True and false identification method for semen armeniacae amarae and peach kernel medicinal materials, decoction pieces and formula granules based on scopoletin component

Abstract

The invention provides a method for identifying authenticity of semen armeniacae amarae and peach kernel medicinal materials, decoction pieces and formula granules based on scopoletin components, which adopts a unique double-solvent system developing solvent aiming at the scopoletin components, can quickly and effectively identify the authenticity of the semen armeniacae amarae and peach kernel medicinal materials, decoction pieces and formula granules, and can save the cost and improve the accuracy of identification of the semen armeniacae amarae and peach kernel medicinal materials, decoction pieces and formula granules. The method is high in precision and sensitivity, good in stability, convenient and fast, capable of accurately identifying the peach kernels and the bitter apricot kernels and good in application prospect.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine identification technology, and in particular to a method for identifying the authenticity of bitter almond and peach kernel medicinal materials, processed slices, and formula granules based on scopolamine lactone. Background Technology

[0002] Peach kernels and bitter almonds belong to the same family of plants and have very similar medicinal properties and components. However, their functions and indications differ. Peach kernels have the effects of promoting blood circulation and removing blood stasis, moistening the intestines and relieving constipation, and relieving cough and asthma. They are used for amenorrhea and dysmenorrhea, abdominal masses, lung abscesses and intestinal abscesses, traumatic injuries, constipation due to intestinal dryness, and cough and asthma. Bitter almonds have the effects of lowering qi, relieving cough and asthma, and moistening the intestines and relieving constipation. They are used for cough and asthma, chest fullness and excessive phlegm, and constipation due to intestinal dryness. The two should not be used interchangeably. However, because bitter almonds are cheaper than peach kernels, there is a phenomenon where bitter almonds are sometimes mixed with peach kernels for sale.

[0003] Currently, the thin-layer chromatography (TLC) identification methods for peach kernels and bitter almonds in the Chinese Pharmacopoeia and local standards all use amygdalin as a reference. Amygdalin is a common component of both peach kernels and bitter almonds, and therefore cannot distinguish between the two. There are currently no published methods for identifying the authenticity of bitter almonds and peach kernels in medicinal materials, processed slices, or formulated granules based on scopolamine lactone. Summary of the Invention

[0004] This invention provides a thin-layer chromatography method for identifying peach kernels and bitter almonds. This method can quickly and effectively identify peach kernels and bitter almonds. This method can save costs and has high precision, sensitivity and stability, and can accurately identify peach kernels and bitter almonds.

[0005] This invention provides a method for identifying the authenticity of bitter almond and peach kernel medicinal materials, processed slices, and formulation granules based on scopolamine lactone, comprising the following steps:

[0006] Take scopolamine reference standard, peach kernel and bitter almond medicinal materials or decoction pieces or formula granules to prepare scopolamine reference standard solution and peach kernel and bitter almond medicinal materials or decoction pieces or formula granules.

[0007] Thin-layer chromatography was used to detect the presence of solutions of scopolamine reference standard, peach kernel and bitter almond medicinal materials or decoction pieces or formula granules.

[0008] The developing solvent used in the thin-layer chromatography identification method includes a first developing solvent and a second developing solvent. The first developing solvent is ethyl acetate-anhydrous ethanol-water-formic acid, and the second developing solvent is toluene-ethyl acetate-anhydrous ethanol-formic acid.

[0009] Preferably, the volume ratio of ethyl acetate-anhydrous ethanol-water-formic acid in the first developing solvent is (6-10):(2-4):(0.8-1.2):(0.1-0.3).

[0010] More preferably, the volume ratio of ethyl acetate-anhydrous ethanol-water-formic acid in the first developing solvent is 8:3:1:0.2.

[0011] Preferably, the volume ratio of toluene-ethyl acetate-anhydrous ethanol-formic acid in the second developing solvent is (6-10):(3-5):(0.8-1.2):(0.4-0.6).

[0012] More preferably, the volume ratio of toluene-ethyl acetate-anhydrous ethanol-formic acid in the second developing solvent is 8:4:1:0.5.

[0013] Preferably, the preparation method of the peach kernel and bitter almond medicinal material or decoction is as follows: take peach kernel and bitter almond medicinal material powder or decoction respectively, add ethanol respectively, sonicate, filter, and evaporate to dryness to obtain peach kernel and bitter almond medicinal material or decoction.

[0014] The preparation method of the peach kernel and bitter almond formula granules is as follows: take peach kernel and bitter almond formula granules, add methanol, sonicate, filter, and evaporate to dryness to obtain a peach kernel and bitter almond formula granule solution.

[0015] Preferably, the method for preparing the scopolamine reference solution is as follows: take scopolamine reference standard, add ethanol to dissolve it, and prepare the reference solution.

[0016] Preferably, the volume fraction of the ethanol is 65-75%, and the weight-to-volume ratio of the peach kernel or bitter almond powder to the ethanol is 1 / 4-6, in g / ml.

[0017] More preferably, the volume fraction of the added ethanol is 70%, and the weight-to-volume ratio of the peach kernel or bitter almond medicinal material to the first added ethanol is 1 / 5, in g / ml.

[0018] More preferably, the concentration of the scopolamine reference solution is 25 μg / ml.

[0019] Preferably, the ultrasonic power is 250W, the frequency is 40kHz, the ultrasonic time is 30 minutes, and the filtration is carried out using medium-speed double-circuit qualitative filter paper with a pore size of 30-50 micrometers.

[0020] Preferably, the thin-layer identification method is as follows: spot peach kernel and bitter almond solution onto a thin-layer plate, develop with a developing solvent, remove, air dry, and examine under ultraviolet light to see the strip-shaped spot on the thin-layer plate with a strip width of 7-9 mm and a distance of 9-11 mm between the spotting origin and the bottom edge of the thin-layer plate. After spotting, place the plate on a thin-layer heating plate at 40-50℃ and heat for 10-20 min.

[0021] More preferably, the weight-to-volume ratio of the peach kernel or bitter almond powder to the sample is 1 / 5, in g / μL, and the sample is applied in strip form on a thin-layer plate with a strip width of 8 mm. The distance between the sample origin and the bottom edge of the thin-layer plate is 10 mm. After application, the plate is heated at 45°C for 15 min.

[0022] Preferably, the wavelength of the ultraviolet light used in the thin-layer identification method is 366 nm.

[0023] The present invention has the following beneficial effects:

[0024] The thin-layer chromatography method for identifying peach kernels and bitter almonds used in this invention is for the identification of the authenticity of bitter almonds and peach kernels in medicinal materials, processed slices, and formulation granules containing scopolamine. The dual-solvent system developing agent involved in the method is also specially designed based on scopolamine, thus exhibiting good separation of scopolamine and high sensitivity. It can quickly and effectively identify peach kernels, bitter almonds in medicinal materials, processed slices, and formulation granules, and has good application prospects. Attached Figure Description

[0025] Figure 1 This is a hydrogen nuclear magnetic resonance spectrum (NMR) image showing the characteristic components that distinguish bitter almonds from peach kernels.

[0026] Figure 2 This is a carbon NMR spectrum of the characteristic components that distinguish bitter almonds from peach kernels.

[0027] Figure 3 The results are obtained by examining peach kernel and bitter almond kernel under ultraviolet light at a wavelength of 366 nm using the thin-layer chromatography identification method for peach kernel and bitter almond kernel in Example 1 of this invention. Wherein: 1: Scopolamine; 2: Bitter almond kernel reference material; 3: Peach kernel (wild peach) reference material; 4: Peach kernel (peach) reference material; 5: Bitter almond kernel (apricot); 6: Bitter almond kernel (wild apricot); 7: Bitter almond kernel (Northeast apricot); 8: Bitter almond kernel (Siberian apricot); 9-12: Bitter almond kernel; 13-14: Peach kernel (wild peach); 15-16: Peach kernel (peach); 17-20: Peach kernel.

[0028] Figure 4 The results are obtained by examining peach kernel and bitter almond kernel under ultraviolet light at a wavelength of 366 nm using the thin-layer chromatography identification method for peach kernel and bitter almond kernel in Example 1 of this invention. Wherein: 1: Scopolamine lactone; 2: Bitter almond kernel reference material; 3: Peach kernel (wild peach) reference material; 4: Peach kernel (peach) reference material; 5-6: Bitter almond kernel (Siberian apricot) formula granules; 7-8, 12: Peach kernel (peach) formula granules; 9-10: Peach kernel (wild peach) formula granules; 11, 13: Blanched peach kernel (peach) formula granules.

[0029] Figure 5Comparative Example 1 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 of this invention, but uses petroleum ether II-chloroform-acetone as the developing solvent. The results are examined under ultraviolet light at a wavelength of 366 nm. Wherein 1: bitter almond control extract; 2-3: bitter almond raw material; 4: peach kernel control extract; 5-6: peach kernel raw material.

[0030] Figure 6 Comparative Example 2 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-ethyl acetate-anhydrous ethanol (volume ratio 15:1:4) as the developing solvent. The results are examined under ultraviolet light at a wavelength of 366 nm. Wherein 1: bitter almond control extract; 2-3: bitter almond raw material; 4: peach kernel control extract; 5-6: peach kernel raw material.

[0031] Figure 7 Comparative Example 3 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-butyl acetate-methanol (volume ratio 15:1:4) as the developing solvent. The results are examined under ultraviolet light at a wavelength of 366 nm. Wherein 1: bitter almond control extract; 2-3: bitter almond raw material; 4: peach kernel control extract; 5-6: peach kernel raw material.

[0032] Figure 8 Comparative Example 4 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-isopropanol-methanol (volume ratio 15:1:4) as the developing solvent. The results are examined under ultraviolet light at a wavelength of 366 nm. Wherein 1: bitter almond control extract; 2-3: bitter almond raw material; 4: peach kernel control extract; 5-6: peach kernel raw material.

[0033] Figure 9 Comparative Example 5 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-butyl acetate-anhydrous ethanol (volume ratio 15:1:4) as the developing solvent, and examines the results under ultraviolet light at a wavelength of 366 nm. Wherein: 1: bitter almond control extract; 2-3: bitter almond raw material; 4: peach kernel control extract; 5-6: peach kernel raw material.

[0034] Figure 10 Comparative Example 6 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-ethyl acetate-methanol (volume ratio 15:3:3) as the developing solvent. The results are examined under ultraviolet light at a wavelength of 366 nm. Wherein 1: bitter almond control extract; 2-3: bitter almond raw material; 4: peach kernel control extract; 5-6: peach kernel raw material.

[0035] Figure 11Comparative Example 7 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-ethyl acetate-methanol (volume ratio 15:3:2) as the developing solvent. The results are examined under ultraviolet light at a wavelength of 366 nm. Wherein 1: bitter almond control extract; 2-3: bitter almond raw material; 4: peach kernel control extract; 5-6: peach kernel raw material.

[0036] Figure 12 Comparative Example 8 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-ethyl acetate-methanol-formic acid (volume ratio 15:2:3:1) as the developing solvent, and examines the results under a 366nm ultraviolet light. 1. Bitter almond granules; 2. Peach kernel granules; 3. Bitter almond reference material; 4. Peach kernel (peach) reference material.

[0037] Figure 13 Comparative Example 9 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-ethyl acetate-methanol-formic acid (volume ratio 15:2:2:0.5) as the developing solvent, and examines the results under ultraviolet light at a wavelength of 366 nm. The components are: 1. Bitter almond granules; 2. Peach kernel granules; 3. Bitter almond reference material; 4. Peach kernel (peach) reference material.

[0038] Figure 14 Comparative Example 10 uses the thin-layer chromatography method for identifying peach kernels and bitter almonds as described in Example 1 of this invention, but uses toluene-ethyl acetate-methanol-formic acid (volume ratio 15:1:2:1) as the developing solvent, and examines the results under ultraviolet light at a wavelength of 366 nm. The components are: 1. Bitter almond granules; 2. Peach kernel granules; 3. Bitter almond reference material; 4. Peach kernel (peach) reference material. Detailed Implementation

[0039] The embodiments and comparative examples of the present invention are described in detail below. The embodiments and comparative examples described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. Where specific techniques or conditions are not specified in the embodiments and comparative examples, they shall be performed in accordance with the techniques or conditions described in the literature in the art or according to the product instructions. Reagents or instruments used, unless otherwise specified, are all conventional products that can be obtained commercially. Percentages or proportions, unless otherwise specified, are volume percentages. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0040] Instruments and materials:

[0041] The system includes an ATS 4 fully automated thin-layer chromatography spotter (CAMAG, Switzerland), a dual-chamber thin-layer chromatography developing tank, a TLC visualizer thin-layer chromatography camera (CAMAG, Switzerland), a balance (Sartorius, Germany), and thin-layer silica gel pre-prepared plates (Merck, Germany).

[0042] Experimental reagents and chemicals:

[0043] Reagents: Toluene, ethyl acetate, methanol, and formic acid were all analytical grade (Guangzhou Chemical Reagent Factory), and water was laboratory-prepared purified water.

[0044] The method for preparing a 10% sulfuric acid ethanol solution is as follows: slowly pour 10 ml of concentrated sulfuric acid into 90 ml of ethanol solution and let it cool.

[0045] The scopolamine reference standard involved in Example 1 of this invention was sourced from Shanghai Shidande Standard Technical Service Co., Ltd., batch number 15690.

[0046] The peach kernel (peach) reference material involved in Example 1 of this invention was sourced from the China National Institutes for Food and Drug Control, batch number: 120953-202108; the peach kernel (mountain peach) reference material was sourced from the China National Institutes for Food and Drug Control, batch number: 121560-202203; and the bitter almond reference material was sourced from the China National Institutes for Food and Drug Control, batch number: 121554-201204.

[0047] The sources of peach kernels and bitter almonds involved in Embodiment 1 of this invention are:

[0048] Figure 3 The bitter almond (apricot) corresponding to position 5 in the chromatogram is from Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number KXR(X)-202412001;

[0049] Figure 3 The bitter almond (mountain apricot) corresponding to position 6 in the chromatogram is from Anhui Keyun Baicao Pharmaceutical Co., Ltd., with batch number KXR(SX)-202412001.

[0050] Figure 3 The bitter almond (Northeast apricot) corresponding to position 7 in the chromatogram was sourced from Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number KXR(DBX)-202412001.

[0051] Figure 3 The bitter almond (Siberian apricot) corresponding to position 8 in the chromatogram was sourced from Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number KXR(XBLY)-202412001.

[0052] Figure 3The bitter almond corresponding to position 9 in the chromatogram is from Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number KXR-202506001;

[0053] Figure 3 The bitter almond corresponding to position 10 in the chromatogram is from Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number KXR-202506002;

[0054] Figure 3 The bitter almond corresponding to position 11 in the chromatogram is from Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number KXR-202506003;

[0055] Figure 3 The bitter almond corresponding to position 12 in the chromatogram was sourced from Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number KXR-202506004.

[0056] Figure 3 The peach kernel (mountain peach) corresponding to position 13 in the chromatogram was sourced from Anhui Keyun Baicao Pharmaceutical Co., Ltd., with batch number TR(ST)202412001.

[0057] Figure 3 The peach kernel (mountain peach) corresponding to position 14 in the chromatogram was sourced from Anhui Keyun Baicao Pharmaceutical Co., Ltd., with batch number TR(ST)202412002.

[0058] Figure 3 The peach kernel (peach) corresponding to position 15 in the chromatogram: Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number TR(T)202412001;

[0059] Figure 3 The peach kernel (peach) corresponding to position 16 in the chromatogram: Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch number TR(T)202412002;

[0060] Figure 3 The peach kernel at position 17 in the chromatogram corresponds to H enterprise;

[0061] Figure 3 The peach kernel corresponding to position 18 in the chromatogram is: Enterprise I;

[0062] Figure 3 The peach kernel corresponding to position 19 in the chromatogram is from Enterprise J.

[0063] Figure 3 The peach kernel corresponding to position 20 in the chromatogram is K enterprise.

[0064] The source of the peach kernel and bitter almond granules involved in Example 1 of this invention:

[0065] Figure 4The bitter almond (Siberian apricot) granules corresponding to position 5 on the chromatogram are sourced from Company A. Figure 4 The bitter almond (Siberian apricot) formulation granules corresponding to position 6 on the chromatogram are sourced from Company B. Figure 4 The source of the peach kernel (peach) formulation granules corresponding to position 7 on the chromatogram: Company C; Figure 4 The source of the peach kernel (peach) formulation granules corresponding to position 8 on the chromatogram: Company D; Figure 4 The source of the peach kernel (wild peach) formulation granules corresponding to position 9 on the chromatogram is: Company E; Figure 4 The source of the peach kernel (wild peach) formulation granules corresponding to position 10 in the chromatogram is: Company E; Figure 4 The source of the roasted peach kernel (peach) granules corresponding to position 11 in the chromatogram is: Company F; Figure 4 The source of the roasted peach kernel (peach) granules corresponding to position 12 on the chromatogram is: Company F; Figure 4 The source of the roasted peach kernel (peach) formula granules corresponding to position 13 in the chromatogram is: Company F.

[0066] Example 1

[0067] This embodiment provides a thin-layer chromatography method for identifying peach kernels and bitter almonds, including the following steps:

[0068] 1. Sample preparation

[0069] Reference solution: Take an appropriate amount of scopolamine reference standard, dissolve it in 70% ethanol, and prepare a solution containing 25 μg per ml.

[0070] Reference medicinal material solution 1: Take 1g of peach kernel and bitter almond reference medicinal materials (i.e., bitter almond reference medicinal material, peach kernel (wild peach) reference medicinal material, and peach kernel (peach) reference medicinal material), add 5ml of 70% ethanol to each, sonicate (power 250W, frequency 40kHz) for 30min, and filter through medium-speed double-circle qualitative filter paper (pore size 30-50 micrometers) to prepare the reference medicinal material solution. Figure 1 (Use this sample)

[0071] Reference herb solution 2: Take 1g of peach kernel and bitter apricot kernel reference herbs (i.e., bitter apricot kernel reference herb, peach kernel (wild peach) reference herb, and peach kernel (peach) reference herb), add 50ml of water, decoct for 30 minutes, filter through medium-speed double-circle qualitative filter paper (pore size 30-50 micrometers), evaporate the filtrate to dryness, dissolve the residue in 2ml of methanol, and use as the reference herb solution. Figure 2 (Use this sample)

[0072] Medicinal material or decoction piece solution: Take 1g of peach kernel and bitter apricot kernel medicinal material or decoction piece (i.e., bitter apricot kernel (apricot), bitter apricot kernel (mountain apricot), bitter apricot kernel (Northeast apricot), bitter apricot kernel (Siberian apricot), bitter apricot kernel, peach kernel (mountain peach), peach kernel (peach), peach kernel) respectively, add 5ml of 70% ethanol, sonicate (power 250W, frequency 40kHz) for 30min, filter through medium-speed double-circle qualitative filter paper (pore size 30-50 microns) to make medicinal material or decoction piece solution.

[0073] Formula granule solution: Take appropriate amounts of peach kernel or bitter almond formula granules (i.e., bitter almond (Siberian apricot) formula granules, peach kernel (peach) formula granules, peach kernel (mountain peach) formula granules, and roasted peach kernel (peach) formula granules), grind them into a fine powder, take 0.2g, add 2mL of methanol, sonicate (power 250W, frequency 40kHz) for 30min, filter through medium-speed double-circle qualitative filter paper (pore size 30-50 microns) to make the formula granule solution.

[0074] 2. Chromatographic conditions

[0075] Thin-layer board: Pre-fabricated silicone ordinary thin-layer board (20cm×10cm; Merck)

[0076] Spotting: 2 μL of reference standard, 10 μL of reference medicinal material, medicinal material, and decoction piece solution, and 5 μL of formula granule solution. Spot the samples separately in strip form on the thin-layer plate with a strip width of 8 mm. The distance between the spotting origin and the bottom edge of the thin-layer plate is 10 mm. After spotting, place the plate on a thin-layer heating plate at 45°C for 15 min and set aside.

[0077] Solvent system and developing agent:

[0078] Ethyl acetate-anhydrous ethanol-water-formic acid (8:3:1:0.2) Expand to 4cm

[0079] Toluene-ethyl acetate-anhydrous ethanol-formic acid (8:4:1:0.5) Expand to 8cm

[0080] Development conditions: T: 24℃, RH: 79%

[0081] Inspection: After drying, observe and photograph the image under a UV lamp (366nm).

[0082] Identification: In the spectrum corresponding to the bitter almond reference material / medicinal material / processed slices / formulation granules, there is a blue characteristic spot at the position of Rf≈0.6. By collecting and eluting the component of this characteristic spot, mass spectrometry analysis was performed, followed by nuclear magnetic resonance hydrogen spectrum and nuclear magnetic resonance carbon spectrum analysis, which further confirmed that it is scopolamine lactone structure. Figure 1 and Figure 2 The identification results of the proton and carbon spectra are shown.

[0083] The chromatograms of peach kernel (as a reference herb / medicinal material / processed slices / formula granules) showed no interference from other spots at this location, indicating that the method has good specificity and applicability.

[0084] The results are as follows Figure 3-4 As shown. The Chinese Pharmacopoeia lists bitter almonds as including four types of origin: wild apricot, Siberian apricot, Northeast apricot, or apricot; and peach kernels as including two types of origin: peach and wild peach. (From...) Figure 3 It can be seen that the bitter almond reference material (band 2) and the bitter almonds from the four origins listed in the pharmacopoeia (bands 5-8) all showed spots of the same color at the corresponding positions in the chromatogram of scopolamine reference standard. The peach kernel reference materials from the two origins (bands 3-4) did not show spots of the same color at the corresponding positions in the chromatogram of scopolamine reference standard. This proves that this method can distinguish between bitter almonds and peach kernels. When the above method was used to test commercially collected bitter almond and peach kernel medicinal materials or processed slices (bands 9-20), it was found that the bitter almond samples all showed spots of the same color at the corresponding positions in the chromatogram of scopolamine reference standard. Normal peach kernel samples (bands 13-16) did not show spots of the same color at the corresponding positions on the chromatogram of scopolamine reference standard. However, some batches of peach kernel samples (bands 17-20) showed spots of the same color at the corresponding positions on the chromatogram of scopolamine reference standard, indicating that these batches of peach kernel samples were likely adulterated with bitter almonds, but the amount of adulteration varied. Sample with band 19 had a lighter scopolamine spot, indicating a smaller amount of adulteration, while sample with band 20 had a darker scopolamine spot, indicating a larger amount of adulteration.

[0085] Depend on Figure 4 It can be seen that the bitter almond formula granules (bands 5-6) all showed spots of the same color at the corresponding positions on the chromatogram of scopolamine reference standard. Normal peach kernel formula granules (bands 7-10) did not show spots of the same color at the corresponding positions on the chromatogram of scopolamine reference standard. However, some batches of peach kernel formula granules (bands 11-13) were found to show spots of the same color at the corresponding positions on the chromatogram of scopolamine reference standard, proving that these batches of formula granules were likely adulterated with bitter almonds.

[0086] The following comparative examples illustrate that the applicant actually used many different developing solvents and different ratios in the experiment, not limited to the following comparative examples. The comparative examples are just a few examples.

[0087] Comparative Example 1

[0088] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference being that the developing solvent used was petroleum ether II-chloroform-acetone, with a volume ratio of 5:15:0.3. The results are as follows. Figure 3As shown. Among them, 1: bitter almond control extract (Guangzhou Coman Biotechnology Co., Ltd.: EKXR-20241201); 2-3: bitter almond medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: KXR 2409005, KXR 2409006 respectively); 4: peach kernel control extract (Guangzhou Coman Biotechnology Co., Ltd.: ETR-20241201); 5-6: peach kernel medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: TR2409008, TR2409009 respectively). The results are as follows. Figure 5 As shown in the figure, the spectrum obtained by this method shows fewer spots, and these spots are all common components of peach kernels and bitter almonds, making it impossible to distinguish between peach kernels and bitter almonds.

[0089] Comparative Example 2

[0090] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent was toluene-ethyl acetate-anhydrous ethanol, with a volume ratio of 15:1:4. The results are as follows: Figure 4 As shown. Among them, 1. Bitter almond control extract (Guangzhou Coman Biotechnology Co., Ltd.: EKXR-20241201); 2-3. Bitter almond medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: KXR 2409005, KXR 2409006 respectively); 4. Peach kernel control extract (Guangzhou Coman Biotechnology Co., Ltd.: ETR-20241201); 5-6. Peach kernel medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: TR2409008, TR2409009 respectively). The results are as follows. Figure 6 As shown in the figure, the spectrum obtained by this method shows a large number of spots. Peach kernels have spots at the RF values ​​where the characteristic spots of bitter almonds are located, which will cause interference and affect the identification of peach kernels and bitter almonds.

[0091] Comparative Example 3

[0092] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent used was toluene-butyl acetate-methanol, with a volume ratio of 15:1:4. The results are as follows: Figure 5 As shown. Among them, 1. Bitter almond control extract (Guangzhou Coman Biotechnology Co., Ltd.: EKXR-20241201); 2-3. Bitter almond medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: KXR 2409005, KXR 2409006 respectively); 4. Peach kernel control extract (Guangzhou Coman Biotechnology Co., Ltd.: ETR-20241201); 5-6. Peach kernel medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: TR2409008, TR2409009 respectively). The results are as follows. Figure 7As shown in the figure, the chromatogram produced by this method shows a large number of concentrated spots with poor separation. Peach kernels have spots at the same RF values ​​as bitter almonds, which can cause interference and affect the identification of peach kernels and bitter almonds.

[0093] Comparative Example 4

[0094] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent was toluene-isopropanol-methanol, with a volume ratio of 15:1:4. The results are as follows: Figure 6 As shown. Among them, 1. Bitter almond control extract (Guangzhou Coman Biotechnology Co., Ltd.: EKXR-20241201); 2-3. Bitter almond medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: KXR 2409005, KXR 2409006 respectively); 4. Peach kernel control extract (Guangzhou Coman Biotechnology Co., Ltd.: ETR-20241201); 5-6. Peach kernel medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: TR2409008, TR2409009 respectively). The results are as follows. Figure 8 As shown in the figure, the spectrum obtained by this method shows a large number of spots, which are concentrated in the middle, resulting in poor separation. Peach kernels have spots near the RF values ​​of the characteristic spots of bitter almonds, which can cause interference and affect the identification of peach kernels and bitter almonds.

[0095] Comparative Example 5

[0096] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent used was toluene-butyl acetate-anhydrous ethanol, with a volume ratio of 15:1:4. The results are as follows: Figure 7 As shown. Among them, 1. Bitter almond control extract (Guangzhou Coman Biotechnology Co., Ltd.: EKXR-20241201); 2-3. Bitter almond medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: KXR 2409005, KXR 2409006 respectively); 4. Peach kernel control extract (Guangzhou Coman Biotechnology Co., Ltd.: ETR-20241201); 5-6. Peach kernel medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: TR2409008, TR2409009 respectively). The results are as follows. Figure 9 As shown in the figure, the spectrum obtained by this method shows fewer spots, and the logarithmic spots overlap. The characteristic spots of bitter almonds also overlap with other spots, making it impossible to distinguish between peach kernels and bitter almonds.

[0097] Comparative Example 6

[0098] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent was toluene-ethyl acetate-methanol, with a volume ratio of 15:3:3. The results are as follows: Figure 8As shown. Among them, 1. Bitter almond control extract (Guangzhou Coman Biotechnology Co., Ltd.: EKXR-20241201); 2-3. Bitter almond medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: KXR 2409005, KXR 2409006 respectively); 4. Peach kernel control extract (Guangzhou Coman Biotechnology Co., Ltd.: ETR-20241201); 5-6. Peach kernel medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: TR2409008, TR2409009 respectively). The results are as follows. Figure 10 As shown in the figure, the spectrum obtained by this method displays a large number of spots, most of which are quite diffuse. The characteristic spots of bitter almonds also show significant diffusion, and if the spots are slightly weak, it is easy to misjudge that there are no characteristic spots. This is not conducive to the differentiation between peach kernels and bitter almonds.

[0099] Comparative Example 7

[0100] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent used was toluene-ethyl acetate-methanol, with a volume ratio of 15:3:2. The results are as follows: Figure 9 As shown. Among them, 1. Bitter almond control extract (Guangzhou Coman Biotechnology Co., Ltd.: EKXR-20241201); 2-3. Bitter almond medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: KXR 2409005, KXR 2409006 respectively); 4. Peach kernel control extract (Guangzhou Coman Biotechnology Co., Ltd.: ETR-20241201); 5-6. Peach kernel medicinal material (Anhui Keyun Baicao Pharmaceutical Co., Ltd., batch numbers: TR2409008, TR2409009 respectively). The results are as follows. Figure 11 As shown in the figure, the spectrum obtained by this method shows a large number of spots, and the characteristic spots of bitter almonds are poorly separated, easily overlapping with the spots below, which affects the identification of peach kernels and bitter almonds.

[0101] Comparative Example 8

[0102] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent used was toluene-ethyl acetate-methanol-formic acid, with a volume ratio of 15:2:3:1. The results are as follows: Figure 10 As shown. Among them: 1. Bitter almond granules (Shijiazhuang Yiling Pharmaceutical Co., Ltd.: B2401002); 2. Peach kernel formula granules (Anhui Hongfang Pharmaceutical Co., Ltd., batch number: 2209079); 3. Bitter almond reference material (China National Institutes for Food and Drug Control: 121554-201204); 4. Peach kernel (peach) reference material (China National Institutes for Food and Drug Control: batch number: 120953-202108). The results are as follows. Figure 12As shown in the figure, the spectrum obtained by this method shows a large number of spots, and the separation of the characteristic spots of bitter almonds is poor, which affects the identification of peach kernels and bitter almonds.

[0103] Comparative Example 9

[0104] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent was toluene-ethyl acetate-methanol-formic acid, with a volume ratio of 15:2:2:0.5. The results are as follows: Figure 11 As shown. Among them: 1. Bitter almond granules (Shijiazhuang Yiling Pharmaceutical Co., Ltd.: B2401002); 2. Peach kernel formula granules (Anhui Hongfang Pharmaceutical Co., Ltd., batch number: 2209079); 3. Bitter almond reference material (China National Institutes for Food and Drug Control: 121554-201204); 4. Peach kernel (peach) reference material (China National Institutes for Food and Drug Control: batch number: 120953-202108). The results are as follows. Figure 13 As shown in the figure, the spectrum obtained by this method shows a large number of spots, and the separation of the characteristic spots of bitter almonds is poor, which affects the identification of peach kernels and bitter almonds.

[0105] Comparative Example 10

[0106] The thin-layer chromatography method for identifying peach kernels and bitter almonds in Example 1 was used, with the difference that the developing solvent was toluene-ethyl acetate-methanol-formic acid, with a volume ratio of 15:1:2:1. The results are as follows: Figure 12 As shown. Among them: 1. Bitter almond granules (Company G); 2. Peach kernel formula granules (Company A); 3. Bitter almond reference material (China National Institutes for Food and Drug Control: 121554-201204); 4. Peach kernel (peach) reference material (China National Institutes for Food and Drug Control: batch numbers are 120953-202108). The results are as follows. Figure 14 As shown in the figure, the characteristic spots of bitter almonds in the chromatogram obtained by this method overlap with several other spots, making it impossible to distinguish between peach kernels and bitter almonds.

[0107] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for identifying the authenticity of bitter almond and peach kernel medicinal materials, processed slices, and formulated granules based on scopolamine lactone, characterized in that, Includes the following steps: Scopolamine reference standard, peach kernel and bitter almond medicinal materials or decoction pieces or formula granules were taken separately to prepare scopolamine reference standard solution and peach kernel and bitter almond medicinal materials or decoction pieces or formula granules. Thin-layer chromatography was used to detect the scopolamine reference solution, peach kernel and bitter almond medicinal materials or decoction pieces or formula granules. The developing solvent used in the thin-layer chromatography identification method includes a first developing solvent and a second developing solvent. The first developing solvent is ethyl acetate-anhydrous ethanol-water-formic acid, and the second developing solvent is toluene-ethyl acetate-anhydrous ethanol-formic acid.

2. The method according to claim 1, characterized in that, The volume ratio of ethyl acetate-anhydrous ethanol-water-formic acid in the first developing solvent is (6-10):(2-4):(0.8-1.2):(0.1-0.3).

3. The method according to claim 2, characterized in that, The volume ratio of ethyl acetate-anhydrous ethanol-water-formic acid in the first developing solvent is 8:3:1:0.

2.

4. The method according to claim 1, characterized in that, The volume ratio of toluene-ethyl acetate-anhydrous ethanol-formic acid in the second developing solvent is (6-10):(3-5):(0.8-1.2):(0.4-0.6).

5. The method according to claim 4, characterized in that, The volume ratio of toluene-ethyl acetate-anhydrous ethanol-formic acid in the second developing solvent is 8:4:1:0.

5.

6. The method according to claim 1, characterized in that: The preparation method of the peach kernel and bitter almond medicinal materials or decoction pieces solution is as follows: take peach kernel and bitter almond medicinal materials or decoction pieces powder respectively, add ethanol respectively, sonicate, filter, and evaporate to dryness to obtain peach kernel and bitter almond medicinal materials or decoction pieces solution. The preparation method of the peach kernel and bitter almond formula granules is as follows: take peach kernel and bitter almond formula granules, add methanol, sonicate, filter, and evaporate to dryness to obtain a peach kernel and bitter almond formula granule solution. The preparation method of the scopolamine reference solution is as follows: take scopolamine reference standard, add ethanol to dissolve it, and prepare the reference solution.

7. The method according to claim 6, characterized in that, The volume fraction of ethanol is 65-75%, and the weight-to-volume ratio of the peach kernel or bitter almond powder to ethanol is 1 / 4-6, in g / ml. Preferably, the volume fraction of the added ethanol is 70%, and the weight-volume ratio of the peach kernel or bitter almond medicinal material to the first added ethanol is 1 / 5, in g / ml. Preferably, the concentration of the scopolamine reference solution is 25 μg / ml.

8. The method according to claim 6, characterized in that, The ultrasound power is 250W, the frequency is 40kHz, and the ultrasound time is 30 minutes. The filtration uses medium-speed double-circuit qualitative filter paper, i.e., the pore size is 30-50 micrometers.

9. The method according to claim 1, characterized in that, The thin-layer identification method is as follows: spot peach kernel and bitter almond solution onto a thin-layer plate, develop with a developing solvent, remove, air dry, and examine under ultraviolet light to see the strip-shaped spot on the thin-layer plate. The strip width is 7-9 mm, and the distance between the spotting origin and the bottom edge of the thin-layer plate is 9-11 mm. After spotting, place the plate on a thin-layer heating plate at 40-50℃ and heat for 10-20 min. Preferably, the weight-to-volume ratio of the peach kernel or bitter almond powder to the sample is 1 / 5, in g / μL. The sample is applied in strip form on a thin-layer plate with a strip width of 8 mm. The distance between the sample origin and the bottom edge of the thin-layer plate is 10 mm. After application, the plate is heated at 45°C for 15 min.

10. The method according to claim 9, characterized in that, The thin-layer identification method uses an ultraviolet lamp with a wavelength of 366 nm.

Images