How to Extract and Determine Chlorophyll of Lotus Leaf
The lotus leaf, also known as the lotus leaf and the loquat leaf, is the leaf of the lotus, a perennial, large-scale aquatic herb of the genus Nymphaea. Lotus root vegetables, which are not required for growth conditions, are distributed in the north and south of China and are mainly distributed in the Yangtze River basin and the southern provinces. Studies have shown that the lotus leaf is rich in nutrients, and in addition to common chemical components such as carbohydrates, lipids, proteins, and tannin that are common to common plants, it is also rich in flavonoids and alkaloids. Chlorophyll as a natural pigment is widely used in medicine, food, cosmetics and other industries. With the improvement of people's living standards, the demand for natural pigments is also growing. Chlorophyll determination is mainly measured by a chlorophyll meter.
This article mainly analyzes the extraction and determination of chlorophyll from lotus leaf. The chlorophyll extraction process of the lotus leaf is: lotus leaf→washing→dried surface moisture→cutting→weighing→grinding→leaching→centrifugation→absorbance measurement. The specific operation for extraction is: selection of the extraction solvent. Weigh 5g of lotus leaf, using ether, ethanol, formamide, methanol, acetone, chloroform, dimethyl sulfoxide as the extractant, and the ratio of solid to liquid is 1:50, leaching at room temperature for 1 day respectively, collecting and extracting. After the liquid was brought to a constant volume, the absorbance was measured at a wavelength of 652 nm. As can be seen from the results, with the exception of chloroform, the extraction rates of other extractants on the chlorophyll of the lotus leaf were all higher, and the highest was acetone. Therefore, acetone was used as an extraction solvent. 2. Selection of extractant concentration. Weigh 5g of lotus leaves, respectively 65%, 75%, 80%, 85%, 90%, 95% acetone and anhydrous acetone as the extractant, the ratio of material to liquid 1:10, heated to reflux at 50 ° C for 1.5h. After determining the volume, the absorbance was measured at a wavelength of 652 nm. The results showed that when the acetone concentration was 80% to 90%, the extraction effect was good. 3. The ratio of liquid to liquid. Weigh 5g of lotus leaf, take 85% acetone as the extractant, and the ratio of material to liquid is 1:5, 1:6, 1:7, 1:8, 1:9. Heat and reflux at 50°C for 1.5 h. Absorbance was measured at 652 nm. It can be seen that when the ratio of solid to liquid is 1:7, the extraction effect is obviously better than other conditions. 4. The choice of extraction temperature. Weigh 5g of lotus leaf, with the selected solvent, material to liquid ratio, respectively, at 30 °C, 40 °C, 50 °C, 60 °C, 70 °C, 80 °C, 90 °C heated reflux extraction 1.5h, the extract was constant volume, The absorbance was measured at a wavelength of 652 nm. It can be seen that the extraction effect of chlorophyll in the low temperature stage is good, and the absorbance of the extract at 30°C to 40°C is significantly increased, and is constant at 40°C to 60°C, but when the temperature exceeds 60 After °C, the absorbance of the extract began to drop, which may be related to the thermal stability of chlorophyll itself. 50°C was selected as the temperature condition for further study. 5. The choice of extraction time. Weigh the lotus leaf 5g, immerse at 50 °C for 0.5h, 1.0h, 1.5h, 2.0h, 2.5h with the selected solvent, liquid-to-liquid ratio, and measure the absorbance at a wavelength of 652 nm after constant volume of the extract. Values, the results can be seen, the extraction time is 1.5h, the best extraction.
According to the extraction conditions of lotus leaf chlorophyll, the optimum extraction process of lotus leaf chlorophyll was that 85% acetone was used as extractant, the ratio of material to liquid was 1:8, the extraction temperature was 50 °C, and the extraction time was 1.5 h. The value of the chlorophyll absorbance of the lotus leaf under the optimized process was 2.249. After the chlorophyll has been extracted, we can determine the chlorophyll content, which includes chemical methods and relatively simple instrument methods. Now generally, if not precisely, we generally use chlorophyll meters to measure, because the chlorophyll meter is quick and easy to display chlorophyll SPAD values ​​in 2 seconds.
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