A soybean powder identification method based on lipidomics

Abstract

The present invention relates to a soybean powder identification method based on lipidomics. The identification method detects different types of soybean powder samples based on non-targeted lipidomics technology combined with ultra-high performance liquid chromatography-time-of-flight mass spectrometry, obtains lipidomics data of soybean powders of different types, semi-quantitatively analyzes lipid components in combination with standard products, and then uses chemometric methods to screen characteristic differential lipid components of soybean powders of different types, which are then used for soybean powder identification. The method of the present invention can accurately identify the type of soybean powder and prevent soybean powder counterfeiting.

Description

Technical Field

[0001] The invention relates to the field of food detection, and in particular to a method for identifying soybean powder. Background Art

[0002] Legumes refer to the general term for the legumes in the dicotyledonous plants, so they are generally called legumes, also known as beans. There are many types of beans, such as soybeans, mung beans, chickpeas, peas, broad beans, cowpeas, red beans, kidney beans, etc. Beans are rich in nutrients such as dietary protein, essential amino acids, dietary fiber, vitamins and minerals, and are an important part of a healthy diet. In addition, beans also contain a variety of bioactive ingredients and have a variety of nutritional benefits. For example, the isoflavones in soybeans can enhance the body's physical fitness and disease resistance, and have the effects of lowering blood pressure and losing weight; mung beans have the effects of clearing heat and detoxifying, reducing swelling, etc.; chickpeas are rich in isoflavones, chickpea sprouts and other active ingredients, which have the effects of lowering blood sugar, treating bronchitis, and preventing diabetes; kidney beans contain saponins, urease and other active ingredients, which have the effects of improving the body's own immunity and inhibiting the development of tumor cells. There are obvious differences in the quality and nutritional composition of various types of beans. Conducting research on soybean powder identification is particularly important for understanding the authenticity of commercially available soybean powder. This can prevent counterfeit and inferior products from entering the market and provide consumers with high-quality soybean powder products.

[0003] Non-targeted lipidomics technology can analyze thousands of known and unknown lipid components at the same time. It has a complete database that includes spectra of more than 8 major categories, 300 subcategories, and 1.7 million lipid molecules. In recent years, more and more studies have used this technology to conduct authenticity identification research on food and agricultural products.

[0004] At present, there is no report on soybean powder identification based on non-targeted lipidomics technology. In order to fill the technical gap in this field, it is necessary to establish a soybean powder identification based on lipidomics. Summary of the invention

[0005] The object of the present invention is to provide a bean powder identification method based on characteristic differential lipid components. The method is based on non-targeted lipidomics analysis technology, utilizes ultra-high performance liquid chromatography-time-of-flight mass spectrometry to collect lipid component data of different types of bean powder, and screens the characteristic differential lipid components of each type of bean powder through chemometrics to distinguish the bean powder of each type of bean.

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions:

[0007] The first aspect of the present invention provides a soybean powder identification method based on characteristic difference lipid components, comprising the following steps:

[0008] (1) collecting different types of bean samples, removing impurities from each sample, grinding them into powder, adding an extracting solution, extracting by vortex oscillation and then centrifuging, collecting the supernatant, drying with nitrogen gas, adding a re-dissolving solution, re-dissolving and filtering to obtain a filtrate of the sample to be tested;

[0009] (2) performing non-targeted lipidomics analysis on the sample filtrate of different types of bean powder obtained in step (1) using ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry to obtain lipid data of the different types of bean powder samples, and processing the lipid data using mass spectrometry data analysis software;

[0010] (3) All identified lipid components were qualitatively identified using the LIPID MAPS database, and the software was used for molecular prediction to determine the structure of the screened lipid components. The lipid components were then semi-quantitatively analyzed using lipid isotope internal standards.

[0011] (4) Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to analyze the lipid data of different types of bean flour samples and an OPLS-DA model was constructed. Based on the VIP value of the OPLS-DA model and the p-value of ANOVA, the characteristic differential lipid components of different types of bean flour were screened out.

[0012] Preferably, the bean sample in step (1) is selected from kidney beans, broad beans, black soybeans, red beans, cowpeas, mung beans, peas or chickpeas.

[0013] Preferably, the extracting solution used in step (1) is a mixed solvent of methanol and methyl tert-butyl ether in a volume ratio of 1:2.

[0014] Preferably, the reconstitution solution used in step (1) is a mixed solvent of dichloromethane and methanol in a volume ratio of 1:1.

[0015] Preferably, the chromatographic conditions for detection by ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry in step (2) are as follows: the chromatographic column is a Phenomenex Kinetex C18 chromatographic column; the mobile phase A is a mixed solution of water, methanol and acetonitrile in a volume ratio of 1:1:1 containing 5 mM ammonium acetate, and the mobile phase B is a mixed solution of isopropanol and acetonitrile in a volume ratio of 5:1 containing 5 mM ammonium acetate; the flow rate is 0.4 mL / min; and the gradient elution program is as follows:

[0016] time Mobile phase A Mobile phase B 0-0.5min 80％ 20％ 0.5-1.5min 80％-60％ 20％-40％ 1.5-3.0min 60％-40％ 40％-60％ 3.0-13min 40％-2％ 60％-98％ 13.0-13.1min 2％-80％ 98％-20％ 13.1-17.0min 80％ 20％ .

[0017] Preferably, the mass spectrometry conditions for detection by ultra-high performance liquid chromatography tandem time-of-flight mass spectrometer in step (2) are as follows: using an electrospray ionization ion source ESI, collecting data in positive ion and negative ion modes respectively, the ion source temperature is 600°C, the collision energy is 35eV, the positive ion mode spray voltage is +5500V, and the negative ion mode is -4500V.

[0018] Preferably, in step (3), MS-DIAL, MS-FINDER and PeakView software are used for molecular prediction.

[0019] Preferably, in step (4), based on the VIP value of the OPLS-DA model and the p value of ANOVA, metabolites with VIP>1 and p<0.05 are selected as characteristic differential lipid components for distinguishing different soybean powders.

[0020] More preferably, in step (4), metabolites with VIP>1.1 and p<0.05 are selected as characteristic differential lipid components for distinguishing different soybean powders.

[0021] Further preferably, in step (4), metabolites with VIP>1.2 and p<0.05 are selected as characteristic differential lipid components for distinguishing different soy flours.

[0022] Preferably, the characteristic difference lipid components screened out in step (4) are selected from DGGA 34:2, DGGA 34:3, DGGA 36:2, DGGA 36:3, DGGA 36:4, DGGA 36:6, HexCer 31:3; O2, HexCer 32:1; O4, HexCer32:2; O3, HexCer 32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PA36:2, PC 30:2, PC 32:2, PC 33:2, PC 36:0, PC 37:1, PC 37:2, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PC 41:1, PE32:2, PE 34:2, PE 36:2, PE 37:1, PE 37:2, PE 37:3, PE 37:4, PE 38:1, PE 38:2, PE 38:3, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE 42:2, PG 31:1, PG 34:0, PI 32:0, PI 32:3, PI33:1, PI 33:2, PI 39:2, PI 40:2, PI 41:2, PI 41:3, SM 34:1; O2, SM 37:6;O2,MGDG 36:2,MGDG 36:3,MGDG 36:5,DGDG 35:2,DGDG 36:3,DGDG 36:4,DG 33:1,DG 36:0,DG 36:2,DG36:3,DG 38:0,DG 40:0,DG 40:1,DG 42:0,DG 42:3,DG 44:2,TG 46:0,TG 46:1,TG 46:2,TG 46:3,TG 47:2,TG 48:1,TG 48:4,TG 49:0,TG 49:1,TG 49:2,TG 49:3,TG 50:3,TG50:6,TG 51:3,TG 51:4,TG 51:6,TG 51:7,TG 52:4,TG 53:5,TG 53:6,TG 54:0,TG 54:6,TG 55:2,TG 55:3,TG 55:5,TG 55:6,TG 55:7,TG 56:1,TG 56:2,TG 56:4,TG 56:5,TG56:6,TG 56:7,TG 57:2,TG 57:4,TG 58:1,TG 58:2,TG 58:3,TG 58:5,TG 58:6,TG 58:7,TG 59:2,TG 59:3,TG 59:5,TG 60:1,TG 60:2,TG 60:3,TG 60:4,TG 60:7,TG 61:0,TG61:4,TG 61:6,TG 63:4,TG 64:1,TG 64:2,TG 64:3,TG 64:6,TG 65:3,TG 66:2,TG 66:3,TG 66:4,TG 72:4,TG 72:5,TG 74:5,Cer 31:1;O3,Cer 32:2;O3,Cer 32:3;O2,Cer 33:1;O3,Cer 34:0;O3,Cer 34:1;O3,Cer 34:1;O4,Cer 34:2;O2,Cer 36:0;O3,Cer 36:1;O3,Cer 39:0;O3,Cer 40:0;O4,Cer 40:1;O2,Cer 40:1;O3,Cer 40:1;O4,Cer 40:2;O2,Cer41:0;O4,Cer 41:1;O3,Cer 41:1;O4,Cer 41:2;O2,Cer 42:0;O3,Cer 42:1;O2,Cer 42:2;O2,Cer 43:0;O3,Cer 43:0;O4,Cer 43:3;O4,Cer 44:4;O3, FA25:0, FA28:0, FA29:0, FA34:1, FA 36:0, FA 36:1, FA 36:2, FA 38:0, FA40:0, FA40:1, FA40:2, FA42:1 or more. ;

[0023] Preferably, the characteristic differential lipid components screened out in step (4) are HexCer 34:1; O4, PC 38:3, PE 39:2, PE 41:2, PE 42:2, PG 31:1, PI 41:2, TG 56:6, TG 56:7, Cer 34:0; O3, Cer 34:1; O3, Cer 36:0; O3, Cer 39:0; O3, Cer 40:0; O4, Cer 40:1; O3, Cer 41:0; O4, Cer 43:0; O4, Cer 43:3; O4, a combination of FA28:0 and FA29:0.

[0024] Preferably, the characteristic differential lipid components screened out in step (4) are DGGA 36:6, HexCer 32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PC 32:2, PC 33:2, PC 37:1, PC 38:1, PC 38:2, PC38:3, PC 40:2, PE 34:2, PE 37:1, PE 37:2, PE 37:4, PE 38:1, PE 38:2, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE 42:2, PG 31:1, PI 41:2, MGDG 36:2, MGDG 36:5, DG 40:0, DG 40:1, DG 42:0, DG 42:3, TG 48:4, TG 50:3, TG 53:6, TG 55:7, TG 56:4, TG 56:5, TG 56:6, TG56:7, TG 57:2, TG 58:1, TG 58:5, TG 60:4, TG 60:7, TG 64:2, TG 66:2, TG 66:3, TG 72:4, TG 72:5, Cer 34:0; O3, Cer 34:1; O3, Cer 34:1; O4, Cer 36:0; O3, Cer39:0; O3, Cer 40:0; O4, Cer 40:1; O2, Cer 40:1; 40:1; O4, Cer 40:2; O2, Cer 41:0; O4, Cer 41:1; O3, Cer 41:1; O4, Cer 41:2; O2, Cer 42:1; O2, Cer 42:2; O2, Cer 43:0; O3, Cer 43:0; O4, Cer43:3; O4, FA28:0, FA29:0, FA 36:1, a combination of FA40:0 and FA42:1.

[0025] Preferably, the characteristic difference lipid components screened out in step (4) are DGGA 34:2, DGGA 34:3, DGGA 36:2, DGGA 36:3, DGGA 36:4, DGGA 36:6, HexCer 31:3; O2, HexCer 32:1; O4, HexCer32:2; O3, HexCer 32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PA 36:2, PC 30:2, PC 32:2, PC 33:2, PC 36:0, PC 37:1, PC 37:2, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PC 41:1, PE32:2, PE 34:2, PE 36:2, PE 37:1, PE 37:2, PE 37:3, PE 37:4, PE 38:1, PE 38:2, PE 38:3, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE 42:2, PG 31:1, PG 34:0, PI 32:0, PI 32:3, PI33:1, PI 33:2, PI 39:2, PI 40:2, PI 41:2, PI 41:3, SM 34:1; O2, SM 37:6;O2,MGDG 36:2,MGDG 36:3,MGDG 36:5,DGDG 35:2,DGDG 36:3,DGDG 36:4,DG 33:1,DG 36:0,DG 36:2,DG36:3,DG 38:0,DG 40:0,DG 40:1,DG 42:0,DG 42:3,DG 44:2,TG 46:0,TG 46:1,TG 46:2,TG 46:3,TG 47:2,TG 48:1,TG 48:4,TG 49:0,TG 49:1,TG 49:2,TG 49:3,TG 50:3,TG50:6,TG 51:3,TG 51:4,TG 51:6,TG 51:7,TG 52:4,TG 53:5,TG 53:6,TG 54:0,TG 54:6,TG 55:2,TG 55:3,TG 55:5,TG 55:6,TG 55:7,TG 56:1,TG 56:2,TG 56:4,TG 56:5,TG56:6,TG 56:7,TG 57:2,TG 57:4,TG 58:1,TG 58:2,TG 58:3,TG 58:5,TG 58:6,TG 58:7,TG 59:2,TG 59:3,TG 59:5,TG 60:1,TG 60:2,TG 60:3,TG 60:4,TG 60:7,TG 61:0,TG61:4,TG 61:6,TG 63:4,TG 64:1,TG 64:2,TG 64:3,TG 64:6,TG 65:3,TG 66:2,TG 66:3,TG 66:4,TG 72:4,TG 72:5,TG 74:5,Cer 31:1;O3,Cer 32:2;O3,Cer 32:3;O2,Cer 33:1;O3,Cer 34:0;O3,Cer 34:1;O3,Cer 34:1;O4,Cer 34:2;O2,Cer 36:0;O3,Cer 36:1;O3,Cer 39:0;O3,Cer 40:0;O4,Cer 40:1;O2,Cer 40:1;O3,Cer 40:1;O4,Cer 40:2;O2,Cer41:0;O4,Cer 41:1;O3,Cer 41:1;O4,Cer 41:2;O2,Cer 42:0;O3,Cer 42:1;O2,Cer 42:2;O2,Cer 43:0;O3,Cer 43:0;O4,Cer 43:3;O4,Cer 44:4;Combination of O3, FA25:0, FA28:0, FA29:0, FA34:1, FA 36:0, FA 36:1, FA 36:2, FA 38:0, FA40:0, FA40:1, FA40:2 and FA42:1. ;

[0026] Preferably, the names of the characteristic difference lipid components in step (4) are determined according to the lipid classification and naming system in the LIPID MAPS database.

[0027] The second aspect of the present invention provides a composition comprising characteristic differential lipid components for soybean powder identification, wherein the characteristic differential lipid components are selected from DGGA 34:2, DGGA 34:3, DGGA 36:2, DGGA 36:3, DGGA 36:4, DGGA36:6, HexCer 31:3; O2, HexCer 32:1; O4, HexCer 32:2; O3, HexCer 32:3; O2, HexCer34:1; O4, HexCer 44:1; O4, PA 36:2, PC 30:2, PC 32:2, PC 33:2, PC 36:0, PC 37:1, PC 37:2, PC38:1, PC 38:2, PC 38:3, PC 40:2, PC 41:1, PE 32:2, PE 34:2, PE 36:2, PE 37:1, PE 37:2, PE 37:3, PE 37:4, PE 38:1, PE 38:2, PE 38:3, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE42:2, PG 31:1, PG 34:0, PI 32:0, PI 32:3, PI 33:1, PI 33:2, PI 39:2, PI 40:2, PI 41:2, PI 41:3, SM 34:1; O2, SM 37:6;O2,MGDG 36:2,MGDG 36:3,MGDG 36:5,DGDG 35:2,DGDG36:3,DGDG 36:4,DG 33:1,DG 36:0,DG 36:2,DG 36:3,DG 38:0,DG 40:0,DG 40:1,DG 42:0,DG 42:3,DG 44:2,TG 46:0,TG 46:1,TG 46:2,TG 46:3,TG 47:2,TG 48:1,TG 48:4,TG49:0,TG49:1,TG49:2,TG49:3,TG50:3,TG50:6,TG51:3,TG51:4,TG51:6,TG51:7,TG52:4,TG53:5,TG53:6,TG54:0,TG54:6,TG55:2,TG55:3,TG55:5,TG55:6,TG55:7,TG56:1,TG56:2,TG56:4,TG56:5,TG56:6,TG56:7,TG 57:2,TG 57:4,TG 58:1,TG 58:2,TG 58:3,TG 58:5,TG 58:6,TG 58:7,TG 59:2,TG 59:3,TG 59:5,TG 60:1,TG60:2,TG 60:3,TG 60:4,TG 60:7,TG 61:0,TG 61:4,TG 61:6,TG 63:4,TG 64:1,TG 64:2,TG 64:3,TG 64:6,TG 65:3,TG 66:2,TG 66:3,TG 66:4,TG 72:4,TG 72:5,TG 74:5,Cer31:1;O3,Cer 32:2;O3,Cer 32:3;O2,Cer 33:1;O3,Cer 34:0;O3,Cer 34:1;O3,Cer 34:1;O4,Cer 34:2;O2,Cer 36:0;O3,Cer 36:1;O3,Cer 39:0;O3,Cer 40:0;O4,Cer 40:1;O2,Cer 40:1;O3,Cer 40:1;O4,Cer 40:2;O2,Cer 41:0;O4,Cer 41:1;O3,Cer 41:1;O4,Cer41:2;O2,Cer 42:0;O3,Cer 42:1;O2,Cer 42:2;O2,Cer 43:0;O3,Cer 43:0;O4,Cer 43:3;O4,Cer 44:4;O3, two or more of FA25:0, FA28:0, FA29:0, FA 34:1, FA 36:0, FA 36:1, FA 36:2, FA 38:0, FA40:0, FA40:1, FA40:2 and FA42:1. ;

[0028] Preferably, the characteristic differential lipid components are HexCer 34:1; O4, PC 38:3, PE 39:2, PE 41:2, PE 42:2, PG 31:1, PI 41:2, TG 56:6, TG 56:7, Cer 34:0; O3, Cer 34:1; O3, Cer 36:0; O3, Cer 39:0; O3, Cer 40:0; O4, Cer 40:1; O3, Cer 41:0; O4, Cer 43:0; O4, Cer 43:3; O4, a combination of FA28:0 and FA29:0.

[0029] Preferably, the characteristic difference lipid components are DGGA 36:6, HexCer 32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PC 32:2, PC 33:2, PC 37:1, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PE 34:2, PE 37:1, PE 37:2, PE 37:4, PE 38:1, PE 38:2, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE42:2, PG 31:1, PI 41:2, MGDG 36:2, MGDG 36:5, DG 40:0, DG 40:1, DG 42:0, DG 42:3, TG48:4, TG 50:3, TG 53:6, TG 55:7, TG 56:4, TG 56:5, TG 56:6, TG 56:7, TG 57:2, TG 58:1, TG 58:5, TG 60:4, TG 60:7, TG 64:2, TG 66:2, TG 66:3, TG 72:4, TG 72:5, Cer 34:0; O3, Cer 34:1; O3, Cer 34:1; O4, Cer 36:0; O3, Cer 39:0; O3, Cer 40:0; O4, Cer 40:1; O2, Cer40:1; O3, Cer 40:1; O4, Cer 40:2; O2, Cer 41:0; O4, Cer 41:1; O3, Cer 41:1; O4, Cer 41:2; O2, Cer 42:1; O2, Cer 42:2; O2, Cer 43:0; O3, Cer 43:0; O4, Cer 43:3; O4, FA28:0, FA29:0, FA 36:1, FA40:0 and FA42:1.

[0030] Preferably, the characteristic difference lipid components are DGGA 34:2, DGGA 34:3, DGGA 36:2, DGGA 36:3, DGGA 36:4, DGGA 36:6, HexCer 31:3; O2, HexCer 32:1; O4, HexCer 32:2; O3, HexCer32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PA 36:2, PC 30:2, PC 32:2, PC 33:2, PC 36:0, PC37:1, PC 37:2, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PC 41:1, PE 32:2, PE 34:2, PE 36:2, PE 37:1, PE 37:2, PE 37:3, PE 37:4, PE 38:1, PE 38:2, PE 38:3, PE 39:2, PE 40:2, PE41:1, PE 41:2, PE 42:2, PG 31:1, PG 34:0, PI 32:0, PI 32:3, PI 33:1, PI 33:2, PI 39:2, PI 40:2, PI 41:2, PI 41:3, SM 34:1; O2, SM 37:6;O2,MGDG 36:2,MGDG 36:3,MGDG 36:5,DGDG 35:2,DGDG 36:3,DGDG 36:4,DG 33:1,DG 36:0,DG 36:2,DG 36:3,DG 38:0,DG 40:0,DG 40:1,DG 42:0,DG 42:3,DG 44:2,TG 46:0,TG 46:1,TG 46:2,TG 46:3,TG 47:2,TG48:1,TG 48:4,TG 49:0,TG 49:1,TG 49:2,TG 49:3,TG 50:3,TG 50:6,TG 51:3,TG 51:4,TG 51:6,TG 51:7,TG 52:4,TG 53:5,TG 53:6,TG 54:0,TG 54:6,TG 55:2,TG 55:3,TG55:5,TG 55:6,TG 55:7,TG 56:1,TG 56:2,TG 56:4,TG 56:5,TG 56:6,TG 56:7,TG 57:2,TG 57:4,TG 58:1,TG 58:2,TG 58:3,TG 58:5,TG 58:6,TG 58:7,TG 59:2,TG 59:3,TG59:5,TG 60:1,TG 60:2,TG 60:3,TG 60:4,TG 60:7,TG 61:0,TG 61:4,TG 61:6,TG 63:4,TG 64:1,TG 64:2,TG 64:3,TG 64:6,TG 65:3,TG 66:2,TG 66:3,TG 66:4,TG 72:4,TG72:5,TG 74:5,Cer 31:1;O3,Cer 32:2;O3,Cer 32:3;O2,Cer 33:1;O3,Cer 34:0;O3,Cer34:1;O3,Cer 34:1;O4,Cer 34:2;O2,Cer 36:0;O3,Cer 36:1;O3,Cer 39:0;O3,Cer 40:0;O4,Cer 40:1;O2,Cer 40:1;O3,Cer 40:1;O4,Cer 40:2;O2,Cer 41:0;O4,Cer 41:1;O3,Cer 41:1;O4,Cer 41:2;O2,Cer 42:0;O3,Cer 42:1;O2,Cer 42:2;O2,Cer 43:0;O3,Cer43:0;O4,Cer 43:3;O4,Cer 44:4;A combination of O3, FA25:0, FA28:0, FA29:0, FA 34:1, FA 36:0, FA 36:1, FA 36:2, FA 38:0, FA40:0, FA40:1, FA40:2 and FA42:1. ;

[0031] Preferably, the name of the characteristic difference lipid component is determined according to the lipid classification and naming system in the LIPID MAPS database.

[0032] Preferably, the bean flour is bean flour prepared from kidney beans, broad beans, black soybeans, adzuki beans, cowpeas, mung beans, peas or chickpeas.

[0033] The third aspect of the present invention provides an application of a composition comprising a characteristic difference lipid component in soybean powder identification, wherein the characteristic difference lipid component is selected from DGGA 34:2, DGGA 34:3, DGGA 36:2, DGGA 36:3, DGGA 36:4, DGGA 36:6, HexCer 31:3; O2, HexCer 32:1; O4, HexCer 32:2; O3, HexCer 32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PA 36:2, PC 30:2, PC 32:2, PC 33:2, PC 36:0, PC 37:1, PC 37:2, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PC 41:1, PE 32:2, PE 34:2, PE 36:2, PE37:1, PE 37:2, PE 37:3, PE 37:4, PE 38:1, PE 38:2, PE 38:3, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE 42:2, PG 31:1, PG 34:0, PI 32:0, PI 32:3, PI 33:1, PI 33:2, PI 39:2, PI40:2, PI 41:2, PI 41:3, SM 34:1; O2, SM 37:6;O2,MGDG 36:2,MGDG 36:3,MGDG 36:5,DGDG35:2,DGDG 36:3,DGDG 36:4,DG 33:1,DG 36:0,DG 36:2,DG 36:3,DG 38:0,DG 40:0,DG40:1,DG 42:0,DG 42:3,DG 44:2,TG 46:0,TG 46:1,TG 46:2,TG 46:3,TG 47:2,TG 48:1,TG 48:4,TG 49:0,TG 49:1,TG 49:2,TG 49:3,TG 50:3,TG 50:6,TG 51:3,TG 51:4,TG51:6,TG 51:7,TG 52:4,TG 53:5,TG 53:6,TG 54:0,TG 54:6,TG 55:2,TG 55:3,TG 55:5,TG 55:6,TG 55:7,TG 56:1,TG 56:2,TG 56:4,TG 56:5,TG 56:6,TG 56:7,TG 57:2,TG57:4,TG 58:1,TG 58:2,TG 58:3,TG 58:5,TG 58:6,TG 58:7,TG 59:2,TG 59:3,TG 59:5,TG 60:1,TG 60:2,TG 60:3,TG 60:4,TG 60:7,TG 61:0,TG 61:4,TG 61:6,TG 63:4,TG64:1,TG 64:2,TG 64:3,TG 64:6,TG 65:3,TG 66:2,TG 66:3,TG 66:4,TG 72:4,TG 72:5,TG 74:5,Cer 31:1;O3,Cer 32:2;O3,Cer 32:3;O2,Cer 33:1;O3,Cer 34:0;O3,Cer 34:1;O3,Cer 34:1;O4,Cer 34:2;O2,Cer 36:0;O3,Cer 36:1;O3,Cer 39:0;O3,Cer 40:0;O4,Cer 40:1;O2,Cer 40:1;O3,Cer 40:1;O4,Cer 40:2;O2,Cer 41:0;O4,Cer 41:1;O3,Cer41:1;O4,Cer 41:2;O2,Cer 42:0;O3,Cer 42:1;O2,Cer 42:2;O2,Cer 43:0;O3,Cer 43:0;O4,Cer 43:3;O4,Cer 44:4;O3, two or more of FA25:0, FA28:0, FA29:0, FA 34:1, FA 36:0, FA 36:1, FA36:2, FA 38:0, FA40:0, FA40:1, FA40:2 and FA42:1. ;

[0034] Preferably, the characteristic differential lipid components are HexCer 34:1; O4, PC 38:3, PE 39:2, PE 41:2, PE 42:2, PG 31:1, PI 41:2, TG 56:6, TG 56:7, Cer 34:0; O3, Cer 34:1; O3, Cer 36:0; O3, Cer 39:0; O3, Cer 40:0; O4, Cer 40:1; O3, Cer 41:0; O4, Cer 43:0; O4, Cer 43:3; O4, a combination of FA28:0 and FA29:0.

[0035] Preferably, the characteristic difference lipid components are DGGA 36:6, HexCer 32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PC 32:2, PC 33:2, PC 37:1, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PE 34:2, PE 37:1, PE 37:2, PE 37:4, PE 38:1, PE 38:2, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE42:2, PG 31:1, PI 41:2, MGDG 36:2, MGDG 36:5, DG 40:0, DG 40:1, DG 42:0, DG 42:3, TG48:4, TG 50:3, TG 53:6, TG 55:7, TG 56:4, TG 56:5, TG 56:6, TG 56:7, TG 57:2, TG 58:1, TG 58:5, TG 60:4, TG 60:7, TG 64:2, TG 66:2, TG 66:3, TG 72:4, TG 72:5, Cer 34:0; O3, Cer 34:1; O3, Cer 34:1; O4, Cer 36:0; O3, Cer 39:0; O3, Cer 40:0; O4, Cer 40:1; O2, Cer40:1; O3, Cer 40:1; O4, Cer 40:2; O2, Cer 41:0; O4, Cer 41:1; O3, Cer 41:1; O4, Cer 41:2; O2, Cer 42:1; O2, Cer 42:2; O2, Cer 43:0; O3, Cer 43:0; O4, Cer 43:3; O4, FA28:0, FA29:0, FA 36:1, FA40:0 and FA42:1.

[0036] Preferably, the characteristic difference lipid components are DGGA 34:2, DGGA 34:3, DGGA 36:2, DGGA 36:3, DGGA 36:4, DGGA 36:6, HexCer 31:3; O2, HexCer 32:1; O4, HexCer 32:2; O3, HexCer32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PA 36:2, PC 30:2, PC 32:2, PC 33:2, PC 36:0, PC37:1, PC 37:2, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PC 41:1, PE 32:2, PE 34:2, PE 36:2, PE 37:1, PE 37:2, PE 37:3, PE 37:4, PE 38:1, PE 38:2, PE 38:3, PE 39:2, PE 40:2, PE41:1, PE 41:2, PE 42:2, PG 31:1, PG 34:0, PI 32:0, PI 32:3, PI 33:1, PI 33:2, PI 39:2, PI 40:2, PI 41:2, PI 41:3, SM 34:1; O2, SM 37:6;O2,MGDG 36:2,MGDG 36:3,MGDG 36:5,DGDG 35:2,DGDG 36:3,DGDG 36:4,DG 33:1,DG 36:0,DG 36:2,DG 36:3,DG 38:0,DG 40:0,DG 40:1,DG 42:0,DG 42:3,DG 44:2,TG 46:0,TG 46:1,TG 46:2,TG 46:3,TG 47:2,TG48:1,TG 48:4,TG 49:0,TG 49:1,TG 49:2,TG 49:3,TG 50:3,TG 50:6,TG 51:3,TG 51:4,TG 51:6,TG 51:7,TG 52:4,TG 53:5,TG 53:6,TG 54:0,TG 54:6,TG 55:2,TG 55:3,TG55:5,TG 55:6,TG 55:7,TG 56:1,TG 56:2,TG 56:4,TG 56:5,TG 56:6,TG 56:7,TG 57:2,TG 57:4,TG 58:1,TG 58:2,TG 58:3,TG 58:5,TG 58:6,TG 58:7,TG 59:2,TG 59:3,TG59:5,TG 60:1,TG 60:2,TG 60:3,TG 60:4,TG 60:7,TG 61:0,TG 61:4,TG 61:6,TG 63:4,TG 64:1,TG 64:2,TG 64:3,TG 64:6,TG 65:3,TG 66:2,TG 66:3,TG 66:4,TG 72:4,TG72:5,TG 74:5,Cer 31:1;O3,Cer 32:2;O3,Cer 32:3;O2,Cer 33:1;O3,Cer 34:0;O3,Cer34:1;O3,Cer 34:1;O4,Cer 34:2;O2,Cer 36:0;O3,Cer 36:1;O3,Cer 39:0;O3,Cer 40:0;O4,Cer 40:1;O2,Cer 40:1;O3,Cer 40:1;O4,Cer 40:2;O2,Cer 41:0;O4,Cer 41:1;O3,Cer 41:1;O4,Cer 41:2;O2,Cer 42:0;O3,Cer 42:1;O2,Cer 42:2;O2,Cer 43:0;O3,Cer43:0;O4,Cer 43:3;O4,Cer 44:4;A combination of O3, FA25:0, FA28:0, FA29:0, FA 34:1, FA 36:0, FA 36:1, FA 36:2, FA 38:0, FA40:0, FA40:1, FA40:2 and FA42:1. ;

[0037] Preferably, the name of the characteristic difference lipid component is determined according to the lipid classification and naming system in the LIPID MAPS database.

[0038] Preferably, the bean flour is bean flour prepared from kidney beans, broad beans, black soybeans, adzuki beans, cowpeas, mung beans, peas or chickpeas.

[0039] Beneficial effects of the present invention:

[0040] The bean powder identification method based on characteristic difference lipid components provided by the present invention can comprehensively obtain lipid component information of bean powder of different types, and can screen the characteristic difference lipid components of bean powder of different types by orthogonal partial least squares discriminant analysis. The bean powder of different types of beans can be well distinguished based on the characteristic difference lipid components, thereby providing an excellent detection method for the identification of commercially available bean powder. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 OPLS-DA score graph (A) and permutation test graph (B) to distinguish different bean flours, where (AB) represents adzuki bean, (BS) represents black soybean, (C) represents cowpea, (CP) represents chickpea, (FB) represents broad bean, (KB) represents kidney bean, (MB) represents mung bean, and (P) represents pea;

[0042] Figure 2 This is a hierarchical cluster analysis diagram based on characteristic differential lipid components, where (AB) represents adzuki bean, (BS) represents black soybean, (C) represents cowpea, (CP) represents chickpea, (FB) represents broad bean, (KB) represents kidney bean, (MB) represents mung bean, and (P) represents pea. DETAILED DESCRIPTION

[0043] The invention is further explained below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the invention and are not used to limit the scope of the invention. After reading the invention, various equivalent modifications of the invention by those skilled in the art all fall within the scope defined by the claims attached to this application.

[0044] Example 1. Soybean powder identification method based on lipidomics

[0045] 1. Test methods

[0046] Step 1: Collect eight kinds of beans (kidney beans, broad beans, black soybeans, red beans, cowpeas, mung beans, peas, chickpeas), select three different bottles for each bean, a total of 24 samples (the origin, variety and year of various beans are shown in Table 1 below), each sample is washed to remove impurities, and freeze-dried for 72 hours. The freeze-dried bean samples are ground into powder, and the powder is stored in a -20℃ refrigerator for later use.

[0047] Table 1 Varieties of 24 bean samples

[0048]

[0049]

[0050] Accurately weigh 0.5g of mung bean powder samples from different origins, place them in a 15mL centrifuge tube, add 5mL of methanol: methyl tert-butyl ether (1:2, v / v) extract, vortex and shake for 1h, then centrifuge at 5000rpm for 10 minutes at 4℃, collect the supernatant, blow dry with nitrogen, and add 1mL of dichloromethane: methanol (50:50, v / v) extract to re-dissolve. The re-solution was filtered with a 0.22μm hydrophilic polytetrafluoroethylene filter membrane to obtain the filtrate, which was the sample for machine detection. The quality control QC sample was prepared by mixing equal amounts of extracts from all samples.

[0051] Step 2: Ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry was used to collect mass spectrometric information of mung bean samples from different origins. The conditions used were as follows:

[0052] Chromatographic conditions: The chromatographic column was a Phenomenex Kinetex C18 column; the mobile phase A was a mixed solution of water, methanol, and acetonitrile in a volume ratio of 1:1:1 containing 5 mM ammonium acetate, and the mobile phase B was a mixed solution of isopropanol and acetonitrile in a volume ratio of 5:1 containing 5 mM ammonium acetate; the flow rate was 0.4 mL / min; the gradient elution program was as follows:

[0053] time Mobile phase A Mobile phase B 0-0.5min 80％ 20％ 0.5-1.5min 80％-60％ 20％-40％ 1.5-3.0min 60％-40％ 40％-60％ 3.0-13min 40％-2％ 60％-98％ 13.0-13.1min 2％-80％ 98％-20％ 13.1-17.0min 80％ 20％ .

[0054] Mass spectrometry conditions: Electrospray ionization (ESI) was used to collect data in positive and negative ion modes, respectively. The ion source temperature was 600°C, the collision energy was 35 eV, the spray voltage was +5500V in positive ion mode, and -4500V in negative ion mode. The mass spectrometer adopted full scan mode, and an information correlation acquisition method was established in combination with automatic dynamic background subtraction. The MS mass detection range was m / z100-1000, and the MS / MS mass detection range was m / z 50-1200. After every 5 samples were collected, an external accurate mass calibration was performed using the automatic calibration fluid transfer system to ensure the high mass accuracy of the mass spectrometer during data acquisition.

[0055] Data acquisition was performed in real time using Analyst, and MarkerView software was used to perform deconvolution, peak extraction, peak alignment and other data processing on the raw data obtained from the mass spectrometry to obtain a data matrix containing retention time, mass-to-charge ratio and ion abundance information for subsequent chemometric analysis.

[0056] Step 3: Use the LIPID MAPS database to qualitatively identify the identified lipid components by comparing their accurate mass, isotope abundance distribution, and secondary ion fragment information. Use MS-DIAL, MS-FINDER, and PeakView software for molecular prediction. Then, use lipid isotope internal standards to semi-quantitatively analyze lipid components.

[0057] Step 4: Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to analyze the lipid component data of different types of bean flour and construct an OPLS-DA model. According to the VIP value of the OPLS-DA model and the p value of ANOVA, metabolites with VIP>1 and p<0.05 were selected as characteristic lipid components to distinguish different types of bean flour.

[0058] 2. Test results

[0059] Figure 1 The score graph (A) and permutation test graph (B) of OPLS-DA of different types of bean flour. From the score graph, it can be seen that the eight types of bean flour based on all lipid components can be well separated. In the permutation test, R2 and Q2 are lower than the original point on the right, and the intersection of the regression line of Q2 and the y-axis is on the negative semi-axis, indicating that the model is effective and reliable.

[0060] According to the VIP value of the OPLS-DA model and the p value of ANOVA, metabolites with VIP>1 and p<0.05 were selected as characteristic lipid components for distinguishing different types of bean flour. In this example, a total of 170 differential metabolites were screened out (see Table 2, the names of the lipid components were determined according to the lipid classification and naming system in the LIPID MAPS database).

[0061] Table 2 VIP values ​​of lipid components of different types of bean flour

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069] Based on the characteristic differential lipid components obtained by hierarchical clustering analysis, it can be intuitively seen that the bean powders of the eight types of beans can be clearly distinguished ( Figure 2 ). For example, 21 lipid components DGGA 36:6, FA 34:1, FA 29:0, DGGA 34:3, FA 36:2, FA 36:1, PC 40:2, PE 40:2, PI 41:2, FA28:0, Cer 42:2; O2, Cer 44:4; O3, Cer42:0; O3, Cer40:1; O3, Cer 40:2; O2, Cer 41:2; O2, Cer 41:1; O3, PE 39:2, PE 41:2, PE 42:2 and PI 39:2 were significantly higher in adzuki bean flour than those in other beans; 23 lipid components TG 56:4, TG 64:2, Cer 42:1; O2, DG40:0, TG 64:1, TG 50:3, TG 49:0, DG 38:0, TG 54:0, DG 44:2, DG 40:1, TG 55:7, DG 42:3, TG 49:2, TG 66:2, TG 66:3, TG 58:6, TG 58:7, TG 59:5, TG 58:5, PI 32:0, FA 36:0 and TG64:6 were the highest in cowpea; 23 lipid components Cer 40:1; O2, TG 46:0, PC 36:0, PG 34:0, SM 37:6; O2, PG 31:1, TG 60:7, Cer 39:0; O3, Cer 40:0; O4, Cer 43:3; O4, Cer 43:0; O4, TG 56:6, Cer 41:0; O4, TG 56:7, FA 38:0, Cer 43:0; O3, HexCer 44:1; O4, FA40:1, FA40:0, FA42:1, FA40:2, DG 36:0 and Cer 32:2; O3 has the highest content in mung bean.

[0070] The above results show that the method for identifying characteristic differential lipid components of soybean powder based on lipidomics provided by the present invention can be used to identify various types of soybean powder, and the method has the advantages of accurate experimental results and high feasibility. The identification method of the present invention uses non-targeted lipidomics technology to comprehensively obtain lipid component information of various types of soybean powder, and combines chemometrics to screen out characteristic differential lipid components that distinguish soybean powder of various types, which has important reference significance for the authenticity identification of soybean powder adulteration.

[0071] Although specific embodiments of the present invention have been described, it will be appreciated by those skilled in the art that various changes and modifications may be made to the present invention without departing from the scope or spirit of the present invention. Therefore, the present invention is intended to cover all such changes and modifications that fall within the scope of the appended claims and their equivalents.

Claims

1. An application of a composition comprising lipid components with characteristic differences in soybean powder identification, characterized in that: The characteristic differential lipid components are DGGA 34:2, DGGA 34:3, DGGA 36:2, DGGA 36:3, DGGA 36:4, DGGA 36:6, HexCer 31:3; O2, HexCer 32:1; O4, HexCer 32:2; O3, HexCer 32:3; O2, HexCer 34:1; O4, HexCer 44:1; O4, PA 36:2, PC 30:2, PC 32:2, PC 33:2, PC 36:0, PC 37:1, PC 37:2, PC 38:1, PC 38:2, PC 38:3, PC 40:2, PC 41:1, PE 32:2, PE 34:2, PE 36:2, PE 37:1, PE 37:2, PE37:3, PE 37:4, PE 38:1, PE 38:2, PE 38:3, PE 39:2, PE 40:2, PE 41:1, PE 41:2, PE 42:2, PG 31:1, PG 34:0, PI 32:0, PI 32:3, PI 33:1, PI 33:2, PI 39:2, PI 40:2, PI 41:2, PI41:3, SM 34:1;O2, SM 37:6;O2,MGDG 36:2,MGDG 36:3,MGDG 36:5,DGDG 35:2,DGDG 36:3,DGDG 36:4,DG 33:1,DG 36:0,DG 36:2,DG 36:3,DG 38:0,DG 40:0,DG 40:1,DG 42:0,DG42:3,DG 44:2,TG 46:0,TG 46:1,TG 46:2,TG 46:3,TG 47:2,TG 48:1,TG 48:4,TG 49:0,TG 49:1,TG 49:2,TG 49:3,TG 50:3,TG 50:6,TG 51:3,TG 51:4,TG 51:6,TG 51:7,TG52:4,TG 53:5,TG 53:6,TG 54:0,TG 54:6,TG 55:2,TG 55:3,TG 55:5,TG 55:6,TG 55:7,TG 56:1,TG 56:2,TG 56:4,TG 56:5,TG 56:6,TG 56:7,TG 57:2,TG 57:4,TG 58:1,TG58:2,TG 58:3,TG 58:5,TG 58:6,TG 58:7,TG 59:2,TG 59:3,TG 59:5,TG 60:1,TG 60:2,TG 60:3,TG 60:4,TG 60:7,TG 61:0,TG 61:4,TG 61:6,TG 63:4,TG 64:1,TG 64:2,TG64:3,TG 64:6,TG 65:3,TG 66:2,TG 66:3,TG 66:4,TG 72:4,TG 72:5,TG 74:5,Cer 31:1;O3,Cer 32:2;O3,Cer 32:3;O2,Cer 33:1;O3,Cer 34:0;O3,Cer 34:1;O3,Cer 34:1;O4,Cer 34:2;O2,Cer 36:0;O3,Cer 36:1;O3,Cer 39:0;O3,Cer 40:0;O4,Cer 40:1;O2,Cer40:1;O3,Cer 40:1;O4,Cer 40:2;O2,Cer 41:0;O4,Cer 41:1;O3,Cer 41:1;O4,Cer 41:2;O2,Cer 42:0;O3,Cer 42:1;O2,Cer 42:2;O2,Cer 43:0;O3,Cer 43:0;O4,Cer 43:3;O4,Cer 44:4;<h2 style=";text-align:left;direction:ltr">O3, FA 25:0, FA 28:0, FA 29:0, FA 34:1, FA 36:0, FA 36:1, FA 36:2, FA 38:0, FA 40:0, FA 40:1, FA 40:2 and FA 42:1 are also available; The bean flour is bean flour prepared from kidney beans, broad beans, black soybeans, red beans, cowpeas, mung beans, peas or chickpeas.

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