Use of mangiferin as a broad-spectrum inhibitor of active carbonyl compounds

Abstract

The application discloses application of mangiferin as a broad-spectrum inhibitor of active carbonyl compounds, and provides application of mangiferin (MGF) as an acrolein (ACR), methyl glyoxal (MGO) and glyoxal (GO) inhibitor, wherein, compared with the prior art, the application discloses a new application of mangiferin or a one-addition product of the mangiferin and the active carbonyl compound, that is, the mangiferin or the one-addition product can effectively capture ACR, MGO and GO to control the content of the ACR, MGO and GO and avoid the ACR, MGO and GO from reacting with nucleophilic biological macromolecules to form various harmful addition or cross-linking products. The mangiferin or the one-addition product of the mangiferin and the active carbonyl compound can be used as a scavenger of ACR, MGO and GO, so that the ACR, MGO and GO generated in the body and in a food processing process are inhibited, and thus the various harmful addition or cross-linking products formed by the reaction of RCS with the nucleophilic biological macromolecules are blocked to cause harm to the human body.

Description

TECHNICAL FIELD

[0001] The present application belongs to biological inhibitors, and particularly relates to the application of mangiferin as a broad-spectrum inhibitor of active carbonyl compounds. BACKGROUND

[0002] Active carbonyl species (RCS) is a class of chemicals with one or more carbonyl groups. Structurally, RCS can be divided into two categories: (1) α, β-unsaturated aldehyde containing carbonyl and vinyl groups, such as acrolein (ACR) and crotonaldehyde (CRO); (2) α-dicarbonyl compounds containing two carbonyl groups, such as methylglyoxal (MGO) and glyoxal (GO). Due to the high electron density of the carbonyl carbon atom, RCS is a strong electrophile, so it can easily induce abnormal modification of DNA and protein, causing cell apoptosis, inflammation and tissue damage, and thus become an inducer of aging, obesity and some chronic diseases.

[0003] Acrolein (ACR) is the simplest α, β-unsaturated carbonyl compound with the chemical formula C3H4O. The list of carcinogens published by the International Agency for Research on Cancer of the World Health Organization lists acrolein in the list of category 3 carcinogens. Acrolein contains two electrophilic groups and has strong reactivity, so it can directly adduct with various biomolecules in the body. Water-soluble, unsaturated acrolein can form adducts with glutathione, DNA, protein, etc., accumulate oxygen free radicals, and cause different degrees of harm to the respiratory system, cardiovascular system, reproductive system, nervous system, etc. Therefore, effectively reducing the level of ACR in the body is an effective measure to improve the health status of the human body and thus prevent the occurrence of chronic diseases.

[0004] The sources of ACR are mainly exogenous and endogenous. Exogenous ACR mainly comes from processed foods and beverages with high fat and sugar content. The caramelization reaction, Maillard reaction, oxidation of oil during food processing or long-term storage, and microbial metabolites in fermented foods can all lead to an increase in the content of ACR in food. ACR also exists in the natural environment, and its main sources include herbicides, incomplete combustion of organic matter, kitchen fumes, cigarette smoke, etc. In addition, ACR is also found in environmental water samples such as rivers and rainwater, which mainly come from herbicides, pesticides, drilling water for processing petrochemical industry, etc. ACR in the air accounts for about 11% to 13% of the total atmospheric aldehyde. The ACR in the body mainly comes from lipid peroxidation, Strecker degradation of free amino acids, and oxidation of spermine. Therefore, reducing the formation of ACR in the body and during food processing and storage can simultaneously reduce endogenous and exogenous exposure, improve food safety, and prevent chronic diseases caused by poor diet, which has very important practical significance and theoretical value.

[0005] Glyoxal (GO) and methylglyoxal (MGO) are both 1,2-dicarbonyl reactive compounds, which are highly active glycation factors and have 200-50000 times higher reactivity than glucose. They are precursors of advanced glycation end products (AGEs). AGEs are mainly a class of stable polymers formed by Maillard reaction pathway and other ways between glucose and proteins or amino acids without enzyme participation. The results of researches have proved that the formation and accumulation of AGEs in organs, tissues and plasma of the body can cause chronic diseases such as diabetic complications, cardiovascular and cerebrovascular diseases and senile dementia. Therefore, 1,2-dicarbonyl compounds are taken as the targets for studying protein glycation, and inhibiting the formation of 1,2-dicarbonyl compounds is an effective measure to prevent the occurrence of chronic diseases by inhibiting the formation of AGEs.

[0006] The sources of GO and MGO are mainly divided into exogenous and endogenous. The sources of exogenous GO and MGO include processed foods, automobile exhaust, household cooking and cigarette smoke, and the main sources are some processed foods and beverages with high protein and sugar content. The caramelization reaction, Maillard reaction, oil oxidation of food during heat processing or long-term storage, and microbial metabolites in fermented foods can all lead to the increase of 1,2-dicarbonyl compounds in food. GO and MGO in vivo mainly come from protein glycation, degradation of phosphotriose, sugar degradation of cells, and ketose metabolism of threonine degradation pathways. Therefore, reducing the formation of 1,2-dicarbonyl compounds, blocking the pathway of AGEs formation in vivo and food processing, thereby reducing the intake of exogenous 1,2-dicarbonyl compounds and AGEs, improving food safety, and preventing chronic diseases caused by unhealthy diet have very important practical significance and theoretical value.

[0007] The AGEs inhibitors found in the study mainly include polyphenols and flavonoids. According to the literature "Scavenging of Toxic Acrolein by Resveratrol and Hesperetin and Identification of Adducts", "Inhibitory effect of polyphenols in Houttuynia cordata on advanced glycation end-products (AGEs) by trapping methylglyoxal", "Impact of resveratrol, epicatechin and rosmarinic acid on fluorescent AGEs and cytotoxicity of cookies", it is reported that the ACR inhibition activity of the following substances is strong: hesperidin, morin, resveratrol glycoside, and the inhibition rates are 66%, 75% and 70% respectively; the MGO inhibition activity of the following substances is strong: apigenin, caffeic acid and quercitrin, and the inhibition rates are 49.1%, 50.4% and 47.7% respectively; the GO inhibition activity of the following substances is strong: resveratrol, caffeic acid and kaempferol, and the inhibition rates are 53.3%, 41.2% and 42.6% respectively. However, the inhibition effect of these substances on RCS is not good, and they cannot simultaneously and efficiently inhibit ACR, MGO and GO. In order to effectively make up for this gap, it is necessary to develop new broad-spectrum inhibitors with good ACR, MGO and GO inhibition activities.

[0008] Mangiferin is a highly condensed aromatic ring system, and mangiferin has strong antioxidant activity and various pharmacological effects, such as neuroprotection, anti-diabetes, anti-inflammatory, antipyretic, analgesic, antibacterial, antiviral, anti-tumor and immune regulation. The content of mangiferin in different plants is different, such as mango peel (0.7%), coffee leaves (0.2%) and Anemarrhena (0.41%). Mangiferin is rich in almost every part of mango trees, and it is the main source of mangiferin, and the content in its leaves and peels is the highest. SUMMARY

[0009] The present application provides the application of mangiferin as a broad-spectrum inhibitor of active carbonyl compounds, which can effectively inhibit the production of acrolein in the environment and during food processing and storage, reduce endogenous and exogenous exposure, improve food safety, and simultaneously inhibit the production of methylglyoxal and glyoxal in the environment and organisms, and block the formation of advanced glycation end products, which has very important practical significance and theoretical value for improving environmental protection, food safety and preventing chronic diseases caused by poor diet.

[0010] Technical solution: In order to achieve the above-mentioned purpose, the application of mangiferin in inhibiting active carbonyl compounds, including propylene aldehyde, methyl glyoxal and glyoxal.

[0011] The mangiferin is a natural polyphenol xanthone, and its chemical structural formula is as follows:

[0012]

[0013] The application of the mangiferin in preparing an inhibitor for inhibiting active carbonyl compounds in the environment.

[0014] The application of the mangiferin in preparing an inhibitor for inhibiting active carbonyl compounds generated in food processing.

[0015] The application of the mangiferin in preparing an inhibitor for inhibiting active carbonyl compounds in a living organism.

[0016] As a preferred, the application of the mangiferin in preparing an inhibitor for inhibiting propylene aldehyde, methyl glyoxal and glyoxal in human body, which can prevent the occurrence of chronic diseases of human body, such as tumor, Alzheimer's disease, aging, atherosclerosis or inflammation caused by ACR induced biological nucleophilic macromolecule addition or crosslinking.

[0017] The application of the mangiferin in preparing an inhibitor for inhibiting propylene aldehyde, methyl glyoxal and glyoxal in the environment, food processing or living organism, wherein the mangiferin can inhibit various harmful addition or crosslinking products formed by the reaction of active carbonyl compounds and nucleophilic biological macromolecules.

[0018] The application of the mangiferin in preparing an inhibitor for inhibiting propylene aldehyde, methyl glyoxal and glyoxal in the environment, food processing or living organism, wherein the mangiferin can inhibit various harmful addition or crosslinking products formed by the reaction of active carbonyl compounds and nucleophilic biological macromolecules.

[0019] The addition product includes a addition product MGF-ACR of the mangiferin and propylene aldehyde, and a addition product MGF-MGO of the mangiferin and methyl glyoxal, and the structures are as follows:

[0020]

[0021] The application of the mangiferin in preparing a broad-spectrum inhibitor for carbonyl compounds, wherein the mangiferin is the only component or is used as a main component and is compounded with other substances to form a preparation.

[0022] The application of the mangiferin in preparing a broad-spectrum inhibitor for carbonyl compounds, wherein the mangiferin is the only component or is used as a main component and is compounded with other substances to form a preparation.

[0023] The application of the adduct of mangiferin and active carbonyl compound in inhibiting active carbonyl compound, wherein the active carbonyl compound includes propylene aldehyde, methyl glyoxal and glyoxal, and the adduct is the adduct of mangiferin and propylene aldehyde, MGF-ACR, and the structure is shown as follows:

[0024]

[0025] Or the adduct is the adduct of mangiferin and methyl glyoxal, MGF-MGO, and the structure is shown as follows:

[0026]

[0027] Further, the application of the adduct of mangiferin and active carbonyl compound in preparing an inhibitor of active carbonyl compound in the environment, food processing or organism.

[0028] The inhibitor of active carbonyl compound in the environment, food processing or organism, wherein the inhibitor is composed of mangiferin and / or the adduct of mangiferin and active carbonyl compound as the only component or as the main component in combination with other substances.

[0029] The application of the mango peel and / or mango leaf in preparing an inhibitor of active carbonyl compound, wherein the main component of the mango peel and / or mango leaf is mangiferin, and the active carbonyl compound includes propylene aldehyde, methyl glyoxal and glyoxal.

[0030] In the application, mangiferin (MGF) is a carbon ketone glycoside of tetrahydroxy pyrone, belonging to biflavonoid flavonoid compounds. The application is directed to propylene aldehyde, methyl glyoxal and glyoxal with high toxicity in the environment, food processing or organism, and the adducts are formed after the capture of MGF, and the reaction mechanism of MGF and ACR, MGO and GO is Michael addition reaction, that is, the hydroxyl group on the A ring of MGF structure reacts with the aldehyde group of ACR to form hemiacetal structure after the addition reaction of MGF and the above-mentioned substances under alkaline conditions. The application captures ACR, MGO and GO through MGF, and the broad-spectrum capture activity is good. The application effectively fills the blank of biflavonoid compounds as inhibitors of RCS compounds.

[0031] The general flavonoid structure is a single flavone, such as apigenin, resveratrol, quercitrin; and the mango glycoside as a biflavone has its particularity in structure, and the inhibition effect of ACR, MGO and GO is first proposed in the application, and the broad-spectrum inhibition effect is good. Although many substances in the prior art have certain inhibitory effect on ACR, MGO or GO, such as hesperidin, quercetin, morin, resveratrol glycoside and the like, the structures of these substances are obviously different from the structure of the mango glycoside (MGF) of the application, for example, the parent nucleus structures of quercitrin and mango glycoside are obviously different; and there is no report of a substance that can simultaneously and efficiently degrade ACR, MGO and GO in the prior art. The application of mango glycoside in efficient and broad-spectrum inhibition of ACR, MGO and GO is first proposed, and it is first found that when ACR, MGO and GO coexist, the inhibition effect of mango glycoside on MGO and GO which are difficult to inhibit and degrade is significant.

[0032] In addition, it is found through inhibition experiments that the adduct of mango glycoside and active carbonyl compounds, such as the adduct of mango glycoside and acrolein MGF-ACR, has no significant difference in inhibition effect on ACR compared with MGF of the same concentration, still maintains a high removal efficiency, but can significantly improve the inhibition effect on MGO and GO, and is obviously superior to MGF.

[0033] Beneficial effects: compared with the prior art, the application has the following advantages:

[0034] The application proposes a new application of mango glycoside, that is, it is first proposed that the mango glycoside can be used as a broad-spectrum active carbonyl compound inhibitor to simultaneously inhibit the contents of ACR, MGO and GO, and avoid the formation of various irreversible harmful adducts or crosslinking products by further reacting with nucleophilic biological macromolecules.

[0035] The mango glycoside can be used as an RCS inhibitor to remove ACR, MGO and GO generated in the environment such as water, in the body and in the food processing process, and further block the formation of harmful crosslinking products induced by RCS to prevent the harm to the human body. The inhibition effect of the mango glycoside on ACR and GO can reach more than 80%, and the inhibition effect on MGO can reach more than 70%, and the mango glycoside of the application can simultaneously inhibit ACR, MGO and GO at a low dose, and the effect is significant.

[0036] The application synthesizes a new structure of the adduct of mango glycoside and RCS, and the adduct can also be used as a broad-spectrum active carbonyl compound inhibitor and has a significant effect. BRIEF DESCRIPTION OF DRAWINGS

[0037] Figure 1Reaction pathway mechanism diagram of mangiferin with ACR, MGO and GO (group A is the mechanism diagram of mangiferin with ACR, group B is the mechanism diagram of mangiferin with MGO, and group C is the mechanism diagram of mangiferin with GO)

[0038] Figure 2 Mass spectrum of the adduct of mangiferin with ACR, MGO and GO

[0039] Figure 3 Inhibition activity of different concentrations of mangiferin on three active carbonyl compounds (A is ACR, B is MGO, and C is GO) under simulated processing conditions, each value is the average value ± standard deviation; Tukey's method was used for significance test, different capital letters represent significant differences between different concentration points, and different small letters represent differences between different time points (p<0.05), and the concentration increases from left to right in the column chart;

[0040] Figure 4 Inhibition activity of different concentrations of mangiferin (A is 0.5 mmol / L, B is 1.0 mmol / L, and C is 1.5 mmol / L) on three active carbonyl compounds under simulated processing conditions (the column chart from left to right is ACR, MGO and GO), each value is the average value ± standard deviation; Tukey's method was used for significance test, different capital letters represent significant differences between different RCS, and different small letters represent differences between different time points (p<0.05), and ACR, MGO and GO are arranged from left to right in the column chart;

[0041] Figure 5 Reaction pathway mechanism diagram of mangiferin and RCS under simulated processing conditions, wherein a, b and c diagrams are respectively the reaction pathway diagrams of MGF with ACR-MGO-GO at 1:1, 1:3 and 1:10;

[0042] Figure 6 Reaction of mangiferin and RCS under simulated processing conditions, wherein a and b diagrams are respectively the first mass spectrum and the second mass spectrum of the new adduct MGF-ACR-MGO; and c and d diagrams are respectively the first mass spectrum and the second mass spectrum of the new adduct MGF-2MGO-GO;

[0043] Figure 7The inhibitory activity of the adduct of mangiferin and ACR (MGF-ACR) of the application on ACR (A), MGO (B) and GO (C) was compared under simulated processing for 30 min, and each value was an average value ± standard deviation; Tukey's method was used for significance test, and different capital letters represented significant differences between different inhibitors (p<0.05), and the column chart from left to right was MGF and MGF-GO;

[0044] Figure 8 The results of the activity determination of the mango peel (A) and mangiferin (B) in inhibiting RCS in heavy oil cakes are shown, each value is an average value ± standard deviation; Tukey's method was used for significance test, and different capital letters represented significant differences between different adding amounts (p<0.05), and the concentration in the column chart increased from left to right or the mass increased in turn. DETAILED DESCRIPTION

[0045] In the experimental methods described in the examples, if no special instructions are given, they are all conventional methods; if no special instructions are given, the reagents and materials can be obtained from commercial channels.

[0046] Example 1

[0047] Purification and structure research of the adduct of mangiferin and ACR

[0048] (1) Experimental materials and instruments

[0049] Mangiferin (chromatographically pure, 95%, Shanghai Yuanye Biotechnology Co., Ltd.), propylene aldehyde (98% aqueous solution, Shandong Shiyachemical Industry Co., Ltd.), methanol (analytically pure, Shanghai Reagent Company), pure water (Hangzhou Wahaha Group Co., Ltd.), ODS filler (ODS-A-HG, 50 μm, Japan Fuji Company).

[0050] AVANCE 500MHz nuclear magnetic resonance instrument (Bruker Company); Waters Xevo TQ-XS triple quadrupole mass spectrometer (Waters Company, USA).

[0051] (2) Experimental steps

[0052] ① Preparation of the adduct of mangiferin and ACR

[0053] Dissolve 1.0 g of mangiferin in 5 mL of methanol, add 2 mL of DMSO as a cosolvent, and then add 6 mL of ACR solution diluted with PBS (pH 7.0, 0.1 mol / L) to make the molar ratio of mangiferin to ACR 1:10. React at 100°C for 15 min. After the reaction, add 10 μL of glacial acetic acid, and then ice-bath. After ice-bath, spin-evaporate the reaction solution to about 1 mL. Use glass column chromatography (2.6 cm x 30.0 cm) with ODS silica gel (ODS-A-HG, 50 μm) as the filler to preliminarily separate the adduct. First, remove DMSO in the reaction solution with 10% methanol-water (v / v), and then increase the gradient to 25%. Collect the eluate, and then freeze-dry to obtain the adduct of mangiferin and ACR (MGF-ACR, 102 mg).

[0054] 3. Structure identification of MGF-ACR

[0055] Dissolve MGF-ACR in methanol to prepare a standard solution of 200 ng / mL. Analyze the molecular weight by UPLC-MS / MS, and analyze the structure by 1D-NMR (H, 1 H, 13 C) and 2D-NMR (HMQC, HMBC).

[0056] Mass spectrometry conditions

[0057] Ion source: Electron Spray Ionization (ESI); scanning mode: negative ion scanning; detection mode: Multiple Reaction Monitoring (MRM); capillary voltage: 1.5 kv; desolvation gas temperature: 500°C; desolvation gas flow rate: 1000 L / h; cone gas flow rate: 150 L / h; atomizer flow rate: 7.0 bar; collision gas flow rate: 0.15 mL / min; cone voltage: 25 V. Data acquisition and analysis software: MassLynx 4.2.

[0058] Liquid chromatography conditions

[0059] Instrument: Waters Xevo TQ-XS;

[0060] Chromatographic column: ACQUITY UPLC BEH C18 (2.1 x 100 mm, i.d., 1.7 μm);

[0061] Column temperature: 40°C; flow rate: 0.3 mL / min; injection volume: 2 μL;

[0062] Eluent: 0.1% formic acid aqueous solution (mobile phase A); methanol (mobile phase B);

[0063] Elution gradient: 0-1.5 min, 5% B; 1.5-2 min, 5-15% B; 2-4 min, 15-30% B; 4-6 min, 30-40% B; 6-7 min, 40-70% B; 7-9 min, 70-95% B, 9-10 min, 95% B, 9-11 min, 95-5% B, 11-12 min, 5% B.

[0064] (3) Experimental results

[0065] ① Identification of the MGF-ACR structure

[0066] The prepared MGF-ACR was determined by LC-MS / MS. Figure 2 As shown in the figure, in negative ion mode, the parent ion mass is m / z:477[MH]+. 1 H and 13 C NMR data are shown in Table 1.

[0067] Table 1 Mangiferin and its adducts with ACR and MGO (MGF-ACR, MGF-MGO) 1 H NMR (500 Hz) and 13 CNMR

[0068]

[0069]

[0070] Table 1 shows that MGF-ACR shares a similar carbon skeleton with mangiferin. First, three new carbon signals (δC 93.3, δC 26.4, and δC 15.0) and a chemical shift at position 3 to δC 157.9, combined with the molecular formula, confirm that ACR shares an oxygen atom with C-3, indicating that ACR forms a six-membered ring with the carbon atoms C-3 and C-4 on the benzene ring. Furthermore, a correlation between δH 4.61 (H-1') and δC 107.6 (C-2) indicates that the sugar ring is attached to C-2. Combined with the related signals from H14 to C-3, C-4, and C-16 in the HMBC spectrum, these features ultimately confirmed the structure of MGF-ACR as a new mangiferin derivative. Its structural formula is shown below.

[0071] As shown in Table 1, the MGF-MGO compound 1The H-NMR spectrum showed 9 proton signals, first, three new carbon signals (δC 67.82, δC 206.26 and δC 24.35), combined with the H14 to C-3, C-4, C-12 and H16 to C-14, C-15 related signals in the HMBC spectrum, based on these characteristics, finally determined the structure of MGF-MGO, a new kind of mangiferin derivatives, the structure is shown below.

[0072]

[0073] Example 2

[0074] Inhibition activity of mangiferin under simulated processing conditions on ACR, MGO and GO

[0075] (1) Experimental materials and instruments

[0076] Mangiferin (MGF, 95%, chromatographically pure, Shanghai Yuanye Biotechnology Co., Ltd.); Acrolein (ACR, 98% aqueous solution, analytical pure), methyl glyoxal (MGO) and glyoxal (GO) (40% aqueous solution, analytical pure, Shandong Shiyachemical Industry Co., Ltd.); Acetonitrile (chromatographically pure, Shanghai Reagent Company); Pure water (Hangzhou Wahaha Group Co., Ltd.); Sodium dihydrogen phosphate and sodium hydrogen phosphate are both analytical reagents (Shanghai Reagent Company); 2,4-dinitrophenylhydrazine hydrochloride (DNPH, 98%, Shanghai Tixiayicheng Industrial Development Co., Ltd.); o-phenylenediamine (DB, 98%, Shanghai Tixiayicheng Industrial Development Co., Ltd.)

[0077] High performance liquid chromatograph: Waters 2695 (Waters Corporation, USA); QL-861 vortex mixer (Ji Linbail Instrument Manufacturing Co., Ltd. in Jiangsu Haimen).

[0078] (2) Experimental steps

[0079] ACR, MGO, GO solution was prepared with 0.1 mol / L, pH 7.0 PBS, the concentration was 0.5 mmol / L, 0.422 g of morin standard was weighed, 18 mL of PBS solution was added, 2 mL of DMSO reagent was added to assist dissolution, and a 50 mmol / L stock solution was prepared, and then the morin stock solution was sequentially diluted to 0.5 / 1.0 / 1.5 mmol / L. In a 10 mL explosion-proof tube, 1 mL of ACR / MGO / GO solution (0.5 mmol / L) and 1 mL of morin solution (0.5 / 1.0 / 1.5 mmol / L / 3 mmol / L / 4 mmol / L / 5 mmol / L / 6 mmol / L) were added, and 1 mL of PBS solution was used instead of morin solution as a blank. After vortex mixing, the reaction was carried out at 100°C for 5, 10, 15 and 30 min in the dark. After the reaction, for ACR, 500 μL of the reaction solution was taken, 300 μL of DNPH solution was added, and the derivatization was carried out at 37°C for 2 h to calculate the ACR clearance rate; for MGO and GO, 500 μL of the reaction solution was taken, 100 μL of o-phenylenediamine solution was added, and the derivatization was carried out at 37°C for 2 h to calculate the MGO / GO clearance rate. The RCS content was detected by high performance liquid chromatography, and the inhibition rate of morin on RCS under simulated processing conditions was calculated. Each sample was made in triplicate.

[0080] The calculation formula of RCS clearance rate is:

[0081]

[0082] HPLC conditions:

[0083] Waters 2695 high performance liquid chromatograph (Waters Corporation, USA) was used for detection, Symmetry C18 column (250x4.6 mm, i.d. 5 μm) was selected, sample size was 10 μL, column temperature was 30°C. For ACR, the detection wavelength was 372 nm, the mobile phase A was acetonitrile, the mobile phase B was ultrapure water (containing 0.1% formic acid), the flow rate was 1.0 mL / min, and elution was carried out with 70% mobile phase A for 8.5 min. For MGO and GO, the detection wavelength was 315 nm, the mobile phase A was methanol, the mobile phase B was ultrapure water, the flow rate was 1.0 mL / min, and elution was carried out with 50% mobile phase A for 10 min.

[0084] (3) Experimental results

[0085] The reaction path mechanism diagram of morin with ACR, MGO, GO is shown in Figure 1 . The activity of morin in eliminating ACR, MGO and GO was studied by using a simulated food processing model (100°C, pH 7.0). First, the elimination of morin on the three kinds has a time and dose dependent relationship. The clearance rate of morin on ACR is fasterFigure 3 A). Compared with the blank group, the addition of 1.5 mmol / L mangiferin can remove more than 80% of ACR and 70% of MGO in 30 min Figure 3 A and Figure 3 B). When the amount of mangiferin added is increased, the removal rate of GO can also reach more than 80% in 30 min Figure 3 C). In summary, mangiferin can effectively remove ACR, MGO, and GO at the same time, and the removal effect is good, with broad spectrum.

[0086] Example 3

[0087] Simultaneous inhibition activity of mangiferin under simulated processing conditions on ACR, MGO, and GO

[0088] (1) Experimental materials and instruments

[0089] The same as Example 2.

[0090] (2) Experimental steps

[0091] A mixed solution of ACR, MGO, and GO was prepared with 0.1 mol / L PBS at pH 7.0, so that the final concentration of each was 0.5 mmol / L; 0.422 g of mangiferin standard was weighed and added to 18 mL of PBS solution, and 2 mL of DMSO reagent was added to assist dissolution, to prepare a 50 mmol / L stock solution; the mangiferin stock solution was then diluted to 0.5 / 1.0 / 1.5 mmol / L. In a 10 mL explosion-proof tube, 1 mL of the ACR-MGO-GO mixed solution (0.5 mmol / L each) and 1 mL of the mangiferin solution (0.5 / 1.0 / 1.5 mmol / L) were added, and 1 mL of PBS solution was added instead of the mangiferin solution as a blank. After vortex mixing, the reaction was carried out at 100°C in the dark for 5, 10, 15, and 30 min. After the reaction was completed, for ACR, 500 μL of the reaction solution was taken and 300 μL of DNPH solution was added for derivatization at 37°C for 2 h to calculate the ACR removal rate; for MGO and GO, 500 μL of the reaction solution was taken and 100 μL of o-phenylenediamine solution was added for derivatization at 37°C for 2 h to calculate the MGO / GO removal rate. The RCS content was detected by high performance liquid chromatography, and the inhibition rate of mangiferin under simulated processing conditions was calculated. Each sample was run in triplicate.

[0092] RCS removal rate calculation formula:

[0093]

[0094] HPLC conditions:

[0095] The same as Example 2.

[0096] (3) Experimental results

[0097] The activity of mangiferin in simultaneously clearing ACR, MGO, and GO was studied using a simulated food processing model (100°C, pH 7.0). First, the elimination of these three RCS by mangiferin was time- and dose-dependent. Unlike Example (2) in which RCS were cleared separately, when ACR, MGO, and GO were present in the reaction system at the same time, mangiferin had the highest clearance rate for MGO ( Figure 4 ). Compared with the blank group, when the addition amount of mangiferin is 0.5mmol / L, the clearance rate of MGO can reach 40%; adding 1.5mmol / L mangiferin can clear up to 75% of MGO, which exceeds the effect of clearing MGO alone under the same conditions (70%), and when the three are inhibited simultaneously, a low concentration of mangiferin (1.5mmol / L) can achieve the effect of high concentration mangiferin inhibiting GO (5mmol / L), which can reach 60% clearance rate. In fact, when the three are inhibited together, the total amount of mangiferin is 1.5mmol / L, and the amount of mangiferin that acts on each RCS will be even less, and the average distribution is only 0.5mmol / L, which effectively reflects that low-dose mangiferin can have a significant effect when the three are inhibited together; at the same time, the clearance rate of ACR can still be maintained at a high level of more than 75%. The emergence of the above situation may be due to the emergence of new products for further inhibition. The experiment of this example shows that the present invention can use a low concentration of mangiferin to inhibit three RCSs simultaneously, which can not only ensure the ACR inhibition effect, but also better inhibit the difficult-to-treat MGO and GO, and the effect is significant.

[0098] Example 4

[0099] Synchronous inhibition mechanism of ACR, MGO and GO under simulated processing conditions by mangiferin

[0100] (1) Experimental materials and instruments

[0101] Same as Example 2.

[0102] (2) Experimental steps

[0103] ACR, MGO, and GO solutions were prepared in 0.1 mol / L, pH 7.0 PBS at concentrations of 50, 150, and 500 mmol / L, respectively. 0.422 g of mangiferin standard was weighed and added to 18 mL of PBS. 2 mL of DMSO was then added for solubilization to a 50 mmol / L solution. Three 10 mL explosion-proof tubes were used to add 1 mL of ACR-MGO-GO solution (50 mmol / L, 150 mmol / L, and 500 mmol / L) and 1 mL of mangiferin solution (50 mmol / L), respectively, to achieve reaction solution concentration ratios of 1:1, 1:3, and 1:10. 1 mL of PBS was used instead of the RCS solution as a blank. After vortex mixing, the reaction was incubated at 100°C in the dark for 5, 10, 15, and 30 min. After completion of the reaction, the reaction was immediately terminated in an ice bath. A 100 μL sample was diluted to 200 ng / mL and analyzed by UPLC-MS / MS (same as in Example 1).

[0104] (3) Experimental results

[0105] At 100°C, Figure 5 , Figure 6 As shown in Figure 2, when MGF:ACR:MGO:GO=1:1:1:1 and the reaction time is 5 minutes, three new peaks (Rt=9 min, Rt=11 min, Rt=12.8 min) appeared compared with Example 2. The two peaks at Rt=9 min and Rt=12.8 min have similar fragments m / z 549.1242 [MH] - , than MGF-ACR m / z 477.1033[MH] - 72 more amu (i.e., MW MGO,72), than MGF m / z 421.0771 [MH] - More than 128amu (ie MW MGO+MW ACR, 72+56) In addition, they are in MS 2 The secondary mass spectrometry produced m / z 477.1033 [MH] - ,m / z 301[MH] - The two main molecular fragments were produced by the loss of one MGO molecule (m / z 72) and one C4H8O4 molecule, respectively. These results indicate that the two new products are isomers of the adduct formed by MGF-ACR capturing another MGO, namely MGF-ACR-MGO-1 and MGF-ACR-MGO-2; for the peak at Rt = 11 min, m / z 607.1299 [MH] - , than MGF-2MGO m / z 565.1193[MH] -one more 58 amu (MW GO, 58) and one water molecule (MW H2O, 18) less In addition, they produced m / z 565.1193 [M-H] in MS 2 m / z 565.1193 [M-H] - m / z 343.0472 [M-H] - These two fragments were produced by the loss of one molecule of GO (m / z 58), and one molecule of C5H 10 For the three new substances produced, it can be better explained that the inhibitory activity of Example 3, compared with the inhibitory activity of MGF on ACR alone, when used in combination, MGF-ACR has captured one molecule of MGO, not only has higher inhibitory activity on ACR, but also enhances the inhibitory effect on MGO, but the inhibitory effect on ACR remains at a high level; in addition, when used in combination, MGF and MGO form a di-adduct, while continuing to capture one molecule of GO, which also better explains the reason for the increase in the inhibition rate of MGO and GO when used in combination.

[0106] Example 5

[0107] Inhibitory activity of MGF-ACR on ACR, MGO and GO under simulated processing conditions

[0108] (1) Experimental materials and instruments

[0109] The same as Example 2

[0110] (2) Experimental steps

[0111] ACR, MGO and GO solutions were prepared with 0.1 mol / L, pH 7.0 PBS, all at a concentration of 0.5 mmol / L, 0.002 g of MGF-ACR sample (prepared in Example 1) was weighed, 2.5 mL of PBS solution was added, and 0.5 mL of DMSO reagent was added to assist dissolution, to prepare a 1.5 mmol / L solution, 1 mL of ACR / MGO / GO solution (0.5 mmol / L) and 1 mL of MGF-ACR solution (1.5 mmol / L) were added in a 10 mL explosion-proof tube, and 1 mL of PBS solution was used instead of MGF-ACR solution as a blank. After vortex mixing, react at 100℃ for 30 min in the dark. After the reaction was completed, for ACR, 500 μL of the reaction solution was taken, 300 μL of DNPH solution was added, and the derivative was calculated at 37℃ for 2h to calculate the ACR clearance rate; for MGO and GO, 500 μL of the reaction solution was taken, 100 μL of o-phenylenediamine solution was added, and the derivative was calculated at 37℃ for 2h to calculate the MGO / GO clearance rate. The ACR / MGO / GO content was detected by high performance liquid chromatography, and the inhibition rate of MGF-ACR on ACR / MGO / GO under simulated processing conditions was calculated. Each sample was done in triplicate.

[0112] The formula for calculating the RCS clearance rate is:

[0113]

[0114] HPLC conditions:

[0115] The same as Example 2

[0116] (3) Experimental results

[0117] The simulated food processing model (100℃, pH 7.0) was used to study the clearance activity of MGF-ACR on ACR, MGO and GO. According to Figure 7 It can be found that there is no significant difference in the inhibition effect of ACR between the addition of equal concentration of MGF and MGF-ACR, and the clearance efficiency is still high. When equal concentration of MGF and MGF-ACR is added, the inhibition effect of MGF-ACR on MGO is obviously higher than that of MGF, and there is a significant difference. Secondly, it is proved in Example 2 that the addition amount of MGF needs to reach 6 mmol / L in order to achieve an inhibition effect of about 80% on GO, while MGF-ACR only needs to add 1.5 mmol / L to achieve the same effect, and is significantly better than the inhibition effect of 1.5 mmol / L of MGF on GO. Therefore, the adduct of MGF and ACR in the present application has excellent inhibition activity on carbonyl compounds, and is significantly better than MGF.

[0118] Example 6

[0119] Study on the scavenging effect of mango peel on active carbonyl compounds generated during the baking process of heavy oil cakes

[0120] (1) Experimental materials and instruments

[0121] Green mango peel, mangiferin (MGF, 95%, chromatographically pure, Shanghai Yuanye Biotechnology Co., Ltd.); low-gluten flour (Shanghai Yihai Kerry International Trade Co., Ltd.); salt, white sugar, butter, baking powder and eggs (Hongli A Jia Supermarket in Suqian City, Jiangsu Province).

[0122] Waters 2695 high performance liquid chromatograph (Waters Corporation, USA); ZQTY-70 table type oscillation incubator (Shanghai Zhichu Instrument Co., Ltd.); QL-861 vortex mixer (Ji Linbel Instrument Manufacturing Co., Ltd. in Haian City, Jiangsu Province); KQ-300B ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); water bath (Jintan Digital Constant Temperature Water Bath); CRTF32K oven (Changdi Electrical Appliance Co., Ltd.).

[0123] (2) Experimental steps

[0124] ①Preparation of heavy oil cake

[0125] According to the literature "Valorization of Peel-Based Agro-Waste Flour for Food Products: A Systematic Review on Proximate Composition and Functional Properties", mango peel is added to bread, biscuits and other foods. In this example, the effect of mangiferin is demonstrated. First, 30.0 g of butter is slightly softened by heating, and 4 / 8 / 10% (calculated by the mass ratio of low-gluten flour) of mango peel or 7.2 mg, 14.4 mg, and 18 mg of pure mangiferin is added, respectively. Then, 30.0 g of white granulated sugar is added, and the two are stirred to a slight whiteness with a handheld electric mixer. 30.0 g of fresh whole egg liquid is added in portions, and after stirring evenly, 30.0 g of low-gluten flour, 0.3 g of salt, and 0.6 g of baking powder are added. Then, a handheld electric mixer is used to stir for 4 min to obtain the cake batter needed for making heavy oil cake. The cake batter is transferred to a mold on a baking tray and placed in a preheated oven. The temperature of the upper and lower heat is 175°C, and the cake is baked for 45 min. After cooling, the heavy oil cake is crushed and stored in a -80°C freezer for later use. All samples are prepared in triplicate.

[0126] ②Study on the RCS scavenging activity of mangiferin in heavy oil cake system

[0127] Take 0.5 g of heavy oil cake crushed sample in a centrifuge tube, add 5 mL of a mixture of methanol and PBS (pH 7.0, 0.1 mol / L) solution 1:1 (50:50, v / v), and vortex mix for 3 min, then centrifuge at 8000 r / min for 10 min. After centrifugation, 500 μL of sample solution is added to 300 μL of DNPH solution, vortex mixed, and then derivatized at 37°C for 2 h to calculate the ACR scavenging rate; 1 mL of sample solution is added to 200 μL of o-phenylenediamine solution, vortex mixed, and then derivatized at 37°C for 2 h to calculate the MGO and GO scavenging rates. The detection method is the same as in Example 2, and all samples are prepared in sextuplicate.

[0128] (3) Experimental results

[0129] As shown in Figure 8 A, the addition of different doses of mango peel to heavy oil cake shows a dose-dependent relationship in the scavenging activity of ACR, MGO, and GO. When 10% of mango peel is added to heavy oil cake, the inhibition rate of mango peel on MGO and GO reaches about 60%, and the inhibition rate on ACR reaches about 50%, as shown in Figure 8B; when adding pure mangiferin, the inhibition rate of ACR, MGO and GO in heavy oil cake was about 15% higher than that of adding peel, indicating that mangiferin has high efficiency in removing RCS when applied to actual food processing system.

Claims

1. Application of mangiferin in inhibiting reactive carbonyl compounds, including acrolein, methylglyoxal and glyoxal.

2. The use according to claim 1, characterized in that The mangiferin is used in the preparation of an inhibitor for inhibiting active carbonyl compounds produced in the environment and in food processing.

3. The use according to claim 1, characterized in that The application of mangiferin in the preparation of inhibitors for inhibiting active carbonyl compounds in organisms.

4. The use according to claim 1, characterized in that The mangiferin can inhibit various harmful addition or cross-linking products formed by the reaction of active carbonyl compounds with nucleophilic biological macromolecules in the preparation environment, food processing or application of acrolein, methylglyoxal, and glyoxal inhibitors in organisms.

5. The use according to claim 1, characterized in that The mangiferin can capture active carbonyl compounds to form addition products, thereby reducing the content of active carbonyl compounds.

6. The use according to claim 5, characterized in that The adduct product includes the monoadduct product of mangiferin and acrolein, MGF-ACR, whose structure is shown below: ； The adduct includes the monoadduct of mangiferin and methylglyoxal, MGF-MGO, the structure of which is shown below: 。 7. The use according to claim 1, characterized in that The mangiferin is used as the sole ingredient, or as the main ingredient in combination with other substances to form a preparation for the preparation of a broad-spectrum inhibitor of carbonyl compounds.

8. Use of a monoadduct of mangiferin and a reactive carbonyl compound in inhibiting reactive carbonyl compounds, wherein the reactive carbonyl compound includes acrolein, methylglyoxal, and glyoxal. The monoadduct is the monoadduct of mangiferin and acrolein, MGF-ACR, and its structure is shown below: ； Alternatively, the monoadduct is the monoadduct of mangiferin and methylglyoxal, MGF-MGO, the structure of which is shown below: 。 9. An inhibitor of active carbonyl compounds in the environment, food processing or in an organism, characterized in that: The inhibitor uses mangiferin and / or the monoadduct according to claim 8 as the sole component, or as the main component, and is used in combination with other substances to form a preparation. The active carbonyl compound includes acrolein, methylglyoxal and glyoxal.

10. Use of mango peel and / or mango leaf in the preparation of an inhibitor for inhibiting reactive carbonyl compounds, wherein the main component of the mango peel and / or mango leaf is mangiferin, and the reactive carbonyl compounds include acrolein, methylglyoxal and glyoxal.

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