CN104592331A - Quercetin derivative as well as preparation process and application thereof - Google Patents

Abstract

The invention belongs to the field of analysis and application of tree secondary metabolism active ingredients and particularly relates to a novel quercetin derivative, namely, quercetin-3-O-(4''-galloyl)-alpha-L-rhamnopyranoside, which is separated from sophorajaponica of leguminosae and an application of the quercetin derivative in preparation of anti-oxidation products. The liquid-liquid extraction and modern chromatographic separation preparation process are simple and normative and is easy to operate and industrially produce.

Description

A kind of quercetin derivative and preparation technology thereof and application

Technical field

The invention belongs to trees secondary metabolism active fraction preparation to resolve and Application Areas, be specifically related to be separated from pulse family (Leguminosae) arbor Chinese scholar tree (Sophora japonica Linn.) obtain a kind of new Quercetin (Quercetin) derivative and Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides (Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranoside) and to apply.

Background technology

Chinese scholar tree (Sophora japonica Linn.), having another name called middle Chinese scholar tree, Chinese scholartree, fructus sophorae etc., is China's endemic tree that pulse family (Leguminosae) Sophora (Sophora Linn.) integrates medicinal, edible, material is used and viewed and admired.Very wide in the cultivated area of China, northern from THE SOUTH OF NORTHEAST CHINA, northwest to Shaanxi, SOUTH OF GANSU, southwest to Sichuan, Yunnan below height above sea level 2600m, reach in the south the ground such as Guangdong, Guangxi (Sun Rongxi, etc., Jouranl of Agricultural University of Hebei, 2011,34 (3): 65-70).

The present invention utilizes liquid-liquid extraction and chromatographic separation technology from Chinese scholar tree (Sophora japonica Linn.), prepare a kind of new quercetin derivative and Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides, and this quercetin derivative English Quercetin-3-O-by name (4 "-galloyl)-α-L-rhamnopyranoside.

Summary of the invention

The object of this invention is to provide a kind of new quercetin derivative and Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides (Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranoside).

Another object of the present invention is to provide the technique preparing Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides from pulse family (Leguminosae) Sophora (Sophora Linn.) arbor Chinese scholar tree (Sophorajaponica Linn.).

3rd object of the present invention is to provide anti-oxidant activity effect application of Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides.

Technical scheme of the present invention is summarized as follows:

The quercetin derivative of following chemical structural formula:

Compound molecule formula of the present invention is C ₂₈h ₂₄o ₁₅, chemistry Quercetin-3-O-by name (4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides, the English of its correspondence is called Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranoside.

From Chinese scholar tree (Sophora japonica Linn.), prepare the technique of Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides, comprise the steps:

(1) the Chinese scholar tree root skin plant material of pulverizing is got, be 1: 1 ~ 1: 10 acetonitrile solutions adding that concentration expressed in percentage by volume is 40% ~ 60% in mass ratio, heating or normal temperature or microwave or ultrasonic-assisted extraction 1 ~ 5 time, each 1 ~ 48 hour, filter, filtrate reduced in volume, to 1% ~ 15% of original volume, obtains crude extract;

(2) add the water of crude extract quality 1 ~ 5 times, stir, add the n-hexane extraction 1 ~ 5 time of crude extract quality 1 ~ 5 times, be separated, residue water layer is added the chloroform extraction 1 ~ 5 time of crude extract quality 1 ~ 5 times, be separated, chloroform layer concentrating under reduced pressure is obtained chloroform extraction phase;

(3) chloroform extraction prepares Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides through at least one of gel filtration chromatography and silica gel column chromatography.

This preparation technology has working method simple specification, yield is high, preparation cost is low feature.

Experiment confirms that Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides has good Scavenging activity (IC to DPPH free radical ₅₀value is 6.56 μMs), its elimination effect is better than control group α-Tocopherol (IC ₅₀value is 6.82 μMs) and BHT (IC ₅₀value is 6.88 μMs).This shows that the anti-oxidant activity of Quercetin-3-O-(the 4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides prepared is remarkable, can be used as antioxidant and is applied to food, medicine, healthcare products and cosmetic industry.

Embodiment

With reference to the following example by easier, comprehend the present invention, providing embodiment is to illustrate the present invention, instead of limits the present invention by any way.

Embodiment 1:

The preparation of Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides

(1) getting the Chinese scholar tree root skin plant material of pulverizing, is 1: 4 acetonitrile solution adding that concentration expressed in percentage by volume is 45% in mass ratio, extract at room temperature 4 times, each 36 hours, and filter, filtrate reduced in volume, to 5% of original volume, obtains crude extract;

(2) add the water of crude extract quality 2 times, stir, add the n-hexane extraction 2 times of crude extract quality 2 times, be separated, residue water layer is added the chloroform extraction 2 times of crude extract quality 2 times, be separated, chloroform layer concentrating under reduced pressure is obtained chloroform extraction phase;

(3) chloroform extraction prepares Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides through silica gel column chromatography.

Embodiment 2:

The preparation of Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides

(1) getting the Chinese scholar tree root skin plant material of pulverizing, is 1: 3 acetonitrile solution adding that concentration expressed in percentage by volume is 55% in mass ratio, microwave extraction 3 times, each 2 hours, and filter, filtrate reduced in volume, to 6% of original volume, obtains crude extract;

(2) add the water of crude extract quality 3 times, stir, add the n-hexane extraction 3 times of crude extract quality 3 times, be separated, residue water layer is added the chloroform extraction 3 times of crude extract quality 3 times, be separated, chloroform layer concentrating under reduced pressure is obtained chloroform extraction phase;

(3) chloroform extraction prepares Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides through gel filtration chromatography.

The Structural Identification of this compound:

This compound ¹proton signal [δ in H NMR spectrogram (data are as table 1) _h6.23 (1H, d, J=2.3Hz, H-6) and δ _h6.41 (1H, d, J=2.3Hz, H-8)] show that the A ring of this compound is Phloroglucinol type aromatic nucleus.And its ¹³c NMR signal [δ _c159.6 (C-2), 135.3 (C-3), 179.5 (C-4)] to disclose this compound be typical flavonol.Proton peak [the δ of this compound _h7.65 (1H, d, J=2.2Hz, H-2 '), δ _h6.92 (1H, d, J=8.3Hz, H-5 ') and δ _h7.52 (1H, dd, J=8.3Hz, J=2.2Hz)] the B ring that discloses this compound is typical ABX formula aromatic nucleus.Therefore, the aglycon of this compound is confirmed as Quercetin.

At it ¹during H NMR composes, 1 terminal hydrogen δ _h5.31 (1H, br s, H-1 "), methyl δ _h1.02 (3H, d, J=6.3Hz, H-6 ") and four proton δ _h3.53-5.08 (4H, H-2 ", 3 ", 4 " and, 5 ") show in this compound molecule containing a α-L-rhamnopyranosyloxyhy glucosides.

This compound ¹³c NMR signal [δ _c121.8 (C-1 " '), 109.9 (C-2 " ', 6 " '), 146.3 (C-3 " ', 5 " '), 139.8 (C-4 " ')] in conjunction with proton peak δ _h7.10 (2H, s, H-2 " ', 6 " ') show in this compound molecule, to comprise a galloyl.

The rhamnosyl terminal hydrogen δ of this compound of HMBC spectrum display _h5.31 (the carbon signal δ of 1H, br s, H-1 ") and quercetin aglycon _c135.3 (C-3) have Long-Range Correlation, determine that α-L-rhamnopyranosyloxyhy glucosides is connected with quercetin aglycon C-3.And the proton signal δ of rhamnosyl _h5.08 (1H, dd, J=10.1Hz, J=9.8Hz, H-4 ") and the carbon peak δ on galloyl _c168.4 (C-7 " ') there is Long-Range Correlation, then determine α-L-rhamnopyranosyloxyhy glucosides 4 " position and 7 of galloyl " ' position and be connected.

As shown in table 1, the undistorted polarization transfer of this compound strengthens collection of illustrative plates (DEPT spectrum) and shows altogether containing 28 carbon in its molecule, wherein, and 1, methyl carbon, mesomethylene carbon 0, methine carbon 12, quaternary carbon 15.The PositiveFAB mass spectrum of this compound shows its m/z [M+H] ⁺and m/z [M+Na] ⁺respectively corresponding 601 and 623, more comprehensive its ¹h and ¹³c NMR spectrum elucidation must the molecular formula of this compound be C ₂₈h ₂₄o ₁₅.

To sum up, the structure of this compound is confirmed as Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides.

Through retrieval, this compound is the new quercetin derivative never reported before this.This compound is Yellow amorphous powder, and fusing point is 169 ~ 171 DEG C, specific rotation this compound has uv-absorbing under ultraviolet lamp 254nm wavelength, and the color reaction of spraying of the iron trichloride ethanolic soln (mass percent) with 1% is deep green.TLC launches in test, with the trimethyl carbinol-glacial acetic acid-water volume ratio 3: 1: 1 for solvent systems, and the R of this compound _fvalue is 0.66, and when launching with the solvent systems of glacial acetic acid-water volume ratio 6: 94, the R of this compound _fvalue is 0.25.

Table 1. Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides ¹h (400MHz), ¹³c NMR (100MHz) and DEPT modal data (MeOD)

Embodiment 3:

The anti-oxidant activity evaluation of Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides

1. product to be evaluated: the Quercetin-3-O-of above-mentioned preparation (4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides purity after HPLC measures is 99.9%, selects α-Tocopherol and BHT to be positive control group.

2. experimental technique: the anti-oxidant activity merit rating of sample selects the removing DPPH free radical test method of widespread use (DPPH free radical scavenging method), concrete operations strictly carry out (Si CL to describe step in document, et al, PlantaMedica, 2009,75,1165-1167).Each group of experiment all independent parallelly carries out three times, gets its mean value calculation.

Experimental result is as shown in table 2.Result shows, Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides is to the Scavenging activity IC of DPPH free radical ₅₀value is 6.56 μMs, and positive control group α-Tocopherol and BHT free radical scavenging IC ₅₀value is respectively 6.82 and 6.88 μMs.Testing data discloses Quercetin-3-O-(4 "-galloyl) Scavenging activity of-α-L-rhamnopyranosyloxyhy glucosides to DPPH free radical and is better than positive control group α-Tocopherol and BHT.The anti-oxidant activity of Quercetin-3-O-(the 4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides of preparation is remarkable, can be used as antioxidant and is applied to food, medicine, healthcare products and cosmetic industry.

Table 2. Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides, BHT and α-Tocopherol scavenging ability of DPPH free radical (IC ₅₀value, μM)

Claims (3)

1. the compound of following structural formula:

Chemistry Quercetin-3-O-by name (4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides, the English of its correspondence is called Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranoside.

2. the preparation method of claim 1 compound, is characterized in that comprising the steps:

(1) the Chinese scholar tree root skin plant material of pulverizing is got, be 1: 1 ~ 1: 10 acetonitrile solutions adding that concentration expressed in percentage by volume is 40% ~ 60% in mass ratio, normal temperature or heating or microwave or ultrasonic extraction 1 ~ 5 time, each 1 ~ 48 hour, filter, filtrate reduced in volume, to 1% ~ 15% of original volume, obtains crude extract;

(2) add the water of crude extract quality 1 ~ 5 times, stir, add the n-hexane extraction 1 ~ 5 time of crude extract quality 1 ~ 5 times, be separated, residue water layer is added the chloroform extraction 1 ~ 5 time of crude extract quality 1 ~ 5 times, be separated, chloroform layer concentrating under reduced pressure is obtained chloroform extraction phase;

(3) chloroform extraction prepares Quercetin-3-O-(4 "-galloyl)-α-L-rhamnopyranosyloxyhy glucosides through at least one of gel filtration chromatography and silica gel column chromatography.

3. claim 1 compound is preparing the application in oxidation resistant product.