Construction method and application of specific chromatogram of white hyacinth bean or processed product thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine analysis, in particular to a construction method and application of a specific chromatogram of white hyacinth beans or processed products thereof. The construction method of the characteristic spectrum of the white hyacinth bean or the processed product thereof comprises the following steps: extracting the white hyacinth bean or the processed product thereof by using an extraction solvent to obtain an extract, adding the extraction solvent into the extract, filtering, and taking a subsequent filtrate as a test solution; the sample solution is injected into an ultra-high performance liquid chromatograph, a detector of the ultra-high performance liquid chromatograph is an evaporative light scattering detector, a chromatogram of the sample is obtained, the chromatogram of the sample is imported into traditional Chinese medicine chromatographic fingerprint similarity evaluation software, and the characteristic chromatogram of the white hyacinth bean or the processed product of the white hyacinth bean is obtained. The specific chromatogram of the white hyacinth bean has 14 characteristic peaks, the specific chromatogram of the processed product of the white hyacinth bean has 13 characteristic peaks, and the white hyacinth bean and the processed product of the white hyacinth bean can be distinguished according to the specific chromatogram of the white hyacinth bean or the specific chromatogram of the processed product of the white hyacinth bean.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine analysis, and in particular to a method for constructing a characteristic spectrum of white lentil or its processed product and an application thereof, and in particular to a method for constructing a characteristic spectrum of white lentil or its processed product, a method for identifying white lentil and its counterfeits, and a method for detecting white lentil processed products. Background Art

[0002] White lentil is the dried mature seed of Dolichos ala L., a plant of the legume family. The mature fruits are harvested in autumn and winter, dried in the sun, the seeds are removed, and then dried again. White lentil tastes sweet and is slightly warm in nature; it enters the spleen and stomach meridians. It has the effects of strengthening the spleen and eliminating dampness, harmonizing the middle and relieving heat. It is used to treat spleen and stomach weakness, loss of appetite, loose stools, excessive leucorrhea, vomiting and diarrhea due to summer heat and dampness, chest tightness and abdominal distension. White lentil mainly contains nutrients such as protein, fat, polysaccharides, calcium, iron, phosphorus, vitamins, etc., in addition to steroids, glycosides, fenugreek and other ingredients.

[0003] There are differences in the efficacy of white lentils and their processed products. For example, fried white lentils are a processed product of white lentils. In terms of efficacy, the efficacy of white lentils and fried white lentils are different. Raw lentils tend to dispel dampness, clear away heat and nourish the stomach; fried lentils tend to be astringent, invigorate the spleen and stop diarrhea. White lentils have antibacterial and antiviral effects, and enhance immune function. They are used for poor appetite, loose stools, excessive leucorrhea, vomiting and diarrhea due to summer heat, chest tightness and abdominal distension, as well as water retention and thirst. Fried white lentils have a better therapeutic effect on human loss of appetite and weak spleen and stomach. Although fried white lentils can be distinguished based on their properties of "slightly yellow with burnt spots", since white lentils are used as both medicine and food, they are often powdered when used as meal substitutes in food. After being crushed, they are difficult to distinguish based on their properties and are easy to confuse. In addition, some Chinese patent medicines contain white lentil or fried white lentil powder. In view of the certain differences in their medicinal effects, in order to ensure the safety and effectiveness of the medication, it is necessary to establish characteristic spectra of white lentils or characteristic spectra of fried white lentils that can distinguish the two. Summary of the invention

[0004] Based on this, the present invention provides a method for constructing a characteristic spectrum of white hyacinth bean or its processed product and its application.

[0005] The first aspect of the present invention provides a method for constructing a characteristic spectrum of white lentil or its processed product, and the technical scheme thereof is as follows:

[0006] A method for constructing a characteristic spectrum of white lentil or its processed product, comprising the following steps:

[0007] 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;

[0008] 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 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] Wherein, 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 is 0.40 mL to 0.80 mL per minute;

[0020] 3) Column temperature is 38℃~42℃;

[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 ethanol aqueous solution;

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

[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 identifying white lentils and their counterfeits comprises the following steps:

[0029] Extract the sample with an extraction solvent to obtain a sample extract, add the extraction solvent to the sample extract, filter, and take the filtrate as a sample solution;

[0030] The test product 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, a chromatogram of the test product is obtained, and the chromatogram of the test product is compared with the characteristic spectrum of white lentil obtained by the above construction method;

[0031] The chromatographic conditions set in 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 ethanol aqueous solution;

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

[0035] In some embodiments, the white lentil bean fake 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 processed white hyacinth bean products, and the technical scheme thereof is as follows:

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

[0038] Extract the processed product to be tested with an extraction solvent to obtain an extract of the processed product to be tested, add the extraction solvent to the extract of the processed product to be tested, filter, and take 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, 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 in 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 ethanol aqueous solution;

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

[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 method has strong specificity, can more comprehensively reflect the overall quality of white lentils or white lentil processed products, ensure the safety of white lentils or white lentil processed products, and provide 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 more completely understand the present invention and its beneficial effects, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0047] Figure 1 This is an overlay of characteristic spectra of 14 batches of white hyacinth bean 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 lentil and its counterfeit products;

[0053] Figure 7 A real-life picture of fried white lentils cooked at different temperatures;

[0054] Figure 8 It is a comparison diagram of the characteristic spectra of white lentil and fried white lentil at different temperatures;

[0055] Fig. 9 It is a real-time picture of stir-fried white lentils stir-fried at different times;

[0056] Fig.10 It is a comparison diagram of characteristic spectra of white lentil and fried white lentil fried at different times;

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

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

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

[0060] The present invention will be further described in detail below in conjunction with 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 make the disclosure of the present invention more thoroughly understood.

[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 the specification of the present invention herein 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 spectrum of white lentil formula granules is the main one. There are currently 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 spectrum, it cannot distinguish between white lentil and fried white lentil formula granules, and the main parameters of the characteristic spectrum are not provided. The second type is the local standards of other provinces and cities except Shanghai, with a total of 4 to 5 characteristic peaks, identifying fenugreek alkaloids and other amino acids, but 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 lentil mainly focuses on chemical composition, clinical application and pharmacology, and there are few literatures on the quality control research of white lentil. The quality control standards of white lentil in the 2020 edition of the Chinese Pharmacopoeia only include properties, microscopic identification, and moisture inspection, 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), fenugreek alkaloids, and ginsenoside IVa in white lentil, 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 genuine white lentil medicinal materials, it seriously affects the safety of medication. Therefore, it is necessary to establish a characteristic spectrum for the chemical composition of white lentil as a standard for its qualitative identification.

[0064] In order to fill the gap in the characteristic spectrum for distinguishing white lentil and its processed products, the first aspect of the present invention provides a method for constructing a characteristic spectrum of white lentil or its processed products, which comprises 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 a test solution;

[0066] 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 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] Wherein, 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 solid phase extraction column was used to elute the above liquid to obtain the 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, in the quality standard of "White Lentil Formula Granules" issued by Shanghai, an ultraviolet detector is used, 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 specificity is strong, and the overall quality of white lentils or white lentil processed products can be more comprehensively reflected, the drug safety of white lentils or white lentil processed products is ensured, and a reference is provided for formulating the quality standards of 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 column 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: the carrier gas flow rate of the evaporative light scattering detector is 2.7 L / min to 3.1 L / min. For example, the carrier gas flow rate is 2.7 L / min, 2.8 L / min, 2.9 L / min, 3.0 L / min, 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 aqueous solution. Further optionally, the volume concentration of ethanol in the ethanol aqueous solution is 60% to 80%. For example, the volume concentration of ethanol in the ethanol aqueous solution is 60%, 65%, 70%, 75%, or 80%.

[0082] Optionally, the extraction process includes a heating reflux process. Further optionally, the time of the heating reflux process is 20 min to 40 min. For example, the time of the heating reflux process is 20 min, 25 min, 30 min, 35 min, 40 min. Further optionally, the temperature of the heating reflux process is 80 to 90 ° C. For example, the temperature of the heating reflux process is 80 ° C, 85 ° C, 90 ° C.

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

[0084] After the heating and reflux treatment, the reaction residue and the extract are obtained by filtration, 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 decoction pieces.

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

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

[0090] Extract the sample with an extraction solvent to obtain a sample extract, add the extraction solvent to the sample extract, filter, and take the filtrate as a sample solution;

[0091] The test product 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, a chromatogram of the test product is obtained, and the chromatogram of the test product is compared with the characteristic spectrum of white lentil obtained by the above construction method;

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

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

[0094] Optionally, the extraction process includes a heating reflux process. Further optionally, the time of the heating reflux process is 20 min to 40 min. For example, the time of the heating reflux process is 20 min, 25 min, 30 min, 35 min, 40 min. Further optionally, the temperature of the heating reflux process is 80 to 90 ° C. For example, the temperature of the heating reflux process is 80 ° C, 85 ° C, 90 ° C.

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

[0096] After the heating and reflux treatment, the reaction residue and the extract are obtained by filtration, 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 reaction residue and the extract are filtered to obtain a reaction residue, the reaction residue is washed with the extraction solvent to obtain a washing solution, the extraction solution and the washing solution are mixed, and dried to obtain the extract of the test product.

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

[0100] The characteristic spectrum of the processed white lentil product constructed above can be used to examine the quality of the processing method, that is, the quality of the processed white lentil product can be judged according to the characteristic spectrum of the processed white lentil product to reflect the quality of the processing method. The third aspect of the present invention provides a method for detecting a processed white lentil product, which comprises the following steps:

[0101] Extract the processed product to be tested with an extraction solvent to obtain an extract of the processed product to be tested, add the extraction solvent to the extract of the processed product to be tested, filter, and take 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, 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 in the ultra-high performance liquid chromatograph are as described above.

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

[0105] Optionally, the extraction process includes a heating reflux process. Further optionally, the time of the heating reflux process is 20 min to 40 min. For example, the time of the heating reflux process is 20 min, 25 min, 30 min, 35 min, 40 min. Further optionally, the temperature of the heating reflux process is 80 to 90 ° C. For example, the temperature of the heating reflux process is 80 ° C, 85 ° C, 90 ° C.

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

[0107] After the heating and reflux treatment, the reaction residue and the extract are obtained by filtration, 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, 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 processed 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 in conjunction with examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. For experimental methods in the following examples where specific conditions are not specified, reference shall be made to the instructions given in the present invention in priority, and may also be made according to the experimental manual or conventional conditions in the field, or according to the conditions recommended by the manufacturer, or with reference 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 hyacinth bean medicinal materials or decoction pieces, and the specific process is as follows:

[0114] 1. Instruments, reagents and test drugs

[0115] Instruments: Agilent ultra-high performance liquid chromatograph (Agilent 1290, Agilent), evaporative light scattering detector (Alltech 6000, Alltech Technology Co., Ltd.), Waters HSS T3 (3 mm × 150 mm, 1.8 um) chromatographic column; one-ten-thousandth balance (ME204E, Mettler-Toledo); one-millionth balance (XP26, Mettler-Toledo); electronic balance (JJ600, Changshu Shuangjie Testing Instrument Factory); CNC ultrasonic cleaner (KQ-500DE, Kunshan Ultrasonic Instrument Co., Ltd.); constant temperature water bath (HWS28, Shanghai Yiheng Technology Co., Ltd.); ultrapure water system (Milli-Q Direct, Merck & Co., Ltd.).

[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; water was ultrapure water (self-made 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 (because 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, put 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, cool it, weigh it again, make up the lost weight with 70% ethanol, filter it, get the reaction residue and the extract, wash the container and the reaction residue with 10 mL of 70% ethanol in batches, add the washing liquid into 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 (speed is 12000 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] Waters HSS T3 (column length: 150 mm, inner diameter: 3 mm, particle size: 1.8 μm) was used as the chromatographic column; acetonitrile was used as the mobile phase A, 0.4% acetic acid solution was used as the mobile phase B, and gradient elution was performed according to the elution program in Table 2; the flow rate was 0.60 mL per minute; the column temperature was 40°C; and the evaporative light scattering detector (carrier gas flow rate: 2.9 L / min; drift tube temperature: 106°C) was used for detection. The theoretical plate number calculated based on the peak of Panax notoginseng saponin IVa should be no less than 4000.

[0126] Table 2

[0127]

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

[0129] Referring to the method under item "2", 14 batches of white lentil test solution were prepared. 5 μl of each of the 14 batches of white lentil test solution and the reference solution under item "3" were accurately pipetted and injected into the ultra-high performance liquid chromatograph. Referring to the chromatographic conditions under item "4", the chromatograms of the 14 batches of white lentil and the reference solution were obtained.

[0130] The “Chinese Medicine Chromatographic Characteristic Spectrum Similarity Evaluation System (2012.0 Version)” was used to process the chromatographic data of 14 batches of white lentil 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 as shown in Figure 2 Copteroside G and Panax notoginseng saponin IVa reference material identification diagram Figure 3 .

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

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

[0133] Table 3

[0134]

[0135] Table 4

[0136]

[0137]

[0138] The results showed that the characteristic spectra of 14 batches of white hyacinth bean 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 14 batches of white hyacinth bean characteristic spectra were between 0.07% and 0.50%, which met the method requirements of the characteristic spectra of white hyacinth bean medicinal materials / decoction pieces; the relative peak area RSD values ​​of the remaining 12 characteristic peaks of the 14 batches of white hyacinth bean characteristic spectra 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 hyacinth beans from different origins.

[0139] Therefore, the characteristic spectrum standard of white hyacinth bean 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 sample; the peak corresponding to the Copteroside G reference sample is the S1 peak, and the relative retention time of peaks 1 to 10 with the S1 peak is calculated; the peak corresponding to the ginsenoside Iva reference sample is the S2 peak, and the relative retention time of peaks 12 to 14 with the S2 peak is calculated, and the relative retention time should be within ±10% of the specified value, 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 hyacinth bean slices, and the specific process is as follows:

[0142] 1. Instruments, reagents and test drugs

[0143] According to the provisions of the 2020 edition of the Chinese Pharmacopoeia on stir-fried white lentils: Take 14 batches of pure white lentils in Table 1 and stir-fry them according to the stir-fry method (General Rule 0213) until they are slightly yellow with burnt spots. Process parameters: Take pure white lentils, remove impurities, stir-fry at 120℃ for 10 to 15 minutes, and 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 See the preparation of reference solution under item "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 pipetted 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 “Chinese Medicine Chromatographic Characteristic Spectrum Similarity Evaluation System (2012.0 Version)” was used to process the chromatogram data of 14 batches of stir-fried white lentils 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 as shown in 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, wherein 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 remaining 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] The average peak areas of 14 characteristic peaks of 14 batches of white lentils and fried white lentils were compared, as shown in Table 7. The results showed that some chromatographic peaks changed significantly before and after frying. After frying, the average peak area of ​​peak 8 increased significantly, while peaks 12 and 13 decreased significantly. This shows that frying has a certain effect on the glycoside components of white lentils. Among them, the most obvious change is characteristic peak 13. After processing according to the processing technology, peak 13 completely disappears. Therefore, it is possible to judge whether white lentils have been fried based on the presence or absence of peak 13, thereby judging the raw and processed white lentils.

[0159] Table 7

[0160]

[0161] Therefore, the characteristic spectrum standard of stir-fried white hyacinth bean slices is stipulated as follows: 13 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 sample; the peak corresponding to the Copteroside G reference sample is the S1 peak, and the relative retention time of peaks 1 to 10 with the S1 peak is calculated; the peak corresponding to the ginsenoside Iva reference sample is the S2 peak, and the relative retention time of peaks 12 and 14 with the S2 peak is calculated, and the relative retention time should be within ±10% of the specified value, 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 identifying white lentils and white lentil counterfeits (white kidney beans, lentils, white eyebrow beans and white sword beans), and the specific process is as follows:

[0164] 1. Instruments, reagents and test drugs

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

[0166] Take the counterfeit white lentil, and its batch number information is shown in Table 8.

[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 hyacinth bean and its counterfeit products Refer to the preparation of test solution under item "2" in Example 1.

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

[0172] 4. Chromatogram of fake white lentil

[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 into ultra-high performance liquid chromatography, and determine 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 show that the chromatograms of white kidney beans, lentils, white eyebrow beans and white sword beans are quite different from the characteristic spectrum of white lentils. Different varieties can be distinguished according to the number of chromatographic peaks. Therefore, the characteristic spectrum of white lentils constructed in Example 1 can effectively characterize its unique chemical components, effectively distinguish it from its counterfeits, and has certain 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, put 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, cool it, weigh it again, make up the lost weight with 70% ethanol, filter it, get the reaction residue and the extract, wash the container and the reaction residue with 10 mL of 70% ethanol in batches, add the washing liquid into 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 (speed is 12000 revolutions per minute) for 5 minutes, take the supernatant, filter it, take the filtrate to get the solution of the product to be tested.

[0177] Accurately pipette 5 μl of the test solution, inject it into the ultra-high performance liquid chromatograph, and determine it according 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 lentil obtained by the above-mentioned construction method, and judge whether the test powder is white lentil or its counterfeit according to 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] The stir-fried white lentil slices were prepared according to the processing regulations of the white lentil item in 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 of Part 4 of the 2020 edition of the Chinese Pharmacopoeia) until they are slightly yellow with burnt spots. This example investigates the correspondence between the surface properties of stir-fried white lentils and the characteristic map of stir-fried white lentils when the slow fire heating 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 hyacinth bean (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 preheated woks respectively. According to the stir-fry method (General Rules 0213 of Part 4 of the Chinese Pharmacopoeia 2020 Edition), heat to 80°C, 100°C, 120°C, and 140°C for 10 minutes over low heat 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 preheated woks respectively. According to the stir-fry method (General Rules 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 Refer to 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] Referring to the method under "2", prepare 1 portion of white lentil test solution and the above 4 portions of stir-fried white lentil test solutions at different stir-frying temperatures. Accurately pipette 5 μl of each of 1 portion of white lentil test solution and 4 portions of stir-fried white lentil test solutions, inject them into the ultra-high performance liquid chromatograph, and determine them according to the chromatographic conditions under "3" to obtain 1 portion of white lentil chromatogram and 4 portions of stir-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 the white lentils are fried at 80℃ and 100℃ for 10 minutes, there are no burnt spots on the surface. When fried at 120℃ for 10 minutes, the color of the fried white lentils is more in line with the pharmacopoeia properties, with a slightly yellow surface, bean aroma, burnt spots, and slightly yellow cross-section. When the frying temperature is increased to 140℃, the color of the fried white lentils gradually deepens, partly dark yellow, with a burnt aroma, and nearly burnt, which does not meet the description of the pharmacopoeia property of "slightly yellow surface with burnt spots". Therefore, the frying temperature should not be too high, and 120℃ 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 heat destruction or transformation during the frying process.

[0198] 4.2 Corresponding situation of frying at different times

[0199] The initial surface characteristics of white lentils and the surface characteristics of white lentils fried at 120°C for different times were recorded. The results are shown in Tables 11 and Fig. 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 stir-fried white lentil test solutions at different stir-frying times. Accurately pipette 5 μl of each of 1 portion of white lentil test solution and 5 portions of stir-fried white lentil test solutions, inject them into the ultra-high performance liquid chromatograph, and determine them according to the chromatographic conditions under "3" to obtain 1 portion of white lentil chromatogram and 5 portions of stir-fried white lentil chromatogram, see Fig.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 Fig. 9 It can be seen that when the frying temperature is 120℃ and the frying time is different, the color of the surface of the fried white lentil slices gradually deepens, the aroma overflows, and the burnt spots increase with the continuous extension of the heating time. Among them, the 10-15 minutes, the properties of fried white lentils are most consistent with the description specified in the pharmacopoeia. Combined with the "Chinese Pharmacopoeia" requirements for the properties of fried white lentils "slightly yellow surface with burnt spots", the specific preparation method of fried white lentil slices is finally obtained: take white lentil slices and fry them at 120℃ for 10-15 minutes.

[0206] From Table 12 and Fig.10 It can be seen that when the frying temperature is 120°C, as the frying time increases, peak 13 disappears completely after 10 minutes, which is consistent with the results of Example 2. Other chromatographic peaks, such as peak 12, decrease to a certain extent 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 fried white lentils is basically consistent with the properties of fried white lentils specified in the pharmacopoeia. In addition, each chromatographic peak has a trend of change to varying degrees during the frying process, and the most obvious change is peak 13. Under the premise of meeting the properties specified in the pharmacopoeia as a reference, peak 13 disappears after processing, which is consistent with the results of Example 2, and can therefore be used as a marker for its processing end point.

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

[0209] Take about 6.0 g of the powder of the processed product to be tested (passed through a 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, get the reaction residue and the extract, wash the container and the reaction residue with 10 mL of 70% ethanol in batches, add the washing liquid into 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 (speed is 12000 revolutions per minute) for 5 minutes, take the supernatant, filter it, take the filtrate, and get the solution of the processed product to be tested;

[0210] Accurately pipette 5 μl of the solution of the processed product to be tested, inject it into the ultra-high performance liquid chromatograph, and determine it according to the chromatographic conditions under item "3" to obtain the chromatogram of the processed product to be tested. Compare the chromatogram of the processed product to be tested with the characteristic spectrum of the processed white hyacinth bean obtained by the above-mentioned construction method, and judge whether the processed product to be tested is of qualified quality based on the presence or absence of peak 13 and whether it complies with 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 hyacinth bean (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] Refer to the method under "2" to prepare 1 portion of white lentil test solution, accurately pipette 5 μl of 1 portion of white lentil test solution, inject it into the ultra-high performance liquid chromatograph, and determine it according to the chromatographic conditions under "3" to obtain 1 portion of white lentil chromatogram, see Fig.11 .

[0223] contrast Fig.11 and Figure 2 (The white lentil reference characteristic spectrum of Example 1) It can be seen that the number of chromatographic peaks in the chromatogram obtained using the ultraviolet detector is small, and the reason may be that some components have a low response or no response under the ultraviolet detector conditions, which cannot well reflect the overall quality of 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 hyacinth bean (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 6.0g of the powder of this product (BBD12, passed through No. 3 sieve), accurately weigh it, put it in a stoppered conical flask, add 50mL of 75% ethanol, ultrasonically treat (power 300W, frequency 40kHz) for 30 minutes, cool, filter, and obtain reaction residue and extract. Wash the container and reaction residue with a small amount of 75% ethanol in batches, add the washing liquid into the extract and concentrate to dryness to obtain the extract. Add 3mL of water to dissolve it, pass it through a hydrophilic-lipophilic balance solid phase extraction column (1g, 6mL, pre-washed with 12ml of methanol first, and then pre-washed with 12mL of water), elute with 10mL of 60% methanol, discard the eluent, and then elute with 9mL of methanol into a 10mL 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] Refer to the method under "2" to prepare 1 portion of white lentil test solution, accurately pipette 5 μl of 1 portion of white lentil test solution, inject it into the ultra-high performance liquid chromatograph, and determine it according to the chromatographic conditions under "3" to obtain 1 portion of white lentil chromatogram, see Fig.12 .

[0234] contrast Fig.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 hyacinth bean (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 the test solution See the preparation of the 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] Refer to the method under "2" to prepare 1 portion of white lentil test solution, accurately pipette 5 μl of 1 portion of white lentil test solution, inject it into the ultra-high performance liquid chromatograph, and determine it according to the chromatographic conditions under "3" to obtain 1 portion of white lentil chromatogram, see Fig.13 .

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

[0247] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described 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-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A method for constructing a characteristic spectrum of white hyacinth bean 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; 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 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; Wherein, the chromatographic conditions set by the ultra-high performance liquid chromatograph include: 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; 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 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%; 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 volume concentration of acetic acid in the acetic acid aqueous solution is 0.2% to 0.6%.

3. The construction method according to claim 1, characterized in that: Therefore, the chromatographic conditions also include at least one of the following conditions: 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; 2) Flow rate is 0.40 mL to 0.80 mL per minute; 3) Column temperature is 38℃~42℃; 4) The carrier gas flow rate of the evaporative light scattering detector is 2.7 L / min to 3.1 L / min; 5) The drift tube temperature of the evaporative light scattering detector is 102°C to 110°C.

4. The construction method according to any one of claims 1 to 3, characterized in that: The construction method satisfies at least one of the following characteristics: 1) The extraction solvent is ethanol aqueous solution; 2) The extraction process includes a heating reflux process.

5. The construction method according to any one of claims 1 to 3, characterized in that: The processed white lentil product is fried white lentil.

6. A method for distinguishing white lentils from their counterfeits, characterized in that: The following steps are involved: Extract the sample with an extraction solvent to obtain a sample extract, add the extraction solvent to the sample extract, filter, and take the filtrate as a sample solution; Injecting the test product 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 chromatogram of the test product, and comparing the chromatogram of the test product with the characteristic spectrum of 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 3.

7. The identification method according to claim 6, characterized in that: The identification method satisfies at least one of the following characteristics: 1) The extraction solvent is ethanol aqueous solution; 2) The extraction process includes a heating reflux process.

8. The identification method according to any one of claims 6 to 7, characterized in that: The fake white hyacinth bean is at least one of white kidney bean, lentil bean, white eyebrow bean and white sword bean.

9. A method for detecting processed white hyacinth bean, characterized in that: The following steps are involved: Extract the processed product to be tested with an extraction solvent to obtain an extract of the processed product to be tested, add the extraction solvent to the extract of the processed product to be tested, filter, and take the filtrate as a solution of the processed product to be tested; 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, 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 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 3.

10. The detection method according to claim 9, characterized in that: The detection method meets at least one of the following characteristics: 1) The extraction solvent is ethanol aqueous solution; 2) The extraction process includes a heating reflux process.

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