CN102323378A - Method for rapidly detecting proanthocyanidin in plant - Google Patents
Method for rapidly detecting proanthocyanidin in plant Download PDFInfo
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- CN102323378A CN102323378A CN201110250289A CN201110250289A CN102323378A CN 102323378 A CN102323378 A CN 102323378A CN 201110250289 A CN201110250289 A CN 201110250289A CN 201110250289 A CN201110250289 A CN 201110250289A CN 102323378 A CN102323378 A CN 102323378A
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Abstract
The present invention relates to a method for rapidly detecting proanthocyanidin in a plant organ or a plant tissue. According to the method, with adopting a dyeing method, a dye is added to carry out dyeing for the plant organ or the plant tissue, wherein the dye can be specifically combined with the proanthocyanidin; with an acid-alcohol solution of DMACA (p-dimethylaminocinnamaldehyde), the color of the proanthocyanidin-containing plant organ or the proanthocyanidin-containing plant tissue can become blue, wherein the higher the content of the proanthocyanidin, the bluer the color, the color of the proanthocyanidin-free plant organ or the proanthocyanidin-free plant tissue does not become blue, such that the purpose of rapid detection of the proanthocyanidin in the plant sample is achieved.
Description
Technical field
The present invention relates to field of phytochemistry, particularly relate to the rapid identification method of a kind of plant Central Plains anthocyanidin.
Background technology
Proanthocyanidin (Proathocyanidins) also is OPC, is a kind of of flavonoids.Proanthocyanidin is to remove one of the strongest anti-oxidant of free radical, can protect brain and nerve fiber, improves blood circulation, flexibly the effect of joint and young skin.Long-term edible height contains the food of proanthocyanidin, can alleviate diseases such as cardiovascular.The food that edible height contains proanthocyanidin has become the main Dietotherapy method that prevents these diseases, and consumption demand is increasing.
In the tissue and organ of live plant, not oxidized proanthocyanidin is colourless in cell, and naked eyes can't judge whether contain this material in the plant organ.For selecting to unoxidized proanthocyanidin in the live plant; The existing method that detects proanthocyanidin generally adopts high pressure lipuid chromatography (HPLC) (HPLC); This method needs expensive instrument, good experiment porch; The complicated sample pre-treatment, spended time is long and expense is higher; Therefore being unfavorable for large-scale popularization, inventing the method that contains proanthocyanidin in a kind of quick, easy, cheap detection plant, is to seek the important foundation of producing proanthocyanidin plant and large-scale development proanthocyanidin product.
At present, do not see to have and utilize the report that contains proanthocyanidin species method in the chemical dyeing method fast detecting plant based on our result of study, the present invention provides a kind of new method for whether containing proanthocyanidin in the detection plant.
Summary of the invention
The object of the present invention is to provide the method for quick of proanthocyanidin in a kind of directly perceived, economic, easy plant, can be applicable to detect the content of proanthocyanidin in the plant.The inventive method uses reagent cheap, simple to operate, and effect is obvious, instant effect.
Technical scheme of the present invention: at normal temperatures; Soak plant organ to be detected or tissue sample about 5 minutes with the acid alcohol solution of DMACA (p-dimethylaminocinnamaldehyde); Then can be if contain proanthocyanidin in plant organ or the tissue sample by colors blue; And proanthocyanidin content is high more, and color is more blue; If do not contain proanthocyanidin then not by colors blue in the sample, thereby reach the purpose that whether contains proanthocyanidin in the fast detecting plant sample.
The acid alcohol solution of said DMACA is DMACA hydrochloric acid-ethanolic solution or DMACA hydrochloric acid-methanol solution.
In described DMACA hydrochloric acid-ethanolic solution, the concentration of DMACA is 0.5-2%, and concentration of hydrochloric acid is 10-15%, and concentration of alcohol is 45-55%.
In described DMACA hydrochloric acid-methanol solution, the concentration 0.5-2% of DMACA, concentration of hydrochloric acid are 8-14%, and methanol concentration is 50-55%.
The applicant is according to the characteristics of vegetable material and the chemical property of proanthocyanidin in invention; Adopt the softening plant tissue of hydrochloric acid of high concentration in the present invention and sour environment is provided; Help coloring agent and get into cell and proanthocyanidin reaction; Proanthocyanidin content is high more in plant organ or the tissue, and color is more blue, thus can be easy, intuitively distinguish plant organ or organize in proanthocyanidin have or not the height with content.
Embodiment
Embodiment 1
(1) get proanthocyanidin standard items (replacing this Chinese mugwort institute of Chinese materia medica), be mixed with the proanthocyanidin solution of following concentration respectively available from Nanjing: 1mg/ml, 3mg/ml, 5mg/ml, get 1 milliliter of proanthocyanidin solution respectively and place transparent vessel;
(2) add 1 gram DMACA (p-dimethylaminocinnamaldehyde) in 100 milliliters of hydrochloric acid-ethanolic solutions, wherein concentration of hydrochloric acid is 10%, and concentration of alcohol is 50%, and fully dissolving gets DMACA hydrochloric acid-ethanolic solution.
(3) in the transparent vessel that proanthocyanidin solution is housed, add 1 milliliter of above-mentioned DMACA hydrochloric acid-ethanolic solution respectively; Observed color after dyeing at normal temperatures 5 minutes; All produce bluely in all solution, and blue degree increases along with the increase of proanthocyanidin concentration successively.
Embodiment 2
(1) get respectively the back 8 days arabidopsis of two kinds of dissimilar pollinations (
Arabidopsis thaliana) the kind skin of seed, one of them is the seed kind skin of wild type, contains proanthocyanidin in the bibliographical information kind skin, and another is the seed kind skin of yellow seed mutant, and bibliographical information kind Pi Zhongwei detects proanthocyanidin.
(2) add 1.5 gram DMACA (p-dimethylaminocinnamaldehyde) in 100 milliliters of hydrochloric acid-ethanolic solutions, wherein concentration of hydrochloric acid is 15%, and concentration of alcohol is 50%, and fully dissolving gets DMACA hydrochloric acid-ethanolic solution.
(3) get kind skin that above-mentioned material pollinates back 8 days respectively on microslide, cover fully with DMACA hydrochloric acid-ethanolic solution.Dyeing is after 5 minutes down for normal temperature, and the kind skin of wild type arabidopsis seed is dyed by green and is blueness, and the seed kind skin of yellow seed mutant still is green not by colors blue.
Embodiment 3:
(1) get respectively the back 10 days Chinese cabbage of two kinds of dissimilar pollinations (
Brassica rapa) the kind skin of seed, one of them contains proanthocyanidin for hiding No. 2 seed kind skins of oil in the bibliographical information kind skin, and another is the seed kind skin of Peng Bohuang, does not contain proanthocyanidin in the bibliographical information kind skin.
(2) add 1 gram DMACA (p-dimethylaminocinnamaldehyde) in 100 milliliters of hydrochloric acid-methanol solutions, wherein concentration of hydrochloric acid is 14%,, methanol concentration is 55%, fully dissolving gets DMACA hydrochloric acid-methanol solution.
(3) get seed kind skin that above-mentioned material pollinates back 10 days respectively on microslide, cover fully with DMACA hydrochloric acid-methanol solution.Dyeing is after 5 minutes down for normal temperature, and the seed kind skin of hiding oily No. 2 becomes blueness by green, and Peng Bo yellow race skin is not caught look, still is green.
Embodiment 4:
(1) get respectively the back 10 days paddy rice of two kinds of dissimilar pollinations (
Oryza.sativa) caryopsis, one of them is the red rice caryopsis, and bibliographical information wherein contains proanthocyanidin, and another is the rice caryopsis, and bibliographical information does not wherein detect proanthocyanidin.
(2) add 0.5 gram DMACA (p-dimethylaminocinnamaldehyde) in 100 milliliters of hydrochloric acid-ethanolic solutions, wherein concentration of hydrochloric acid is 10%, and concentration of alcohol is 55%, and fully dissolving gets DMACA hydrochloric acid-ethanolic solution.
(3) get the back 10 days caryopsis of the pollination of above-mentioned material respectively on microslide, cover fully with DMACA hydrochloric acid-ethanolic solution.Dyeing is after 5 minutes down for normal temperature, and the red rice caryopsis becomes blueness by green, and the rice caryopsis is not caught look, still is green.
Embodiment 5:
(1) fetch the dissimilar tealeaves in source (
Camellia sinensis) fresh leaf sample, color is green, HPLC measures wherein that the content of proanthocyanidin is followed successively by by height on earth: sample 1; Sample 2>Sample 3.
(2) add 2 gram DMACA (p-dimethylaminocinnamaldehyde) in 100 milliliters of hydrochloric acid-ethanolic solutions, wherein concentration of hydrochloric acid is 10%, and concentration of alcohol is 55%, and fully dissolving gets DMACA hydrochloric acid-ethanolic solution.
(3) get the blade of above-mentioned fresh tea leaf in its respectively, be cut into small pieces, be positioned over respectively on the microslide, cover fully with DMACA hydrochloric acid-ethanolic solution.Dyeing is after 5 minutes down for normal temperature, and sample is all by colors blue, and blue degree is followed successively by from shallow to deep: sample 3 < sample 2 < sample 1.
Claims (5)
1.
OnePlant the method that contains proanthocyanidin in the fast detecting plant; It is characterized in that, at normal temperatures, soak plant organ or tissue sample to be detected with the acid alcohol solution of DMACA; Then can be if contain proanthocyanidin in plant organ or the tissue sample by colors blue; And proanthocyanidin content is high more, and color is more blue, if do not contain proanthocyanidin then not by colors blue in the sample.
2. contain the method for proanthocyanidin in the fast detecting plant according to claim 1, it is characterized in that, the acid alcohol solution of said DMACA is DMACA hydrochloric acid-ethanolic solution.
3. contain the method for proanthocyanidin in the fast detecting plant according to claim 1, it is characterized in that, the acid alcohol solution of said DMACA is DMACA hydrochloric acid-methanol solution.
4. the method that contains proanthocyanidin in the fast detecting plant according to claim 2, it is characterized in that for, in described DMACA hydrochloric acid-ethanolic solution, the concentration of DMACA is 0.5-2%, concentration of hydrochloric acid is 10-15%, concentration of alcohol is 45-55%.
5. contain the method for proanthocyanidin in the fast detecting plant according to claim 3, it is characterized in that, in described DMACA hydrochloric acid-methanol solution, the concentration 0.5-2% of DMACA, concentration of hydrochloric acid are 8-14%, and methanol concentration is 50-55%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201110250289A CN102323378A (en) | 2011-08-29 | 2011-08-29 | Method for rapidly detecting proanthocyanidin in plant |
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| CN201110250289A CN102323378A (en) | 2011-08-29 | 2011-08-29 | Method for rapidly detecting proanthocyanidin in plant |
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Citations (9)
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|---|---|---|---|---|
| EP0790314A2 (en) * | 1996-02-14 | 1997-08-20 | Kikkoman Corporation | Process for the preparation of proanthocyanidins |
| JPH1180148A (en) * | 1997-09-02 | 1999-03-26 | Kikkoman Corp | Production of proanthocyanidin |
| CN1281857A (en) * | 2000-07-08 | 2001-01-31 | 刘征涛 | Microwave extraction method of protocyaidin-like substances from grape seed |
| JP2001247715A (en) * | 2000-03-02 | 2001-09-11 | Terumo Corp | Cellulose solution, and stabilizer for cellulose solution |
| CN1431205A (en) * | 2002-01-11 | 2003-07-23 | 华中农业大学 | Extraction agent used for extracting proanthocyanidin from plants and extracting method |
| CN1506361A (en) * | 2002-12-06 | 2004-06-23 | 沈阳中科药业公司 | Low temperature proanthocyanidin extracting process from grape seed |
| CN1556150A (en) * | 2004-01-12 | 2004-12-22 | 四川大学 | Extraction and separation of proto flower haematochrome and preparation method of its derivatives |
| CN1989407A (en) * | 2004-07-29 | 2007-06-27 | 三得利株式会社 | Method for analyzing oligomeric proanthocyanidin (opc) |
| WO2007102146A2 (en) * | 2006-03-06 | 2007-09-13 | Zetiq Technologies Ltd. | Methods and compositions for identifying a cell phenotype |
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2011
- 2011-08-29 CN CN201110250289A patent/CN102323378A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0790314A2 (en) * | 1996-02-14 | 1997-08-20 | Kikkoman Corporation | Process for the preparation of proanthocyanidins |
| JPH1180148A (en) * | 1997-09-02 | 1999-03-26 | Kikkoman Corp | Production of proanthocyanidin |
| JP2001247715A (en) * | 2000-03-02 | 2001-09-11 | Terumo Corp | Cellulose solution, and stabilizer for cellulose solution |
| CN1281857A (en) * | 2000-07-08 | 2001-01-31 | 刘征涛 | Microwave extraction method of protocyaidin-like substances from grape seed |
| CN1431205A (en) * | 2002-01-11 | 2003-07-23 | 华中农业大学 | Extraction agent used for extracting proanthocyanidin from plants and extracting method |
| CN1506361A (en) * | 2002-12-06 | 2004-06-23 | 沈阳中科药业公司 | Low temperature proanthocyanidin extracting process from grape seed |
| CN1556150A (en) * | 2004-01-12 | 2004-12-22 | 四川大学 | Extraction and separation of proto flower haematochrome and preparation method of its derivatives |
| CN1989407A (en) * | 2004-07-29 | 2007-06-27 | 三得利株式会社 | Method for analyzing oligomeric proanthocyanidin (opc) |
| WO2007102146A2 (en) * | 2006-03-06 | 2007-09-13 | Zetiq Technologies Ltd. | Methods and compositions for identifying a cell phenotype |
| WO2007102146A3 (en) * | 2006-03-06 | 2007-11-29 | Zetiq Technologies Ltd | Methods and compositions for identifying a cell phenotype |
Non-Patent Citations (8)
| Title |
|---|
| TAYLOR C.，ETC: "Evaluation of Parameters that Affect the 4-Dimethylaminocinnamaldehyde Assay for Flavanols and Proanthocyanidins", 《JOURNAL OF FOOD SCIENCE》, vol. 75, no. 7, 26 May 2010 (2010-05-26), pages 619 - 625 * |
| W. FEUCHT,ETC: "Flavanol binding of nuclei from tree species", 《PLANT CELL REPORTS》, vol. 22, no. 6, 1 November 2003 (2003-11-01), pages 430 - 436 * |
| 张洁等: "葡萄穗轴中白藜芦醇、原花色素提取工艺的研究", 《河北工业大学学报》, no. 01, 15 February 2008 (2008-02-15) * |
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Application publication date: 20120118 |