling method is the same as in Example 1). The content of five components, including rutin and chlorogenic acid, was determined using this method. The total content of components and the content of a single high-value component (such as chlorogenic acid) were used as indicators to screen out excellent germplasm with enriched functional components and establish a high-quality germplasm resource bank of Acer truncatum.

[0038] Application advantages: Simultaneous detection of five components enables multi-index evaluation of germplasm screening in one go, improving screening efficiency by more than 5 times compared to traditional single-component detection. Example 4

[0039] Quality control of Acer truncatum medicinal raw materials Application method: Using Acer truncatum leaves as a medicinal raw material (to extract active ingredients such as chlorogenic acid), this method is used to determine the content of five functional components, establish content threshold standards, and eliminate raw materials with substandard component content to ensure the quality stability of the medicinal raw material.

[0040] Application advantages: The RSD of precision, repeatability and stability tests are all <4%, and the recovery rate of spiking is 93.82%~98.12%. The test results are accurate and reliable, and can be used as a standardized method for quality testing of Acer truncatum medicinal raw materials. The test takes only 10 minutes, which can realize rapid batch testing of raw materials and meet the quality control needs of industrial production. Example 5

[0041] Development and Testing of Functional Foods Based on Acer truncatum Application Method: Functional teas, lozenges, extract additives, and other food products are developed using Acer truncatum leaves as raw materials. This method is used to determine the content of five functional components in the raw materials and finished products, clarify the content ratio of active ingredients in the finished products, and provide quantitative basis for food labeling and functional efficacy verification.

[0042] Application advantages: HPLC-MS / MS combined with MRM mode provides accurate qualitative and quantitative analysis, effectively avoiding interference from other components in food, resulting in more precise detection results.

[0043] Compared with existing detection technologies, this method achieves simultaneous, accurate, rapid, and stable detection of multiple components. The operation process is standardized, and the required instruments are conventional chromatography-mass spectrometry instruments, making it easy to promote and apply in laboratories and enterprises. It provides key detection technology support for the industrialization development of Acer truncatum medicinal and edible resources.

[0044] The beneficial effects of this application are as follows: Targeted optimization is made based on the tissue characteristics of *Acer truncatum* leaves, such as their thick waxy layer and high cellulose content, as well as the physicochemical properties of flavonoids and phenolic acids in the leaves, which are easily oxidized and soluble in polar organic solvents. A pretreatment step of shaking and mixing followed by a 10-minute settling time is added to solve the problem of solvent penetration into the leaves. Simultaneously, 70% methanol is allowed to fully wet the powder and penetrate into the intercellular spaces, laying the foundation for subsequent ultrasonic disruption and extraction, thus improving the dissolution rate of components from the source. Constant temperature ultrasonic parameters of 500W / 40kHz / 50℃ are set to balance dissolution efficiency and component stability. The 500W high-power ultrasonic technology efficiently breaks down the dense fibrous cell walls of Acer truncatum leaves, releasing internally bound functional components and solving the problem of insufficient cell wall disruption caused by low-power ultrasonic technology. The 50℃ constant-temperature ultrasonic technology, compared to room temperature, moderately raises the temperature to improve the dissolution rate of functional components, while strictly controlling the temperature at 50℃ to avoid the oxidation and decomposition of flavonoids and phenolic acids caused by high temperatures (>60℃), thus balancing dissolution efficiency and component stability. The 40KHZ fixed frequency is adapted to the ultrasonic frequency for cell wall disruption of Acer truncatum leaf tissue, avoiding localized overheating of the solvent and component degradation due to excessively high frequencies, or poor cell wall disruption effect due to excessively low frequencies.

[0045] Compared with existing detection technologies, this method achieves simultaneous, accurate, rapid, and stable detection of multiple components. The operation process is standardized, and the required instruments are conventional chromatography-mass spectrometry instruments, making it easy to promote and apply in laboratories and enterprises. It provides key detection technology support for the industrialization development of Acer truncatum medicinal and edible resources.

Claims

1. A method for efficiently determining the functional components in Acer truncatum leaves, characterized in that, Including the following methods: (1) Ultrasonic-assisted extraction of 70% methanol Take the sample powder, add 70% methanol, mix well and let stand for 10 min, then perform ultrasonic and centrifugation treatments in sequence. Ultrasonic treatment conditions: power 500 W, frequency 40 kHz, ultrasonic time 30 min, temperature 50℃; centrifugation conditions: 8000 rpm, centrifugation temperature 4℃, time 15 min. After centrifugation, filter and collect the supernatant; repeat the above ultrasonic and centrifugation steps, and combine the supernatants from the two extractions; take the combined supernatant, add 70% methanol to dilute to the standard mark, and filter. (2) Detection by high performance liquid chromatography-tandem mass spectrometry The chromatographic-mass spectrometry (HPLC-MS / MS) conditions for detection were as follows: the column was an octadecylsilane-bonded silica column with dimensions of 2.1 mm × 75 mm and a particle size of 5 μm; the mobile phase consisted of acetonitrile-0.1% formic acid as mobile phase A and 0.1% formic acid aqueous solution as mobile phase B, with gradient elution for 0 min ~ 1.5 min, 95% B. 1.5 min~2.5 min, 95%~60%B; 2.5 min ~3.0 min, 60%B; 3.0 min ~4.0 min, 60%~10%B; 4.0 min ~ 5.5 min, 10% B; 5.5 min ~6.0 min, 10%~60%B; 6.0 min ~7.0 min, 60%~90%B; 7.0 min ~ 10.0 min, 90% B; flow rate: 0.3 mL / min, column temperature: 40 ℃, injection volume: 3 μL.

2. The method for efficiently determining the functional components in Acer truncatum leaves according to claim 1, characterized in that, Filtration was performed using a 0.22 μm microporous membrane.

3. The application of the method for efficiently determining the functional components in Acer truncatum leaves as described in claim 1, characterized in that, This includes germplasm resource screening, quality control of medicinal raw materials, and development and testing of functional foods.

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