CN102731521A

CN102731521A - Chromone compound in aromatic tobaccos, preparation method and application thereof

Info

Publication number: CN102731521A
Application number: CN2012102333170A
Authority: CN
Inventors: 杨光宇; 段沅杏; 赵伟; 王岚; 陈永宽; 缪明明
Original assignee: Yunnan Academy of Tobacco Science
Current assignee: Yunnan Academy of Tobacco Science
Priority date: 2012-07-06
Filing date: 2012-07-06
Publication date: 2012-10-17

Abstract

The invention relates to a chromone compound in aromatic tobaccos, a preparation method of the chromone compound and application of the chromone compound, which belong to the field of tobacco chemistry, in particular to a novel chromone compound contained in the aromatic tobaccos, a preparation method of the novel chromone compound and an antibacterial activity of the novel chromone compound. The preparation method comprises the following steps of: pulverizing an aromatic tobacco sample, and then carrying out ultrasonic extraction by using 95% alcohol for three times; merging the extraction solutions, decompressing and concentrating the obtained merged extraction solution into an extract; carrying out primary chromatography and fractionation on the extract by using a silicagel column; and then carrying out further separation by adopting a high-pressure liquid chromatogram method to obtain the required compound, namely tobchromone A. The compound has a simple structure, can be artificially synthesized easily, has good antibacterial activity and exerts antibacterial effects on ordinary proteus, escherichia coli, staphylococcus, bacillus subtilis, and the like.

Description

Chromone compounds in a kind of Turkish tobaccos

Technical field

The invention belongs to the tobacco chemistry field, more particularly, the present invention relates to contained chromone compounds and preparation method thereof in a kind of new Turkish tobaccos; Simultaneously, the anti-microbial activity that also relates to this compound.

Background technology

Tobacco is to contain maximum a kind of of chemical substance in human each kind of plant of being familiar with, and through the research of decades, people identify from tobacco that at present the monomer chemical substance of coming out just surpasses kind more than 3000, and also have many compositions to identify as yet.Tobacco also can therefrom be extracted the multiple chemical ingredients that the value utilized is arranged except that being mainly used in the cigarette smoking purposes, therefrom find to have the guiding compound of value of exploiting and utilizing.Therefore, except that consuming as cigarette, the research of strengthening other purposes of tobacco is also significant.

Turkish tobaccos are claimed Turkey's cigarette, east type cigarette again, originate in Mediterranean Sea coastwise contries, are a kind of special tobacco types of safflower tobacco (Nicotiana tobacum).Because Turkish tobaccos have strong fragrance and pure jealous quality characteristic, are one of important source material of production mixed type, outer odor type and oriental type cigarette and pipe tobacco.In China, Turkish tobaccos have establishing in large scale in the Baoshan, Yunnan at present.

Chromone (chromone) is the biologically active substance that one type of occurring in nature extensively exists, because of the most coloured chromone of gaining the name of this compounds.Because plant chromone constituent structure type is many, stereochemistry is complicated, has multiple biological activity, and is very active to the research in this field both at home and abroad; No matter be naturally occurring, or the chromone compounds that obtains of synthetic, chemist's extensive concern all caused.But through the present report that does not also have chromone compounds in the Turkish tobaccos of document retrieval.

Summary of the invention

The object of the present invention is to provide the chromone compounds that separation obtains in the Turkish tobaccos of the Baoshan, a kind of Yunnan.

Another object of the present invention provides a kind of preparation method of said compound.

Further purpose of the present invention provides the anti-microbial activity of said compound.

The object of the invention is achieved through following technical proposals.

A. the present invention separates from the Turkish tobaccos of the Baoshan, Yunnan and obtains chromone compounds, and this compound has following structural formula:

，

The called after of this compound: tobacco chromone A (Tobchromone A).

B. the invention provides the preparation method of described chromonic compound, this method adopts following step to accomplish:

(1) the Turkish tobaccos sample is pulverized the back and is divided with the ethanol of 95 % V/V and use supersound extraction 3～5 times, united extraction liquid, and concentrating under reduced pressure becomes medicinal extract;

(2) medicinal extract just divides with silica gel column chromatography, adopts HPLC further to separate then, promptly obtains required compound.

The concrete steps of the inventive method are:

(1) is crushed to 20～40 orders with Turkish tobaccos;

(2) be solvent with 95%V/V ethanol, with supersound extraction 3～5, each 30～60 minutes;

(3) medicinal extract is with 80～100 purpose silica gel mixed samples; Carry out the silica gel column chromatography rough segmentation with 80～200 purpose silica gel dry column-packings; With the quality proportioning that chloroform-acetone is pressed 20:1,9:1,8:2,7:3,5:5 respectively, carry out gradient elution, collect various piece elutriant and concentrated; The 8:2 part of elutriant is further used HPLC (special-purpose in the industry) separation and purification, adopts the C of 20 mm * 250 mm ₁₈Chromatographic column, flow velocity is 15 mL/min, is moving phase with the methanol, progressively adjusts the two ratio and segments step by step, promptly obtains required chromone compounds.

C. this compound has been carried out the anti-microbial activity screening, it has significant activity to proteus vulgaris, intestinal bacteria, staphylococcus, subtilis, sarcina, penicillium spp, Aspergillus flavus and black-koji mould etc.; Wherein the bacteriostasis rate to proteus vulgaris, intestinal bacteria, staphylococcus and subtilis reaches more than 85%.

The invention has the beneficial effects as follows: The compounds of this invention is simple in structure, and synthetic realizes that easily the compound anti-microbial activity is good, and the bacteriostasis rates of proteus vulgaris, intestinal bacteria, staphylococcus, subtilis etc. is reached more than 85%; Can be used as the guiding compound of anti-microbial type drug development.

Description of drawings

Fig. 1 be The compounds of this invention proton nmr spectra ( ¹H NMR) figure.

Fig. 2 be The compounds of this invention carbon-13 nmr spectra ( ¹³C NMR) figure.

Fig. 3 is the main HMBC correlogram of The compounds of this invention.

Embodiment

In order to make the object of the invention, technical scheme and advantage clearer,, the present invention is further elaborated below in conjunction with accompanying drawing and embodiment.Should be appreciated that specific embodiment described herein only in order to explanation the present invention, and be not used in qualification the present invention.

Embodiment 1---and compound extracts and separates

The Turkish tobaccos sample is adopted in the Baoshan, Yunnan, and kind is the Bath horse.Turkish tobaccos complete stool sampling 1.5～2.5 kg are crushed to 20～40 orders, and with supersound extraction 3 times, each 30～60 minutes, extracting solution merged, filtration with the ethanol of 95%V/V, and concentrating under reduced pressure becomes medicinal extract.Medicinal extract is mixed appearance with an amount of methyl alcohol (medicinal extract weight 1.5 times) dissolving back with the thick silica gel of 80～120 g (80～100 order), and 1.5～2.0 kg silica gel (80～200 order) are adorned post and carried out silica gel column chromatography.Chloroform-acetone (gradient elution of 20:1 → 1:1), the TLC monitoring merges identical part, obtains 5 parts; Wherein chloroform-acetone (8:2) wash-out partly separates with the prompt logical sequence 1,100 half preparation HPLies of peace; Methyl alcohol with 46% is moving phase, the C of (20 mm * 250 mm, 5 μ m) ₁₈Preparative column is a stationary phase, and flow velocity is 15 mL/min, and it is 254 nm that UV-detector detects wavelength, and each sample introduction 200 μ L collect the chromatographic peak of 22.8 min, and the back evaporate to dryness repeatedly adds up; The gained material is used pure dissolve with methanol once more, is moving phase with pure methyl alcohol again, separates with Sephadex LH-20 gel filtration chromatography, get final product this new compound.

Embodiment 2---the evaluation of compound:

Table-1 compound ¹H with ¹³C NMR data

position	C?(mult.)?	H?(mult, J,?Hz)
			2	167.2?s
3	108.0?d	6.34,?s
			4	180.9?s
5	161.3?s
			6	100.5?d	6.21,?s
7	159.2?s
			8	101.8?s
9	158.4?s
			10	107.4?s
11	36.2?t	2.57,?d,?J?=?8.1
			12	32.0?t	1.89,?m
13	62.7?t	3.58,?d,?J?=?6.3
			14	115.1?d	6.63,?d,?J?=?10.0
15	127.8?d	5.64,?d,?J?=?10.0
			16	77.8?s
17,?18	27.7?q	1.50,?s
			Ar-OH		10.73?brs

This patent compound is orange-yellow jelly; UV spectrum (solvent is a methyl alcohol), λ _Max(log ε) 222 (4.51), 245 (3.97), 268 (4.22), 358 (4.02) nm; Ir spectra (pressing potassium bromide troche) v _Max3392,2962,2881,1726,1662,1623,1584,1426,1375,1329,1284,1172,1128,853 cm ^-1High resolution mass spectrum (HRESIMS) provides quasi-molecular ion peak m/z 301.1067 [M-H] ^-(calculated value 301.1076).In conjunction with ¹H with ¹³C NMR spectrum provides a molecular formula C ₁₇H ₁₈O ₅, degree of unsaturation is 9.From ¹H with ¹³CNMR spectrum (figure-1 and figure-2, attribution data is seen table-1) signal can find out in the compound except that the chromone parent, 1 phenolic hydroxyl group signal in addition, 1 group of 3-hydroxypropyl signal, 1 group of gem-dimethyl chromene ring signal.Can infer that from HMBC is relevant phenolic hydroxyl group is substituted in the C-5 position of chromone ring, the 3-hydroxypropyl is substituted in the C-2 position of chromone ring, and gem-dimethyl chromene ring is substituted in the C-7 and the C-8 position of chromone ring, and the structure of compound obtains confirming since then.This compound is named as tobacco chromone A.

Embodiment 3---and anti-microbial activity detects and uses:

The in-vitro antibacterial experiment is carried out with agar diffusion method, and test strain is proteus vulgaris, intestinal bacteria, staphylococcus, subtilis, sarcina, penicillium spp, Aspergillus flavus, black-koji mould.At first will be tried on the flat board that bacterium is coated in plain agar substratum (Carnis Bovis seu Bubali cream, peptone, sodium-chlor, serum, agar) equably; (The compounds of this invention dissolves with 10 mL DMSO with testing compound again; Thin up becomes the solution of 50 g/mL) soaked tablet (diameter 5 mm) is placed on the germ-carrying substratum; Put into thermostat container, observe the inhibition zone size after hatching 24-72 h in 25 ℃.The result shows: The compounds of this invention has very strong activity to proteus vulgaris, intestinal bacteria, staphylococcus and subtilis, and inhibiting rate surpasses 85%, and other experimental strain is also had tangible resistance.

The compounds of this invention has been carried out safety evaluation,, proved that The compounds of this invention is nontoxic to animal through mouse bone marrow cells micronucleus test, Ames experiment and TK transgenation experiment, safe in utilization.

This compound is added on the cigarette tipping paper with the concentration of 50 g/mL; By the detection method of the GB15979-2002 of the People's Republic of China's " disposable use sanitary product hygienic standard ", get the cigarette that added The compounds of this invention and use tipping paper, 2.0 * 3.0 mm size, bacterial detection sum, total number of fungi.The result shows that the tipping paper total number of bacterial colony of adding The compounds of this invention obviously reduces, and the bacteriums of several kinds of tests are all had obvious restraining effect, and the bacteriostasis rate of proteus vulgaris, intestinal bacteria, staphylococcus and subtilis is all reached more than 72%.

Claims

1. the middle chromone compounds that grows tobacco is characterized in that: this compound separates from the Turkish tobaccos of the Baoshan, Yunnan and obtains, and structural formula is:

?，

The called after of this compound, tobacco chromone A (Tobchromone A).

2. the preparation method of chromone compounds in the said tobacco of claim 1 is characterized in that, this method adopts following steps preparation:

3. by the preparation method of chromone compounds in the said tobacco of claim 2, it is characterized in that concrete steps are:

(1) is crushed to 20～40 orders with Turkish tobaccos;

(3) medicinal extract is with 80～100 purpose silica gel mixed samples; Carry out the silica gel column chromatography rough segmentation with 80～200 purpose silica gel dry column-packings; With the quality proportioning that chloroform-acetone is pressed 20:1,9:1,8:2,7:3,5:5 respectively, carry out gradient elution, collect various piece elutriant and concentrated; The 8:2 part of elutriant is further used the HPLC separation and purification, adopts the C of 20 mm * 250 mm ₁₈Chromatographic column, flow velocity is 15 mL/min, is moving phase with the methanol, progressively adjusts the two ratio and segments step by step, promptly obtains required chromone compounds.

In the described tobacco of claim 1 chromone compounds to the application in proteus vulgaris, intestinal bacteria, staphylococcus, subtilis, sarcina, penicillium spp, Aspergillus flavus and black-koji mould antibiotic.