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Method for quickly preparing high-purity chlorophyll and chlorophyll degradation products

A technology of chlorophyll degradation and chlorophyll, applied in the interdisciplinary field of chemistry and enzymology, can solve the problems of high experimental cost, large amount of standard products, long delivery period, etc., and achieve the effects of wide application prospects, efficient and rapid preparation, and easy operation

Active Publication Date: 2016-08-31
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Imported chlorophyll standard products often have a long delivery period (one week to one month) and are expensive (take the imported brand sigma chlorophyll a as an example, the price per mg is 6000-8000RMB), the packaging and transportation process is easy to see photodegradation, low temperature Preservation causes transportation inconvenience and other problems
Due to the above-mentioned inconvenience in the purchase process of imported standard substances, and the large amount of standard substances used when analyzing chlorophyll in samples, the cost of the experiment is very high, so the laboratory urgently needs a rapid preparation method for chlorophyll and its degradation product standard substances. However, there is currently no no reports

Method used

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  • Method for quickly preparing high-purity chlorophyll and chlorophyll degradation products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Purification and preparation of chlorophyll a and chlorophyll b

[0031] (1) Material preparation: pick the fresh sweet potato leaves purchased in the market, remove the old leaves and yellow leaves and remove the petioles, wash and dry in the shade for later use. Wash the fresh Wenzhou tangerines purchased in the market (color difference index CI is -2.0~0.0) with clear water, dry them, and set aside;

[0032] (2) Extraction of total chlorophyll (the following steps are carried out under dark conditions):

[0033] a. Weigh 100 g of sweet potato leaves washed and dried in (1), freeze with liquid nitrogen, and grind into fine powder in a mortar or sample breaker (IKA, Germany);

[0034] b. Pour the sample powder into a 500mL centrifuge bottle (Beckman), add 200mL extract solution (N,N-dimethylformamide, containing 0.1% magnesium carbonate, w / v) and mix well. Ultrasonic cleaner 100% power (100W), ice bath ultrasonic 20min, during which the sample was mixed up and down s...

Embodiment 2

[0040] Preparation of degradation products of chlorophyll a and chlorophyll b

[0041] (1) Preparation of pheophytin a and pheophytin b (pheophytin a and pheophytin b)

[0042] (a) Preparation of pheophytin a (pheophytin a): Weigh 2 mg of chlorophyll a obtained by separating and drying in step (3) of Example 1, and dissolve it in a 10 mL centrifuge tube with 4 mL of anhydrous ether to prepare 500 μg / mL. Add 13% hydrochloric acid drop by drop for acidification, fully shake each drop of hydrochloric acid until the solution turns black and stop immediately. Add 4mL double distilled water to the centrifuge tube, shake for 10 seconds, let stand to separate layers, and absorb the lower aqueous phase. Continue to add 4mL of double distilled water, oscillate and stand for stratification, absorb the lower water phase, repeat this step several times, and stop adding double distilled water when the pH of the lower water phase is measured with pH test paper at 6.8-7.2. Centrifuge at 400...

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Abstract

The invention discloses a method for quickly preparing high-purity chlorophyll and chlorophyll degradation products. Satsuma mandarin peels and sweet potato leaves are used as raw materials for the first time to prepare the chlorophyll and derivatives thereof. The method disclosed by the invention is convenient and quick for taking materials, low in cost and easy to operate, and can complete the preparation of all the 8 products within three days. The purity of the obtained products can reach 90-95%, and the cost is much lower than the price of the same type of imported standard substance. The method can efficiently and quickly prepare the standard reference substances of the milligram-level chlorophyll and derivatives thereof required by the experiment, is suitable for popularization and application in commercial laboratories and enterprises, and has wide application prospects.

Description

technical field [0001] The invention belongs to the cross field of chemistry and enzymology, in particular to a method for rapidly preparing high-purity chlorophyll and chlorophyll degradation products. Background technique [0002] At present, when laboratories analyze chlorophyll and its degradation products, they usually use imported brand standard reference substances as reference. These standard reference substances are usually purified from materials such as spinach leaves and green algae by preparative thin-layer chromatography (TLC) and preparative HPLC. The purity is usually 90%-98%. Usually, the higher the purity, the more expensive it is. Imported chlorophyll standard products often have a long delivery period (one week to one month) and are expensive (take the imported brand sigma chlorophyll a as an example, the price per mg is 6000-8000RMB), the packaging and transportation process is easy to see photodegradation, low temperature Preservation causes problems s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D487/22C12P17/18
CPCC07D487/22C12P17/182
Inventor 程运江罗焘瞿韵李卓然刘欢
Owner HUAZHONG AGRI UNIV
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