Method for constructing characteristic chromatogram of white hyacinth bean or its processed products and application thereof

Abstract

The present application relates to the technical field of traditional Chinese medicine analysis, in particular to a construction method of characteristic chromatogram of Lablab or its processed products and application thereof. The construction method of characteristic chromatogram of Lablab or its processed products comprises the following steps: extracting Lablab or its processed products with an extraction solvent to obtain an extract, adding the extraction solvent to the extract, filtering, and taking the filtrate as a test sample solution; feeding the test sample solution into an ultra-high performance liquid chromatograph, wherein the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, obtaining a test sample chromatogram, and introducing the test sample chromatogram into traditional Chinese medicine chromatographic fingerprint similarity evaluation software to obtain the characteristic chromatogram of Lablab or its processed products. The characteristic chromatogram of Lablab constructed by the present application has 14 characteristic peaks, and the characteristic chromatogram of Lablab processed products has 13 characteristic peaks. Lablab and Lablab processed products can be distinguished according to the characteristic chromatogram of Lablab or the characteristic chromatogram of Lablab processed products.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine analysis, in particular to a construction method of characteristic chromatogram of Dolichos lablab L. or its processed products and application thereof, and in particular to a construction method of characteristic chromatogram of Dolichos lablab L. or its processed products, a method for identifying Dolichos lablab L. and its counterfeit products and a method for detecting Dolichos lablab L. processed products. BACKGROUND

[0002] Dolichos lablab L. is the dry mature seed of Dolichos ala L. of Leguminosae. The mature fruits are collected in autumn and winter, dried, the seeds are taken out, and then dried again. Dolichos lablab tastes sweet and has a slightly warm nature, and belongs to the spleen and stomach meridians. It has the effects of invigorating the spleen, removing dampness, harmonizing the center and relieving summer-heat. It is used to treat spleen-stomach weakness, poor appetite, diarrhea, excessive leukorrhea, summer-heat vomiting and diarrhea, chest tightness and abdominal distension. Dolichos lablab mainly contains protein, fat, polysaccharide, calcium, iron, phosphorus, vitamins and other nutrients, in addition to steroids, glycosides, trigonelline and other components.

[0003] The effects of Dolichos lablab and its processed products are different. For example, fried Dolichos lablab is a processed product of Dolichos lablab, and the effects of Dolichos lablab and fried Dolichos lablab are different. The former is used for removing dampness and clearing summer-heat, and the latter is used for astringing, invigorating the spleen and stopping diarrhea. Dolichos lablab has the effects of antibiosis, antiviral, and enhancing immune function, and is used for poor appetite, diarrhea, excessive leukorrhea, summer-heat vomiting and diarrhea, chest tightness and abdominal distension, and other spleen deficiency diseases; and fried Dolichos lablab has a better therapeutic effect on poor appetite and spleen-stomach weakness. Although the fried Dolichos lablab can be distinguished according to the property of "light yellow with burnt spots" at present, since Dolichos lablab is a food and medicine, when it is used as a meal replacement, it is often ground into powder, and it is difficult to distinguish the powder by its property, and it is easy to be confused. In addition, some Chinese patent medicines contain Dolichos lablab or fried Dolichos lablab powder. In view of the fact that there is a certain difference in efficacy, in order to ensure the safety and effectiveness of the medicine, it is necessary to establish a characteristic chromatogram of Dolichos lablab or a characteristic chromatogram of fried Dolichos lablab that can distinguish the two. SUMMARY

[0004] Based on this, the present application provides a construction method of characteristic chromatogram of Dolichos lablab L. or its processed products and application thereof.

[0005] The first aspect of the present application provides a construction method of characteristic chromatogram of Dolichos lablab L. or its processed products, and the technical scheme is as follows:

[0006] A construction method of characteristic chromatogram of Dolichos lablab L. or its processed products, comprising the following steps:

[0007] The Dolichos lablab L. or its processed products are treated by extraction with an extraction solvent to obtain an extract, the extraction solvent is added to the extract, filtered, and the filtrate is taken as a test sample solution;

[0008] The test sample solution is injected into an ultra-high performance liquid chromatograph, the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, and a chromatogram of the test sample is obtained. The chromatogram of the test sample is introduced into a traditional Chinese medicine chromatographic fingerprint similarity evaluation software to obtain a characteristic spectrum of white lentil or its processed product;

[0009] The chromatographic conditions set by the ultra-high performance liquid chromatograph include:

[0010] The mobile phase includes mobile phase A and mobile phase B, mobile phase A includes acetonitrile, mobile phase B includes acetic acid aqueous solution, and the elution procedure includes:

[0011] From 0 min to 17 min, the volume percentage of mobile phase A increased from 25% to 29%, and the volume percentage of mobile phase B decreased from 75% to 71%;

[0012] From 17 to 25 minutes, the volume percentage of mobile phase A was maintained at 29%, and the volume percentage of mobile phase B was maintained at 71%;

[0013] From 25 to 43 minutes, the volume percentage of mobile phase A increased from 29% to 41%, and the volume percentage of mobile phase B decreased from 71% to 59%;

[0014] From 43 to 46 minutes, the volume percentage of mobile phase A increased from 41% to 90%, and the volume percentage of mobile phase B decreased from 59% to 10%;

[0015] From 46 min to 49 min, the volume percentage of mobile phase A was maintained at 90%, and the volume percentage of mobile phase B was maintained at 10%.

[0016] In some embodiments, the volume concentration of acetic acid in the acetic acid aqueous solution is 0.2% to 0.6%.

[0017] In some embodiments, the chromatographic conditions further include at least one of the following conditions:

[0018] 1) The stationary phase is octadecylsilane bonded silica gel, the column length is 150 mm, the inner diameter is 3 mm, and the particle size is 1.8 μm;

[0019] 2) Flow rate of 0.40 mL to 0.80 mL per minute;

[0020] 3) Column temperature is 38°C to 42°C;

[0021] 4) The carrier gas flow rate of the evaporative light scattering detector is 2.7 L / min to 3.1 L / min;

[0022] 5) The drift tube temperature of the evaporative light scattering detector is 102°C to 110°C.

[0023] In some embodiments, the construction method satisfies at least one of the following characteristics:

[0024] 1) The extraction solvent is an ethanol aqueous solution;

[0025] 2) The extraction process includes heating and reflux treatment.

[0026] In some embodiments, the processed white lentil product is fried white lentil.

[0027] The second aspect of the present invention provides a method for distinguishing white lentils from their counterfeits, and the technical solution is as follows:

[0028] A method for distinguishing white lentils from their counterfeits comprises the following steps:

[0029] Extracting the test product with an extraction solvent to obtain a test product extract, adding the extraction solvent to the test product extract, filtering, and taking the filtrate as the test product solution;

[0030] The test solution is injected into an ultra-high performance liquid chromatograph, the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, and a chromatogram of the test product is obtained, and the chromatogram of the test product is compared with the characteristic spectrum of white lentils obtained by the above-mentioned construction method;

[0031] The chromatographic conditions set on the ultra-high performance liquid chromatograph are as described above.

[0032] In some embodiments, the construction method satisfies at least one of the following characteristics:

[0033] 1) The extraction solvent is an ethanol aqueous solution;

[0034] 2) The extraction process includes heating and reflux treatment.

[0035] In some embodiments, the fake white lentil bean is at least one of white kidney bean, lentil, white eyebrow bean and white sword bean.

[0036] The third aspect of the present invention provides a method for detecting a processed white lentil product, the technical solution of which is as follows:

[0037] A method for detecting a processed white lentil product comprises the following steps:

[0038] Extracting the processed product to be tested with an extraction solvent to obtain an extract of the processed product to be tested, adding the extraction solvent to the extract of the processed product to be tested, filtering, and taking the filtrate as a solution of the processed product to be tested;

[0039] The solution of the processed product to be tested is injected into an ultra-high performance liquid chromatograph, the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, and a chromatogram of the processed product to be tested is obtained, and the chromatogram of the processed product to be tested is compared with the characteristic spectrum of the processed white lentil obtained by the above-mentioned construction method;

[0040] The chromatographic conditions set on the ultra-high performance liquid chromatograph are as described above.

[0041] In some embodiments, the construction method satisfies at least one of the following characteristics:

[0042] 1) The extraction solvent is an ethanol aqueous solution;

[0043] 2) The extraction process includes heating and reflux treatment.

[0044] Compared with the traditional solution, the present invention has the following beneficial effects:

[0045] Different from using an ultraviolet detector, the present invention uses an evaporative light scattering detector to establish a characteristic spectrum for the glycoside components of white lentils or white lentil processed products. The constructed characteristic spectrum of white lentils has a total of 14 characteristic peaks, and the characteristic spectrum of white lentils processed products has a total of 13 characteristic peaks. The two can be distinguished based on the characteristic spectrum of white lentils or the characteristic spectrum of white lentils processed products. The invention has strong specificity, can more comprehensively reflect the overall quality of white lentils or white lentil processed products, ensures the drug safety of white lentils or white lentil processed products, and provides a reference for formulating quality standards for white lentils and white lentil processed products in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the technical solutions in the embodiments of the present invention and to provide a more complete understanding of the present invention and its beneficial effects, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present application. Those skilled in the art can also derive other drawings based on these drawings without inventive effort.

[0047] Figure 1 This is an overlay of characteristic spectra of 14 batches of white lentil medicinal materials / decoction pieces;

[0048] Figure 2 This is the comparative characteristic spectrum of white lentil medicinal materials / decoction pieces, among which peak 6 is Copteroside G and peak 11 (S) is Panax notoginseng saponin Iva;

[0049] Figure 3 This is the component identification diagram of the characteristic spectrum of white lentil;

[0050] Figure 4 This is an overlay of characteristic spectra of 14 batches of stir-fried white lentil slices;

[0051] Figure 5 This is the reference characteristic spectrum of stir-fried white lentil slices, where peak 6 is Copteroside G and peak 11 (S) is Panax notoginseng saponin Iva;

[0052] Figure 6 This is a comparison chart of the characteristic spectra of white lentils and their counterfeits;

[0053] Figure 7 This is a real-life picture of stir-fried white lentils at different temperatures;

[0054] Figure 8 This is a comparison chart of the characteristic spectra of white lentils and fried white lentils at different temperatures;

[0055] Figure 9 The following are the actual pictures of fried white lentils fried at different times;

[0056] Figure 10 This is a comparison chart of the characteristic spectra of white lentils and white lentils fried at different times;

[0057] Figure 11 The chromatogram obtained for Comparative Example 1;

[0058] Figure 12 The chromatogram obtained for Comparative Example 2;

[0059] Figure 13 This is the chromatogram obtained in Comparative Example 3. DETAILED DESCRIPTION

[0060] The present invention will be further described in detail below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to provide a more thorough and comprehensive understanding of the present disclosure.

[0061] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in this specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0062] Among white lentil preparations, the characteristic spectra of white lentil formula granules are the main ones. Currently, there are two types of provincial local standards. One is the Shanghai local standard, which has a total of 5 characteristic peaks, identifying Copteroside G, Panax notoginseng saponin IVa and 3β-[(2-O-α-L-rhamnopyranosyl-2-O-β-D-galactopyranosyl-β-Dglucuronopyranosyl)oxy]-22α,24-dihydroxyolean-12-en-28-al. However, from the characteristic spectra, it cannot distinguish between white lentil and fried white lentil formula granules, and the main parameters of the characteristic spectra are not provided. The second type is the local standards of other provinces and cities except Shanghai, which have a total of 4 to 5 characteristic peaks, identifying fenugreek alkaloids and other amino acids. The specificity is not strong enough and the reference value is not great. It also cannot distinguish between white lentil and fried white lentil formula granules.

[0063] In addition, the literature research on white hyacinth bean mainly focuses on its chemical composition, clinical application and pharmacology, and there are few literatures on the quality control research of white hyacinth bean. The 2020 edition of the Chinese Pharmacopoeia contains only the properties, microscopic identification and moisture inspection of the quality control standards for white hyacinth bean, and no qualitative and quantitative quality control indicators are included; a small number of literatures report the quantitative research of total phospholipids, total saponins, picolinic acid (amino acid component), trigonelline and ginsenoside IVa in white hyacinth bean, but the method of using a single measurement indicator to achieve quality control is not enough to reflect the characteristics of the effects of traditional Chinese medicine; there are also thin-layer chromatography techniques to characterize its amino acid and nucleoside components, but the amino acid and nucleoside components lack specificity. Considering that there are mainly counterfeit products such as white kidney beans on the market as authentic white hyacinth bean medicinal materials, which seriously affects the safety of medication. Therefore, it is necessary to establish a characteristic spectrum for the chemical composition of white hyacinth bean as a standard for its qualitative identification.

[0064] To fill the gap in characteristic spectra for distinguishing white lentils and their processed products, the first aspect of the present invention provides a method for constructing a characteristic spectra of white lentils or their processed products, comprising the following steps:

[0065] Extracting white lentil or its processed product with an extraction solvent to obtain an extract, adding the extraction solvent to the extract, filtering, and taking the filtrate as the test solution;

[0066] The test sample solution is injected into an ultra-high performance liquid chromatograph, the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, and a chromatogram of the test sample is obtained. The chromatogram of the test sample is introduced into a traditional Chinese medicine chromatographic fingerprint similarity evaluation software to obtain a characteristic spectrum of white lentil or its processed product;

[0067] The chromatographic conditions set by the ultra-high performance liquid chromatograph include:

[0068] The mobile phase includes mobile phase A and mobile phase B, mobile phase A includes acetonitrile, mobile phase B includes acetic acid aqueous solution, and the elution procedure includes:

[0069] From 0 min to 17 min, the volume percentage of mobile phase A increased from 25% to 29%, and the volume percentage of mobile phase B decreased from 75% to 71%;

[0070] From 17 to 25 minutes, the volume percentage of mobile phase A was maintained at 29%, and the volume percentage of mobile phase B was maintained at 71%;

[0071] From 25 to 43 minutes, the volume percentage of mobile phase A increased from 29% to 41%, and the volume percentage of mobile phase B decreased from 71% to 59%;

[0072] From 43 to 46 minutes, the volume percentage of mobile phase A increased from 41% to 90%, and the volume percentage of mobile phase B decreased from 59% to 10%;

[0073] From 46 min to 49 min, the volume percentage of mobile phase A was maintained at 90%, and the volume percentage of mobile phase B was maintained at 10%.

[0074] After the inventor consulted the materials, it was found that in the quality standard of “White Lentil Formula Granules” issued by Shanghai, after adding the extraction solvent to the extract, the above liquid was eluted with a solid phase extraction column to obtain a test solution. Different from the above method, the present invention directly filters the above liquid to obtain a test solution containing glycoside components. At the same time, the quality standard of “White Lentil Formula Granules” issued by Shanghai uses an ultraviolet detector, and in other characteristic spectra or content detections of white lentils or their preparations, ultraviolet detectors are also used. It can be seen that the use of ultraviolet detectors has become a trend in the characteristic spectra or content detection of white lentils or their preparations. The present invention breaks this trend and uses an evaporative light scattering detector to respond to the glycoside components in the test solution. Through the above method, the present invention establishes a characteristic spectrum for the glycoside components of white lentils or white lentil processed products. The constructed characteristic spectrum of white lentils has a total of 14 characteristic peaks, and the characteristic spectrum of white lentils processed products has a total of 13 characteristic peaks. The two can be distinguished according to the characteristic spectrum of white lentils or the characteristic spectrum of white lentils processed products. The method has strong specificity, can more comprehensively reflect the overall quality of white lentils or white lentil processed products, ensures the drug safety of white lentils or white lentil processed products, and provides a reference for formulating quality standards for white lentils and white lentil processed products in the future.

[0075] Optionally, the volume concentration of acetic acid in the acetic acid aqueous solution is 0.2% to 0.6%. For example, the volume concentration of acetic acid in the acetic acid aqueous solution is 0.2%, 0.3%, 0.4%, 0.5%, or 0.6%.

[0076] Optionally, the chromatographic conditions further include: the stationary phase is octadecylsilane bonded silica gel. In some embodiments, the chromatographic column has a length of 150 mm, an inner diameter of 3 mm, and a particle size of 1.8 μm.

[0077] Optionally, the chromatographic conditions further include: a flow rate of 0.40 mL to 0.80 mL per minute, for example, a flow rate of 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, or 0.8 mL per minute.

[0078] Optionally, the chromatographic conditions further include: a column temperature of 38° C. to 42° C. For example, the column temperature is 38° C., 39° C., 40° C., 41° C., or 42° C.

[0079] Optionally, the chromatographic conditions further include: a carrier gas flow rate of the evaporative light scattering detector of 2.7 L / min to 3.1 L / min, for example, a carrier gas flow rate of 2.7 L / min, 2.8 L / min, 2.9 L / min, 3.0 L / min, or 3.1 L / min.

[0080] Optionally, the chromatographic conditions further include: the drift tube temperature of the evaporative light scattering detector is 102° C. to 110° C. For example, the drift tube temperature is 102° C., 104° C., 106° C., 108° C., or 110° C.

[0081] Optionally, the extraction solvent is an ethanol-water solution. Further optionally, the volume concentration of ethanol in the ethanol-water solution is 60% to 80%. For example, the volume concentration of ethanol in the ethanol-water solution is 60%, 65%, 70%, 75%, or 80%.

[0082] Optionally, the extraction process includes a heating reflux process. Further, optionally, the heating reflux process lasts for 20 to 40 minutes. For example, the heating reflux process lasts for 20, 25, 30, 35, or 40 minutes. Further, optionally, the heating reflux process is performed at a temperature of 80 to 90°C. For example, the heating reflux process is performed at a temperature of 80°C, 85°C, or 90°C.

[0083] Optionally, the extraction process further comprises the following steps:

[0084] After the heating and reflux treatment, the mixture is filtered to obtain a reaction residue and an extract, and the extract is dried to obtain the extract.

[0085] Optionally, the extraction process further comprises the following steps:

[0086] After the heating and reflux treatment, the reaction residue and the extract are filtered to obtain a reaction residue and an extracting solution, the reaction residue is washed with the extraction solvent to obtain a washing solution, the extracting solution and the washing solution are mixed, and dried to obtain the extract.

[0087] Optionally, the processed white lentil product is fried white lentil. It can be understood that the fried white lentil can be fried white lentil slices.

[0088] It can be understood that the white lentil can be a white lentil medicinal material or a white lentil decoction piece.

[0089] The characteristic spectrum of white lentils constructed above can also be used to distinguish white lentils from their counterfeits, filling the gap in characteristic spectrum for distinguishing white lentils from their counterfeits. A second aspect of the present invention provides a method for distinguishing white lentils from their counterfeits, which comprises the following steps:

[0090] Extracting the test product with an extraction solvent to obtain a test product extract, adding the extraction solvent to the test product extract, filtering, and taking the filtrate as the test product solution;

[0091] The test solution is injected into an ultra-high performance liquid chromatograph, the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, and a chromatogram of the test product is obtained, and the chromatogram of the test product is compared with the characteristic spectrum of white lentils obtained by the above-mentioned construction method;

[0092] The chromatographic conditions set on the ultra-high performance liquid chromatograph are as described above.

[0093] Optionally, the extraction solvent is an ethanol-water solution. Further optionally, the volume concentration of ethanol in the ethanol-water solution is 60% to 80%. For example, the volume concentration of ethanol in the ethanol-water solution is 60%, 65%, 70%, 75%, or 80%.

[0094] Optionally, the extraction process includes a heating reflux process. Further, optionally, the heating reflux process lasts for 20 to 40 minutes. For example, the heating reflux process lasts for 20, 25, 30, 35, or 40 minutes. Further, optionally, the heating reflux process is performed at a temperature of 80 to 90°C. For example, the heating reflux process is performed at a temperature of 80°C, 85°C, or 90°C.

[0095] Optionally, the extraction process further comprises the following steps:

[0096] After the heating and reflux treatment, the mixture is filtered to obtain a reaction residue and an extract, and the extract is dried to obtain the extract of the test product.

[0097] Optionally, the extraction process further comprises the following steps:

[0098] After the heating and reflux treatment, the mixture is filtered to obtain a reaction residue and an extract, the reaction residue is washed with the extraction solvent to obtain a washing solution, the extract and the washing solution are mixed, and dried to obtain the extract of the test product.

[0099] Optionally, the counterfeit white lentil bean is at least one of white kidney bean, lentil, white eyebrow bean and white sword bean.

[0100] The characteristic spectrum of the processed white lentil product constructed above can be used to assess the quality of the processing method. That is, the quality of the processed white lentil product can be judged based on the characteristic spectrum of the processed white lentil product to reflect the quality of the processing method. A third aspect of the present invention provides a method for detecting a processed white lentil product, comprising the following steps:

[0101] Extracting the processed product to be tested with an extraction solvent to obtain an extract of the processed product to be tested, adding the extraction solvent to the extract of the processed product to be tested, filtering, and taking the filtrate as a solution of the processed product to be tested;

[0102] The solution of the processed product to be tested is injected into an ultra-high performance liquid chromatograph, the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, and a chromatogram of the processed product to be tested is obtained, and the chromatogram of the processed product to be tested is compared with the characteristic spectrum of the processed white lentil obtained by the above-mentioned construction method;

[0103] The chromatographic conditions set on the ultra-high performance liquid chromatograph are as described above.

[0104] Optionally, the extraction solvent is an ethanol-water solution. Further optionally, the volume concentration of ethanol in the ethanol-water solution is 60% to 80%. For example, the volume concentration of ethanol in the ethanol-water solution is 60%, 65%, 70%, 75%, or 80%.

[0105] Optionally, the extraction process includes a heating reflux process. Further, optionally, the heating reflux process lasts for 20 to 40 minutes. For example, the heating reflux process lasts for 20, 25, 30, 35, or 40 minutes. Further, optionally, the heating reflux process is performed at a temperature of 80 to 90°C. For example, the heating reflux process is performed at a temperature of 80°C, 85°C, or 90°C.

[0106] Optionally, the extraction process further comprises the following steps:

[0107] After the heating and reflux treatment, the mixture is filtered to obtain a reaction residue and an extract, and the extract is dried to obtain the extract of the processed product to be tested.

[0108] Optionally, the extraction process further comprises the following steps:

[0109] After the heating and reflux treatment, the reaction residue and the extract are filtered to obtain a reaction residue and an extract, the reaction residue is washed with the extraction solvent to obtain a washing solution, the extract and the washing solution are mixed, and dried to obtain the extract of the prepared product to be tested.

[0110] Optionally, the processed white lentil product is fried white lentil.

[0111] The embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. For the experimental methods in the following examples where specific conditions are not specified, reference should be made to the instructions given in the present invention, or to the experimental manuals or conventional conditions in the field, or to the conditions recommended by the manufacturer, or to experimental methods known in the art. In the following specific examples, the measurement parameters of the raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operating accuracy are allowed. In the following specific examples, 70% ethanol and 75% ethanol refer to ethanol aqueous solution with a volume concentration of 70% and ethanol aqueous solution with a volume concentration of 75%, respectively, 60% methanol refers to methanol aqueous solution with a volume concentration of 60%, 0.4% acetic acid solution refers to acetic acid aqueous solution with a volume concentration of 0.4%,

[0112] Example 1

[0113] This embodiment provides a method for constructing a characteristic spectrum of white dwarf bean medicinal material or decoction pieces, and the specific process is as follows:

[0114] 1. Instruments, reagents and test drugs

[0115] The instruments used were an Agilent ultra-high performance liquid chromatograph (Agilent 1290), an evaporative light scattering detector (Alltech 6000, Aotai Technology Co., Ltd.), a Waters HSS T3 (3 mm × 150 mm, 1.8 μm) chromatographic column, a ten-thousandth balance (ME204E, Mettler-Toledo), a millionth balance (XP26, Mettler-Toledo), an electronic balance (JJ600, Changshu Shuangjie Testing Instrument Factory), a digitally controlled ultrasonic cleaner (KQ-500DE, Kunshan Ultrasonic Instrument Co., Ltd.), a constant temperature water bath (HWS28, Shanghai Yiheng Technology Co., Ltd.), and an ultrapure water system (Milli-Q Direct, Merck).

[0116] Reagents: ethanol (Xilong Scientific Co., Ltd.) and methanol (Xilong Scientific Co., Ltd.) were of analytical grade; acetonitrile (Merck Co., Ltd.) and acetic acid (Tianjin Komiou Chemical Reagent Co., Ltd.) were of HPLC grade; and water was ultrapure water (prepared in the laboratory).

[0117] Test drugs: Panax notoginseng saponin IVa (batch number: 111861-201102, content 77.6%, China Food and Drug Inspection Institute); Copteroside G (batch number: FH22A061, content: 98.0%, Chengdu Pusi Biotechnology Co., Ltd.); The batch number information of 14 batches of white hyacinth bean is shown in Table 1 (since the white hyacinth bean medicinal material and the white hyacinth bean decoction pieces do not need to be processed, the medicinal materials are directly used for decoction pieces, so there is no need to distinguish them).

[0118] Table 1

[0119]

[0120] 2. Preparation of test solution

[0121] Take about 6.0 g of the powder of this product (passed through No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 50 mL of 70% ethanol, weigh it, heat it to 85°C, reflux it for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 70% ethanol, filter it to get the reaction residue and extract, wash the container and the reaction residue with 10 mL of 70% ethanol in batches, combine the washing liquid with the extract, evaporate to dryness, get the extract, add 70% ethanol to dissolve it and transfer it to a 5 mL volumetric flask, add 70% ethanol to the scale, shake it well, centrifuge it (at 12,000 revolutions per minute) for 5 minutes, take the supernatant, filter it, and take the filtrate to get it.

[0122] 3. Preparation of Reference Solution

[0123] Take appropriate amounts of Panax notoginseng saponin IVa reference substance and Copteroside G reference substance, accurately weigh them, and add methanol to prepare reference substance solutions containing 0.2 mg of Panax notoginseng saponin IVa per 1 mL and 0.1 mg of Copteroside G per 1 mL, respectively.

[0124] 4. Chromatographic conditions

[0125] A Waters HSS T3 column (150 mm length, 3 mm inner diameter, 1.8 μm particle size) was used; mobile phase A was acetonitrile, and mobile phase B was 0.4% acetic acid solution. Gradient elution was performed according to the elution program in Table 2. The flow rate was 0.60 mL / min, and the column temperature was 40°C. Detection was performed using an evaporative light scattering detector (carrier gas flow rate: 2.9 L / min; drift tube temperature: 106°C). The number of theoretical plates, calculated based on the ginsenoside IVa peak, should be no less than 4000.

[0126] Table 2

[0127]

[0128] 5. Construction of characteristic maps of white lentil medicinal materials or decoction pieces

[0129] Referring to the method under item "2", 14 batches of white lentil test solution were prepared. Accurately aspirate 5 μl of each of the 14 batches of white lentil test solution and the reference solution under item "3", inject them into the ultra performance liquid chromatograph, and determine according to the chromatographic conditions under item "4" to obtain the chromatograms of the 14 batches of white lentil and the reference solution.

[0130] The “Chinese Herbal Medicine Chromatographic Characteristic Spectrum Similarity Evaluation System (2012.0 Version)” was used to process the chromatogram data of 14 batches of white lentils to generate superimposed spectra. Figure 1 , 14 common peaks with relatively stable relative retention time and relative peak area RSD were selected as characteristic peaks, and the control characteristic spectrum was generated. Figure 2 Copteroside G and Panax notoginseng saponin IVa reference material identification diagram Figure 3 .

[0131] 6. Determination of the characteristic spectrum of white lentil medicinal materials or decoction pieces

[0132] The characteristic spectra of 14 batches of white lentil were analyzed. The chromatographic peak 6 corresponding to Copteroside G was used as the reference peak S1, and the relative retention times and relative peak areas of peaks 1 to 10 were calculated. The chromatographic peak 11 corresponding to ginsenoside IVa was used as the reference peak S2, and the relative retention times and relative peak areas of peaks 12 to 14 were calculated, and the RSD values ​​were calculated. The results are shown in Tables 3 and 4. Table 3 shows the relative retention times of the characteristic peaks in the characteristic spectra of 14 batches of white lentil, and Table 4 shows the relative peak areas of the characteristic peaks in the characteristic spectra of 14 batches of white lentil.

[0133] Table 3

[0134]

[0135] Table 4

[0136]

[0137]

[0138] The results showed that the characteristic spectra of 14 batches of white lentil medicinal materials / decoction pieces had 14 common peaks, with chromatographic peak 6 corresponding to Copteroside G as reference peak S1, and chromatographic peak 11 corresponding to ginsenoside IVa as reference peak S2. The relative retention time RSD values ​​of the other 12 characteristic peaks of the characteristic spectra of 14 batches of white lentil medicinal materials / decoction pieces were between 0.07% and 0.50%, which met the method requirements of the characteristic spectra of white lentil medicinal materials / decoction pieces; the relative peak area RSD values ​​of the remaining 12 characteristic peaks of the characteristic spectra of 14 batches of white lentil were between 20.49% and 75.72%. The results showed that there were certain differences in the corresponding components of the characteristic peaks of white lentils from different origins.

[0139] Therefore, the characteristic spectrum standard of white lentil medicinal materials / decoction pieces is stipulated as follows: 14 characteristic peaks should be present in the chromatogram of the test sample, of which 2 peaks should correspond to the retention time of the reference peaks of the corresponding reference substances; the peak corresponding to the Copteroside G reference substance is the S1 peak, and the relative retention times of peaks 1 to 10 and the S1 peak are calculated; the peak corresponding to the ginsenoside Iva reference substance is the S2 peak, and the relative retention times of peaks 12 to 14 and the S2 peak are calculated, and the relative retention times should be within the range of ±10% of the specified values, and the specified values ​​are: 0.59 (peak 1), 0.66 (peak 2), 0.78 (peak 3), 0.86 (peak 4), 0.91 (peak 5), 1.08 (peak 7), 1.11 (peak 8), 1.20 (peak 9), 1.24 (peak 10), 1.02 (peak 12), 1.05 (peak 13), and 1.09 (peak 14).

[0140] Example 2

[0141] This embodiment provides a method for constructing a characteristic spectrum of stir-fried white lentil slices, and the specific process is as follows:

[0142] 1. Instruments, reagents and test drugs

[0143] According to the 2020 edition of the Chinese Pharmacopoeia, stir-fry white lentils (14 batches) in Table 1 according to the stir-fry method (General Rule 0213) until slightly yellow with burnt spots. Process parameters: Remove impurities from the white lentils and stir-fry at 120°C for 10-15 minutes to obtain 14 batches of stir-fried white lentil slices.

[0144] For other information, please refer to the instruments, reagents and drugs under item "1" in Example 1.

[0145] 2. Preparation of the test solution Refer to the preparation of the test solution under "2" in Example 1.

[0146] 3. Preparation of reference solution Refer to the preparation of reference solution under "3" in Example 1.

[0147] 4. Chromatographic conditions Refer to the chromatographic conditions under item "4" in Example 1.

[0148] 5. Construction of characteristic spectrum of stir-fried white lentil slices

[0149] Referring to the method under item "2", 14 batches of fried white lentil test solutions were prepared. 5 μl of each of the 14 batches of fried white lentil test solutions and the reference solution under item "3" were accurately aspirated and injected into the ultra performance liquid chromatograph. Referring to the chromatographic conditions under item "4", the chromatograms of the 14 batches of fried white lentil and the reference solution were obtained.

[0150] The chromatogram data of 14 batches of stir-fried white lentils were processed using the “Chinese Herbal Medicine Chromatographic Characteristic Spectrum Similarity Evaluation System (2012.0 Version)” to generate superimposed spectra. Figure 4 , 13 common peaks with relatively stable relative retention time and relative peak area RSD were selected as characteristic peaks, and the control characteristic spectrum was generated. Figure 5 .

[0151] 6. Determination of the characteristic spectrum of stir-fried white lentil slices

[0152] The characteristic spectra of 14 batches of fried white lentils were analyzed. The chromatographic peak 6 corresponding to Copteroside G was used as the reference peak S1, and the relative retention times and relative peak areas of peaks 1 to 10 were calculated. The chromatographic peak 11 corresponding to ginsenoside IVa was used as the reference peak S2, and the relative retention times and relative peak areas of peaks 12 and 14 were calculated, and the RSD values ​​were calculated. The results are shown in Tables 5 and 6. Table 5 shows the relative retention times of the characteristic peaks in the characteristic spectra of 14 batches of fried white lentils, and Table 6 shows the relative peak areas of the characteristic peaks in the characteristic spectra of 14 batches of fried white lentils.

[0153] Table 5

[0154]

[0155] Table 6

[0156]

[0157] The results showed that the characteristic spectra of 14 batches of fried white lentil slices had 13 common peaks, with chromatographic peak 6 corresponding to Copteroside G as reference peak S1, and chromatographic peak 11 corresponding to ginsenoside IVa as reference peak S2. The relative retention time RSD values ​​of the other 11 characteristic peaks of the characteristic spectra of 14 batches of fried white lentil slices were between 0.05% and 0.42%, which met the method requirements of the characteristic spectra of fried white lentil slices; the relative peak area RSD values ​​of the other 11 characteristic peaks of the characteristic spectra of 14 batches of fried white lentil slices were between 22.49% and 60.32%. The results showed that there were certain differences in the corresponding components of the characteristic peaks of fried white lentil slices from different origins.

[0158] Table 7 shows a comparison of the average peak areas of 14 characteristic peaks from 14 batches of white lentils and those from stir-fried white lentils. The results show that some chromatographic peaks exhibit significant changes before and after stir-frying. Peak 8 exhibited a significant increase in average peak area after stir-frying, while Peaks 12 and 13 exhibited significant decreases. This suggests that stir-frying has a certain impact on the glycoside composition of white lentils. The most significant change was in Peak 13, which completely disappeared after processing according to the processing technique. Therefore, the presence or absence of Peak 13 can be used to determine whether white lentils have been stir-fried, and thus distinguish between raw and processed white lentils.

[0159] Table 7

[0160]

[0161] Therefore, the characteristic spectrum standard of stir-fried white lentil slices is stipulated as follows: 13 characteristic peaks should appear in the chromatogram of the test sample, of which 2 peaks should correspond to the retention time of the reference peaks of the corresponding reference substances; the peak corresponding to the Copteroside G reference substance is the S1 peak, and the relative retention times of peaks 1 to 10 and the S1 peak are calculated; the peak corresponding to the ginsenoside Iva reference substance is the S2 peak, and the relative retention times of peaks 12 and 14 and the S2 peak are calculated, and the relative retention times should be within the range of ±10% of the specified values, and the specified values ​​are: 0.59 (peak 1), 0.66 (peak 2), 0.78 (peak 3), 0.86 (peak 4), 0.91 (peak 5), 1.08 (peak 7), 1.11 (peak 8), 1.20 (peak 9), 1.24 (peak 10), 1.02 (peak 12), and 1.09 (peak 14).

[0162] Example 3

[0163] This embodiment provides a method for distinguishing white lentils from counterfeit white lentils (white kidney beans, lentils, white eyebrow beans, and white sword beans). The specific process is as follows:

[0164] 1. Instruments, reagents and test drugs

[0165] Test drug: Take 1 portion of white lentil (batch number: BBD3) in Table 1.

[0166] Take the counterfeit white lentils, and see Table 8 for the batch number information.

[0167] Table 8

[0168]

[0169] For other information, please refer to the instruments, reagents and drugs under item "1" in Example 1.

[0170] 2. Preparation of test solution of white lentil and its counterfeit products Refer to the preparation of test solution under "2" in Example 1.

[0171] 3. Chromatographic conditions: See the chromatographic conditions under item "4" in Example 1.

[0172] 4. Chromatogram of counterfeit white lentils

[0173] Referring to the method under "2", prepare 1 portion of white lentil test solution and 5 batches of white lentil counterfeit test solution. Accurately pipette 5 μl of each of 1 portion of white lentil test solution and 5 batches of white lentil counterfeit test solution, inject them into the ultra-high performance liquid chromatograph, and determine them according to the chromatographic conditions under "3". The chromatograms of 1 portion of white lentil and 5 batches of white lentil counterfeit are shown in Figure 1. Figure 6 .

[0174] The results showed that the chromatograms of white kidney beans, lentils, white eyebrow beans, and white sword beans differed significantly from the characteristic spectrum of white lentils. Different varieties could be distinguished based on the number of chromatographic peaks. Therefore, the characteristic spectrum of white lentils constructed in Example 1 can effectively characterize its unique chemical components and effectively distinguish it from its counterfeits, showing a certain degree of specificity.

[0175] Therefore, the identification method of white lentils and their counterfeits is stipulated as follows:

[0176] Take about 6.0 g of the powder to be tested (passed through No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 50 mL of 70% ethanol, weigh it, heat it to 85°C, reflux it for 30 minutes, take it out, let it cool, weigh it again, make up the lost weight with 70% ethanol, filter it, and get the reaction residue and extract. Wash the container and the reaction residue with 10 mL of 70% ethanol in batches, combine the washing liquid with the extract, evaporate it to dryness, get the extract, add 70% ethanol to dissolve it and transfer it to a 5 mL volumetric flask, add 70% ethanol to the scale, shake it well, centrifuge it (at 12,000 revolutions per minute) for 5 minutes, take the supernatant, filter it, and take the filtrate to get the solution of the product to be tested.

[0177] Accurately pipette 5 μl of the test solution and inject it into an ultra-high performance liquid chromatograph. Refer to the chromatographic conditions under item "3" to obtain a chromatogram of the test product. Compare the chromatogram of the test product with the characteristic spectrum of white lentils obtained by the above-mentioned construction method. Determine whether the test powder is white lentils or its counterfeit based on the number of chromatographic peaks.

[0178] Example 4

[0179] This embodiment provides a method for detecting fried white lentils. According to the characteristic spectrum of the fried white lentils, the quality of the fried white lentils is judged to reflect the quality of the frying method. The specific process is as follows:

[0180] Stir-fried white lentil slices were prepared according to the processing regulations for white lentil in Part I of the 2020 edition of the Chinese Pharmacopoeia. The specific processing method is as follows: clean white lentils are stir-fried according to the stir-frying method (General Rules 0213, Part IV, 2020 edition of the Chinese Pharmacopoeia) until they are slightly yellow with burnt spots. This example examines the correspondence between the surface characteristics of stir-fried white lentils and the characteristic spectrum of stir-fried white lentils when the temperature is 80°C to 140°C and the stir-frying time is 5 minutes to 25 minutes.

[0181] 1. Instruments, reagents and test drugs

[0182] Test drug: Take 1 portion of white lentil (batch number: BBD3) in Table 1.

[0183] Take 4 portions of white lentils (batch number: BBD3) in Table 1, weigh about 50 g each, remove impurities, and place them in a preheated wok. According to the stir-fry method (General Chapter 0213 of Part 4 of the Chinese Pharmacopoeia 2020 Edition), heat them to 80°C, 100°C, 120°C, and 140°C for 10 minutes to obtain 4 portions of stir-fried white lentils at different stir-frying temperatures.

[0184] Take 5 portions of white lentils (batch number: BBD3) in Table 1, weigh about 50 g each, remove impurities, and place them in a preheated wok. According to the stir-fry method (General Chapter 0213 of Part 4 of the Chinese Pharmacopoeia 2020 Edition), heat to 120°C over low heat and stir-fry for 5 minutes, 10 minutes, 15 minutes, 20 minutes, and 25 minutes to obtain 5 portions of stir-fried white lentils at different stir-frying times.

[0185] For other information, please refer to the instruments, reagents and drugs under item "1" in Example 1.

[0186] 2. Preparation of the test solution Refer to the preparation of the test solution under "2" in Example 1.

[0187] 3. Chromatographic conditions: See the chromatographic conditions under item "4" in Example 1.

[0188] 4. Correspondence between the surface characteristics of fried white lentils and the characteristic spectrum of fried white lentils

[0189] 4.1 Corresponding situations of frying at different temperatures

[0190] The initial surface characteristics of white lentils and the surface characteristics of fried white lentils after frying at different temperatures for 10 minutes were recorded. The results are shown in Tables 9 and Figure 7 .

[0191] Table 9

[0192]

[0193] Refer to the method under "2" to prepare 1 portion of white lentil test solution and the above 4 portions of fried white lentil test solutions at different frying temperatures. Accurately pipette 5 μl of each of 1 portion of white lentil test solution and 4 portions of fried white lentil test solutions, inject them into the ultra-high performance liquid chromatograph, and determine the chromatographic conditions under "3" to obtain 1 portion of white lentil chromatogram and 4 portions of fried white lentil chromatogram, see Figure 8 Among them, the peak areas of each chromatographic peak in the chromatograms of 1 portion of white lentils and 4 portions of fried white lentils are shown in Table 10.

[0194] Table 10

[0195]

[0196] From Table 9 and Figure 7 It can be seen that when stir-fried at 80°C and 100°C for 10 minutes, the white lentils had no burnt spots on the surface. When stir-fried at 120°C for 10 minutes, the color of the stir-fried white lentils was more consistent with the pharmacopoeial properties: a slightly yellow surface with a bean aroma, burnt spots, and a slightly yellow cross-section. When the stir-frying temperature was increased to 140°C, the color of the stir-fried white lentils gradually deepened, with some parts turning dark yellow and having a burnt aroma, approaching a burnt state, which did not meet the pharmacopoeial description of "slightly yellow surface with burnt spots." Therefore, the stir-frying temperature should not be too high, and 120°C is appropriate.

[0197] From Table 10 and Figure 8 It can be seen that with the increase of frying temperature, the peak areas of different chromatographic peaks increase or decrease to varying degrees, especially after frying at 120°C and 140°C for 10 minutes, Peak 13 completely disappears, which is consistent with the results in Example 2. It also explains that the reason for the disappearance of Peak 13 may be due to thermal destruction or transformation during the frying process.

[0198] 4.2 Corresponding situations of stir-frying at different times

[0199] The initial surface characteristics of white lentils and the surface characteristics of white lentils after frying at 120℃ for different times were recorded. The results are shown in Tables 11 and Figure 9 .

[0200] Table 11

[0201]

[0202] Referring to the method under "2", prepare 1 portion of white lentil test solution and the above 5 portions of fried white lentil test solutions at different frying times. Accurately pipette 5 μl of each of 1 portion of white lentil test solution and 5 portions of fried white lentil test solutions, inject them into the ultra-high performance liquid chromatograph, and determine the chromatographic conditions under "3" to obtain the chromatograms of 1 portion of white lentil and 5 portions of fried white lentil. Figure 10Among them, the peak areas of each chromatographic peak in the chromatograms of 1 portion of white lentils and 5 portions of fried white lentils are shown in Table 12.

[0203] Table 12

[0204]

[0205] From Table 11 and Figure 9 As can be seen, at a frying temperature of 120°C and at different frying times, the surface color of the stir-fried white lentil slices gradually deepens, a fragrant aroma emerges, and the number of burnt spots increases with increasing heating time. The 10-15 minute frying time best matches the pharmacopoeial description of stir-fried white lentils. Combined with the Chinese Pharmacopoeia's requirement for stir-fried white lentils to be "slightly yellow with burnt spots," the specific preparation method for stir-fried white lentil slices is as follows: fry the white lentil slices at 120°C for 10-15 minutes.

[0206] From Table 12 and Figure 10 It can be seen that when the frying temperature is 120°C, peak 13 completely disappears after 10 minutes as the frying time increases, which is consistent with the results of Example 2. Other chromatographic peaks, such as peak 12, decrease to a certain level after frying and no longer change. After frying for 20 minutes, the peak area of ​​peak 8 does not increase significantly.

[0207] In summary, the characteristic spectrum of stir-fried white lentils is basically consistent with the properties of stir-fried white lentils specified in the Pharmacopoeia. In addition, each chromatographic peak has a trend of change to varying degrees during the stir-frying process, with Peak 13 showing the most obvious change. Based on the premise that the properties specified in the Pharmacopoeia are met as a reference, Peak 13 disappears after processing, which is consistent with the results of Example 2. Therefore, it can be used as a marker for its processing endpoint.

[0208] Therefore, the detection method for fried white lentils is stipulated as follows:

[0209] About 6.0 g of the powder of the processed product to be tested (passed through a No. 3 sieve) was accurately weighed and placed in a stoppered conical flask. 50 mL of 70% ethanol was accurately added and weighed. The product was heated to 85°C and refluxed for 30 minutes. The product was removed, cooled, and weighed again. The weight loss was made up with 70% ethanol. The product was filtered to obtain a reaction residue and an extract. The container and the reaction residue were washed with 10 mL of 70% ethanol in portions. The washing solution was added to the extract and evaporated to dryness to obtain an extract. The extract was dissolved in 70% ethanol and transferred to a 5 mL volumetric flask. 70% ethanol was added to the mark, shaken, and centrifuged (at 12,000 rpm) for 5 minutes. The supernatant was collected and filtered. The filtrate was then collected to obtain the solution of the processed product to be tested.

[0210] Accurately pipette 5 μl of the solution of the prepared product to be tested and inject it into the ultra-high performance liquid chromatograph. Refer to the chromatographic conditions under item "3" to obtain the chromatogram of the prepared product to be tested. Compare the chromatogram of the prepared product to be tested with the characteristic spectrum of the white lentil prepared product obtained by the above-mentioned construction method. Judge whether the prepared product to be tested is of qualified quality based on the presence or absence of peak 13 and whether it meets the properties specified in the pharmacopoeia.

[0211] Comparative Example 1

[0212] This comparative example provides a method for constructing a characteristic spectrum of white lentils, which is as follows:

[0213] 1. Instruments, reagents and test drugs

[0214] Instrument: Waters ultra-high performance liquid chromatograph (H-class, Waters Corporation), UV detector (PDA).

[0215] Test drug: Take 1 portion of white lentil (batch number: BBD1) in Table 1.

[0216] For other information, please refer to the instruments, reagents and drugs under item "1" in Example 1.

[0217] 2. Preparation of the test solution Refer to the preparation of the test solution under "2" in Example 1.

[0218] 3. Chromatographic conditions

[0219] The detection wavelength is 210 nm.

[0220] For other conditions, see chromatographic conditions under item "4" in Example 1

[0221] 4. Determination

[0222] Prepare 1 portion of white lentil test solution by referring to the method under "2". Accurately pipette 5 μl of 1 portion of white lentil test solution and inject it into the ultra-high performance liquid chromatograph. Determine according to the chromatographic conditions under "3". Obtain 1 portion of white lentil chromatogram, see Figure 11 .

[0223] contrast Figure 11 and Figure 2 (The white lentil control characteristic spectrum of Example 1) shows that the number of chromatographic peaks in the chromatogram obtained using the UV detector is relatively small. This may be because some components have a low or no response under the UV detector conditions, which cannot well reflect the overall quality of the white lentil. The evaporative light detector used in Example 1 has a better response to glycosides and saponins, which is conducive to the construction of the white lentil characteristic spectrum.

[0224] Comparative Example 2

[0225] This comparative example provides a method for constructing a characteristic spectrum of white lentils, which is as follows:

[0226] 1. Instruments, reagents and test drugs

[0227] Test drug: Take 1 portion of white lentil (batch number: BBD12) in Table 1.

[0228] For other information, please refer to the instruments, reagents and drugs under item "1" in Example 1.

[0229] 2. Preparation of test solution

[0230] Take about 6.0 g of the powder of this product (BBD12, passed through No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, add 50 mL of 75% ethanol, ultrasonically treat it (power 300 W, frequency 40 kHz) for 30 minutes, let it cool, filter it to obtain the reaction residue and extract, wash the container and the reaction residue with a small amount of 75% ethanol in batches, combine the washing liquid with the extract and concentrate it to dryness to obtain the extract, add 3 mL of water to dissolve it, pass it through a hydrophilic-lipophilic balance solid phase extraction column (1 g, 6 mL, pre-washed with 12 mL of methanol first, then pre-washed with 12 mL of water), elute it with 10 mL of 60% methanol, discard the eluate, and then elute it with 9 mL of methanol into a 10 mL volumetric flask, add methanol to the scale, shake well, and obtain it.

[0231] 3. Chromatographic conditions refer to the chromatographic conditions under item "4" in Example 1

[0232] 4. Determination

[0233] Prepare 1 portion of white lentil test solution by referring to the method under "2". Accurately pipette 5 μl of 1 portion of white lentil test solution and inject it into the ultra-high performance liquid chromatograph. Determine according to the chromatographic conditions under "3". Obtain 1 portion of white lentil chromatogram, see Figure 12 .

[0234] contrast Figure 12 and Figure 2 (Comparative characteristic spectrum of white lentil in Example 1) It can be seen that 14 chromatographic peaks were barely detected in this comparative example, but except for peak 11, the peak areas of other chromatographic peaks were relatively small, and peak 13 was almost invisible, which could not achieve the purpose of distinguishing white lentil from its processed products.

[0235] Comparative Example 3

[0236] This comparative example provides a method for constructing a characteristic spectrum of white lentils, which is as follows:

[0237] 1. Instruments, reagents and test drugs

[0238] Test drug: Take 1 portion of white lentil (batch number: BBD1) in Table 1.

[0239] For other information, please refer to the instruments, reagents and drugs under item "1" in Example 1.

[0240] 2. Preparation of test solution: See the preparation of test solution under "2" in Example 1.

[0241] 3. Chromatographic conditions

[0242] Acetonitrile was used as mobile phase A, and 10 mmol ammonium acetate solution was used as mobile phase B.

[0243] For other conditions, see chromatographic conditions under item "4" in Example 1

[0244] 4. Determination

[0245] Prepare 1 portion of white lentil test solution by referring to the method under "2". Accurately pipette 5 μl of 1 portion of white lentil test solution and inject it into the ultra-high performance liquid chromatograph. Determine according to the chromatographic conditions under "3". Obtain 1 portion of white lentil chromatogram, see Figure 13 .

[0246] contrast Figure 13 and Figure 2 (The comparative characteristic spectrum of white lentil in Example 1) It can be seen that the chromatogram of this comparative example basically only shows one main peak of ginsenoside IVa, and the responses of other chromatographic peaks are very low under this condition.

[0247] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0248] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A method for constructing a characteristic spectrum of white lentil or its processed product, characterized in that: The following steps are involved: extracting white lentil or its processed product with an extraction solvent to obtain an extract, adding the extraction solvent to the extract, filtering, and taking the filtrate as a test solution, wherein the white lentil is a white lentil medicinal material or a white lentil decoction piece, the processed white lentil is fried white lentil, and the extraction solvent is an ethanol aqueous solution; The test sample solution is injected into an ultra-high performance liquid chromatograph, the detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, and a chromatogram of the test sample is obtained. The chromatogram of the test sample is imported into a traditional Chinese medicine chromatographic fingerprint similarity evaluation software to obtain a characteristic spectrum of white lentil or its processed product, wherein the characteristic spectrum of white lentil has a total of 14 characteristic peaks, including characteristic peaks of Copteroside G and Panax notoginseng saponin Iva; the characteristic spectrum of the processed white lentil has a total of 13 characteristic peaks, including characteristic peaks of Copteroside G and Panax notoginseng saponin Iva; The chromatographic conditions set by the ultra-high performance liquid chromatograph include: The mobile phase includes mobile phase A and mobile phase B, wherein mobile phase A includes acetonitrile and mobile phase B includes an acetic acid aqueous solution, wherein the volume concentration of acetic acid in the acetic acid aqueous solution is 0.2% to 0.6%. The chromatographic column is a Waters HSS T3 column with a column length of 150 mm, an inner diameter of 3 mm, and a particle size of 1.8 μm. The elution procedure includes: From 0 min to 17 min, the volume percentage of mobile phase A increased from 25% to 29%, and the volume percentage of mobile phase B decreased from 75% to 71%; From 17 min to 25 min, the volume percentage of mobile phase A was maintained at 29%, and the volume percentage of mobile phase B was maintained at 71%; From 25 to 43 minutes, the volume percentage of mobile phase A increased from 29% to 41%, and the volume percentage of mobile phase B decreased from 71% to 59%; From 43 to 46 minutes, the volume percentage of mobile phase A increased from 41% to 90%, and the volume percentage of mobile phase B decreased from 59% to 10%; From 46 min to 49 min, the volume percentage of mobile phase A was maintained at 90%, and the volume percentage of mobile phase B was maintained at 10%.

2. The construction method according to claim 1, characterized in that The flow rate is 0.40 mL to 0.80 mL per minute.

3. The construction method according to claim 1, characterized in that The column temperature is 38℃~42℃.

4. The construction method according to claim 1, characterized in that The carrier gas flow rate of the evaporative light scattering detector is 2.7 L / min to 3.1 L / min.

5. The construction method according to claim 1, characterized in that The drift tube temperature of the evaporative light scattering detector is 102° C. to 110° C.

6. The construction method according to any one of claims 1 to 5, characterized in that: The extraction process includes a heating reflux process.

7. A method for detecting a processed white lentil product, characterized in that: The following steps are involved: Extracting the processed product to be tested with an extraction solvent to obtain an extract of the processed product to be tested, adding the extraction solvent to the extract of the processed product to be tested, filtering, and taking the filtrate as a solution of the processed product to be tested, wherein the processed product to be tested is stir-fried white lentils to be tested, and the extraction solvent is an ethanol aqueous solution; Injecting the solution of the processed product to be tested into an ultra-high performance liquid chromatograph using an evaporative light scattering detector as the detector of the ultra-high performance liquid chromatograph to obtain a chromatogram of the processed product to be tested, and comparing the chromatogram of the processed product to be tested with the characteristic spectrum of the processed white lentil obtained by the construction method according to any one of claims 1 to 6; The chromatographic conditions set in the ultra-high performance liquid chromatograph are as described in any one of claims 1 to 5.

8. The detection method according to claim 7, characterized in that The extraction process includes a heating reflux process.

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