TWI546109B - Method for extracting chloropyll - Google Patents
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- TWI546109B TWI546109B TW099143528A TW99143528A TWI546109B TW I546109 B TWI546109 B TW I546109B TW 099143528 A TW099143528 A TW 099143528A TW 99143528 A TW99143528 A TW 99143528A TW I546109 B TWI546109 B TW I546109B
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Description
本發明涉及一種萃取葉綠素的方法。 The present invention relates to a method of extracting chlorophyll.
葉綠素是光合作用膜中的綠色色素,它是光合作用中捕獲光的主要成分。葉綠素共有a、b、c、d和e等幾種。凡進行光合作用時釋放氧氣的植物均含有葉綠素a;葉綠素b存在於高等植物、綠藻和眼蟲藻中;葉綠素c存在於矽藻、鞭毛藻和褐藻中,葉綠素d存在於紅藻。高等植物葉子本體中的葉綠素主要有葉綠素a和葉綠素b兩種。葉綠素a的分子結構由4個吡咯環通過4個甲烯基連接形成環狀結構,稱為卟啉。卟啉環中央結合著1個鎂原子,並有一環戊酮(V),在環IV上的丙酸被葉綠醇酯化、皂化後形成鉀鹽具水溶性。它們不溶於水,而溶於有機溶劑,如乙醇、丙酮、乙醚、氯仿等,所以可利用有機溶劑來萃取植物體中的葉綠素。 Chlorophyll is a green pigment in photosynthesis membranes, which is the main component of light captured by photosynthesis. Chlorophyll has several kinds such as a, b, c, d and e. Plants that release oxygen during photosynthesis contain chlorophyll a; chlorophyll b is found in higher plants, green algae and Euglena; chlorophyll c is found in algae, dinoflagellates and brown algae, and chlorophyll d is present in red algae. The chlorophyll in the leaves of higher plants mainly includes chlorophyll a and chlorophyll b. The molecular structure of chlorophyll a is formed by four pyrrole rings through four methylene groups to form a cyclic structure called porphyrin. The center of the porphyrin ring is bonded to one magnesium atom and has a cyclopentanone (V). The propionic acid on the ring IV is esterified with chlorophyll, and the potassium salt formed after saponification is water-soluble. They are insoluble in water and soluble in organic solvents such as ethanol, acetone, diethyl ether, chloroform, etc., so organic solvents can be used to extract chlorophyll from plants.
本發明的目的是提供一種萃取葉綠素的方法。 It is an object of the present invention to provide a method of extracting chlorophyll.
本發明實施例提供一種萃取葉綠素的方法,所述方法包括如下步驟:步驟一:將植物葉子本體加入液態氮一起粉碎,該植物葉子本體粉碎大小為0.7毫米; 步驟二:通過有機溶劑萃取葉綠素;步驟三:將萃取好葉綠素的液體,由濾紙過濾,然後利用濃縮機濃縮液體;步驟四:收集濃縮好的液體。 An embodiment of the present invention provides a method for extracting chlorophyll, the method comprising the following steps: Step 1: pulverizing the plant leaf body with liquid nitrogen, the plant body body pulverizing size is 0.7 mm; Step 2: extracting chlorophyll by an organic solvent; Step 3: extracting the chlorophyll-containing liquid, filtering it by a filter paper, and then concentrating the liquid by a thickener; Step 4: collecting the concentrated liquid.
優選地,在所述步驟一中,粉碎植物葉子本體之前,先將植物葉子本體先低溫保存,並且進行曬乾以蒸發植物葉子本體內的水份。 Preferably, in the first step, before the body of the plant leaves is pulverized, the plant leaf body is firstly stored at a low temperature, and dried to evaporate the water in the body of the plant leaf.
優選地,在所述步驟一的植物葉子本體粉碎過程中,加入液態氮與植物葉子本體一起粉碎。 Preferably, in the step of pulverizing the plant leaves in the first step, liquid nitrogen is added and pulverized together with the plant leaf body.
優選地,所述有機溶劑包括乙醇、丙酮、二甲基甲醯胺及二甲基亞碸中的一種或多種。 Preferably, the organic solvent includes one or more of ethanol, acetone, dimethylformamide, and dimethylhydrazine.
優選地,在所述步驟二中,將萃取液置於暗室環境中完成,並利用離心機進行萃取。 Preferably, in the second step, the extract is placed in a dark room environment and extracted using a centrifuge.
優選地,在所述步驟三中,所述濾紙為選自定性濾紙、定量濾紙、有機溶劑定量濾紙及玻璃纖維濾紙中的一種或多種。 Preferably, in the third step, the filter paper is one or more selected from the group consisting of a qualitative filter paper, a quantitative filter paper, an organic solvent quantitative filter paper, and a glass fiber filter paper.
優選地,在所述步驟三中,控制過濾程式的時間,使得在24小時內完成過濾程式。 Preferably, in the third step, the time of the filtering program is controlled so that the filtering program is completed within 24 hours.
優選地,在所述步驟四中,將收集好的濃縮液體放置於冷藏櫃保存。 Preferably, in the fourth step, the collected concentrated liquid is placed in a refrigerator for storage.
優選地,在所述步驟四中,將冷藏櫃的溫度設為4攝氏度。 Preferably, in the fourth step, the temperature of the refrigerator is set to 4 degrees Celsius.
包括附圖以提供對於本發明的進一步理解,且附圖併入本說明書中並且構成本說明書的一部份。附圖說明本發明之示範性實施例。在諸圖中:圖1是本發明較佳實施例中植物葉子本體保存的流程圖;圖2是本發明較佳實施例中萃取和檢測葉綠素的流程圖;圖3是三種有機溶液吸收光譜對比圖。 The drawings are included to provide a further understanding of the invention, and are incorporated in this specification and constitute a part of this specification. BRIEF DESCRIPTION OF THE DRAWINGS Exemplary embodiments of the invention. In the drawings: Figure 1 is a flow chart of the preservation of plant leaves in a preferred embodiment of the present invention; Figure 2 is a flow chart for extracting and detecting chlorophyll in a preferred embodiment of the present invention; Figure 3 is a comparison of absorption spectra of three organic solutions. Figure.
下面結合附圖和實施例對本發明實施例進行詳細說明。 The embodiments of the present invention are described in detail below with reference to the accompanying drawings and embodiments.
表1是自然界中的幾種葉綠素種類列表圖。葉綠素是存在植物、藻類和藍藻類中的光合色素,其中色素為二氫卟吩(Chlorin)。一般在生活中所常見的植物普遍存在葉綠素a和葉綠素b,在本發明實施例中,選定陸生植物為萃取物件,具體步驟如下所述。 Table 1 is a list of several chlorophyll species in nature. Chlorophyll is a photosynthetic pigment found in plants, algae and cyanobacteria, wherein the pigment is Chlorin. Chlorophyll a and chlorophyll b are ubiquitous in plants commonly found in life. In the present embodiment, terrestrial plants are selected as extracting articles, and the specific steps are as follows.
圖1是本發明較佳實施例中植物葉子本體保存的流程圖。如圖1所 示,植物葉子本體摘除後,先用低溫的方式做保鮮的工作,最後將植物葉子本體曬乾以將多餘的水分蒸發。 BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a flow chart showing the preservation of a plant leaf body in a preferred embodiment of the present invention. As shown in Figure 1 It is indicated that after the leaves of the plant are removed, the work of preserving the fresh water is first carried out, and finally the leaves of the plants are dried to evaporate excess water.
圖2是本發明較佳實施例中萃取葉綠素的流程圖。如圖2所示,在步驟一S1中,進行植物葉子本體的粉碎動作。首先獲取植物葉子本體並進行圖1所示的預處理。用液態氮浸泡經過步驟一處理後的植物葉子本體,然後利用粉碎機將植物葉子本體磨成大小為0.7毫米(mm)的細粉末。萃取分析前的細粉末需放置於黑暗且低溫的條件下儲藏。粉碎機可採用一般食品用的果汁機及製粉機,但是在粉碎的過程中會有溫度的問題,影響到實驗,所以在實驗的粉碎過程中加入液態氮與植物葉子本體一起粉碎。 Figure 2 is a flow diagram of the extraction of chlorophyll in a preferred embodiment of the invention. As shown in Fig. 2, in step S1, the pulverizing action of the plant leaf body is performed. The plant leaf body is first obtained and the pretreatment shown in Figure 1 is performed. The body of the plant leaves subjected to the step 1 treatment was soaked with liquid nitrogen, and then the body of the plant leaves was ground into a fine powder having a size of 0.7 mm by means of a pulverizer. The fine powder before the extraction analysis should be stored in a dark and low temperature condition. The pulverizer can use a juice machine and a pulverizer for general foods, but there is a temperature problem during the pulverization process, which affects the experiment. Therefore, liquid nitrogen is added to the body of the plant to be pulverized in the pulverization process of the experiment.
在步驟二S2中,進行葉綠素的溶液萃取動作。在萃取葉綠素的過程中,可使用以下溶液的一種或多種:乙醇、丙酮、二甲基二醯胺(DMF)及二甲亞碸(DMSO),將上述的各種溶液分別加入磨成的細粉末,然後利用離心機萃取葉綠素,在萃取時,實驗條件優選為暗室。 In step two S2, a solution extraction operation of chlorophyll is performed. In the process of extracting chlorophyll, one or more of the following solutions may be used: ethanol, acetone, dimethyldiamine (DMF) and dimethyl hydrazine (DMSO), and the above various solutions are separately added to the ground fine powder. Then, the chlorophyll is extracted by a centrifuge, and in the extraction, the experimental condition is preferably a dark room.
乙醇及丙酮這二種溶液較容易揮發,在實驗上非常方便,二甲基二醯胺及二甲亞碸對人體傷害較大,在做二甲基二醯胺及二甲亞碸萃取時需注意揮發的氣體,所以需要額外的設備,使溶液氣體排出,主要利用真空烘箱以及排氣櫃,利用排氣櫃將揮發之氣體排出。如果需要加速揮發氣體,也可利用真空烘箱,用加熱及抽真空的方式,加速氣體排出。 The two solutions of ethanol and acetone are relatively easy to volatilize, and it is very convenient in experiment. Dimethyldiamine and dimethyl hydrazine are harmful to the human body, and it is needed for the extraction of dimethyldiamine and dimethyl hydrazine. Pay attention to the volatilized gas, so additional equipment is needed to discharge the solution gas. The vacuum oven and the exhaust cabinet are mainly used, and the volatilized gas is discharged by the exhaust cabinet. If it is necessary to accelerate the volatilization gas, the vacuum oven can also be used to accelerate the gas discharge by heating and vacuuming.
在步驟三S3中,進行葉綠素的過濾、濃縮動作。將萃取好的液體,優選在24小時內,以篩檢程式將植物葉子本體細粉末濾除,主要經由濾紙濾除不要的部分。不同的濾紙有不同的效果,大致可 分為定性濾紙、定量濾紙、有機溶液定量濾紙及玻璃纖維濾紙。可根據不同的需要挑選濾紙。 In the third step S3, the chlorophyll filtration and concentration operation are performed. The extracted liquid, preferably within 24 hours, is filtered by a screening procedure to remove fine particles of the plant leaf body, mainly through filter paper to filter out unwanted portions. Different filter papers have different effects, roughly Divided into qualitative filter paper, quantitative filter paper, organic solution quantitative filter paper and glass fiber filter paper. Filter paper can be selected according to different needs.
濾除好的液體,可以利用濃縮機,將液體濃縮,但注意要避免過度抽乾。 To filter out good liquids, use a thickener to concentrate the liquid, but be careful to avoid excessive draining.
在步驟四S4中,進行保存動作。濾除好的溶液需放置在冷藏櫃中,最佳溫度為4攝氏度。濾除好的溶液應儘快完成葉綠素a及葉綠素b的檢測,無法立即檢測時,可將溶液放置於零下20攝氏度暗冷藏,最長可保存14天;放置於零下70攝氏度暗冷藏,最長可保存三個月。 In step S4, a save operation is performed. The filtered solution should be placed in a freezer at an optimum temperature of 4 degrees Celsius. The chlorophyll a and chlorophyll b should be detected as soon as possible. If the solution cannot be detected immediately, the solution can be stored at minus 20 degrees Celsius and stored for up to 14 days. It can be stored at minus 70 degrees Celsius and can be stored for up to three times. Months.
可利用可見光光譜儀來檢測用不同溶液中萃取的葉綠素a、葉綠素b波長的吸收強度。圖3是三種溶液吸收光譜對比圖,表2是不同有機溶液中葉綠素濃度計算結果圖。其中a為DMF溶液,b為丙酮溶液,c為DMSO溶液。由圖可知,對於從菠菜中提取的葉綠素而言,662nm附近為葉綠素a的特性吸收峰值,455nm為葉綠素b特性吸收峰值,利用已知公式分別可計算葉綠素a濃度、葉綠素b濃度與葉綠素總濃度,DMF溶液在三種溶液中萃取葉綠素的濃度含量最大,葉綠素萃取率最高。 A visible light spectrometer can be used to detect the absorption intensity of the chlorophyll a and chlorophyll b wavelengths extracted with different solutions. Figure 3 is a comparison chart of absorption spectra of three solutions, and Table 2 is a graph showing calculation results of chlorophyll concentration in different organic solutions. Where a is a DMF solution, b is an acetone solution, and c is a DMSO solution. As can be seen from the figure, for the chlorophyll extracted from spinach, the characteristic absorption peak of chlorophyll a is near 662 nm, and the peak of absorption of chlorophyll b characteristic is 455 nm. The chlorophyll a concentration, chlorophyll b concentration and total chlorophyll concentration can be calculated by known formulas. The concentration of chlorophyll in the DMF solution was the highest in the three solutions, and the chlorophyll extraction rate was the highest.
本發明的實施例主要針對電池前置作業的部分,萃取製造電池時 需要用到的葉綠素,但利用葉綠素製成電池後,可以利用儀器檢測,作用後電池中葉綠素變化。例如,PH/ORP(氧化還原電位)值的量測,檢測電池使用的過程中,是否產生有毒物質化合物。 Embodiments of the present invention are mainly directed to a portion of a battery front operation, when extracting and manufacturing a battery The chlorophyll is needed, but after the battery is made of chlorophyll, it can be detected by the instrument, and the chlorophyll changes in the battery after the action. For example, the measurement of the PH/ORP (oxidation reduction potential) value detects whether a toxic substance compound is generated during the use of the battery.
以上僅為本發明的優選實施案例而已,並不用於限制本發明,對於本領域的技術人員來說,本發明可以有各種更改和變化。凡在本發明的精神和原則之內,所作的任何修改、等同替換、改進等,均應包含在本發明的保護範圍之內。 The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes can be made to the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and scope of the present invention are intended to be included within the scope of the present invention.
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