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Method for measuring starch content of oceanic microalgae cell

A technology of starch content and marine microalgae, which is applied in the preparation of test samples, material analysis by observing the impact on chemical indicators, and analysis by making materials undergo chemical reactions can solve the problems of incomplete starch extraction and experimental problems. Long cycle, expensive and other issues, to achieve the effect of long color stability time, short experiment cycle, and simple process

Inactive Publication Date: 2012-01-18
DALIAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The former mainly uses water as the starch extractant, the starch extraction is not thorough, and the interfering substances are difficult to remove; the latter is a method for rapid analysis and determination of starch proposed by the international Swedish J.Holm [1] , the method is accurate, but the samples need to be treated with α-amylase, amyloglucosidase, and glucose oxidase successively during the experiment. It is difficult to purchase the three enzymes at the same time in China, and the price is expensive.
The anthrone colorimetric method and enzymatic hydrolysis method (GB 5009.9-85) are mostly used in China, both of which first extract and convert the starch in the plant into reducing sugar, and then calculate the starch content indirectly by quantitatively measuring the reducing sugar content. Reducing sugar is not easy to remove and affects the accuracy of the results. The experimental cycle is long and the procedures are complicated. These unfavorable factors bring a lot of inconvenience to the laboratory analysis operation.

Method used

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  • Method for measuring starch content of oceanic microalgae cell
  • Method for measuring starch content of oceanic microalgae cell
  • Method for measuring starch content of oceanic microalgae cell

Examples

Experimental program
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Effect test

Embodiment 1

[0026] (1) Drawing of starch standard curve

[0027] Accurately weigh about 0.10g of analytically pure starch, dissolve it in boiled deionized water, and after cooling, set the volume to a 1L volumetric flask to prepare a starch standard solution; then measure different volumes of the starch standard solution, and use 0.8g / mL calcium nitrate solution was added to 4ml, add 0.15mL 0.01mol / L I 2 -KI standard solution, shake well, utilize T6 new century ultraviolet-visible spectrophotometer to measure its absorbance value under 570nm wavelength respectively, add 0.15mL 0.01mol / L I to 0.8g / mL calcium nitrate solution of 4ml 2 -KI standard solution is used as a blank, the results are shown in the following table 1, and the volume of the starch standard solution is plotted with absorbance, see the appendix figure 1 .

[0028] Table 1 The absorbance values ​​of different volume standard starch solutions

[0029]

[0030]

[0031] Such as figure 1 As shown, the abscissa is t...

Embodiment 2

[0054] Determination of starch content in microalgae samples

[0055] A. Centrifuge the marine microalgae culture solution cultivated with seawater to the equilibrium period at 3200 rpm for 7 minutes to obtain fresh microalgae mud;

[0056] B. Wash the fresh microalgae mud with 1 times of deionized water repeatedly for 3 times, remove the impurities in the reaction system and change the osmotic pressure inside and outside the cells;

[0057] C. After repeated freezing and thawing of the microalgae mud washed with deionized water for 5 times, ultrasonically treat it for 15 minutes to break the microalgae cells;

[0058] D. Weigh 0.5231g, 0.5179g, 0.5082g of crushed microalgal algae mud respectively, mix them with 3ml of absolute ethanol, shake for 3min, then centrifuge at 3000rpm for 5min, discard the supernatant, and keep the precipitate.

[0059] E. Add 2ml of petroleum ether to the precipitate, mix and shake for 5 minutes, then add 3ml of 90% ethanol, mix and shake for 5 mi...

Embodiment 3

[0066] Determination of starch content in microalgae samples

[0067] A. Centrifuge the marine microalgae culture solution cultivated with seawater to the equilibrium period at 3500rpm for 5min to obtain fresh microalgae slime;

[0068] B. Wash the fresh microalgae mud with 1 times of deionized water repeatedly for 3 times, remove the impurities in the reaction system and change the osmotic pressure inside and outside the cells;

[0069] C. After repeated freezing and thawing of the microalgae mud washed with deionized water for 5 times, ultrasonically treat it for 15 minutes to break the microalgae cells;

[0070] D. Weigh 0.5033g, 0.5264g, 0.5096g of crushed microalgae slime, mix with 3ml of absolute ethanol, shake for 3min, centrifuge at 3000rpm for 5min, discard the supernatant, and keep the precipitate.

[0071] E. Add 2ml of petroleum ether to the precipitate, mix and shake for 5 minutes, then add 3ml of 90% ethanol, mix and shake for 5 minutes, centrifuge at 3000rpm fo...

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Abstract

The invention discloses a method for measuring starch content of an oceanic microalgae cell. In allusion to features of the substance composition of the oceanic microalgae cell, the method comprises the following steps of: firstly, degreasing, decolorizing and desugaring the microalgae cell by using ethanol, petroleum ether and 90% ethanol; secondly, taking a calcium nitrate solution with the concentration of 0.6-0.8g / ml as an extractant for extracting starch in the microalgae cell, and boiling for 15 to 30 minutes to sufficiently extract the starch in the microalgae cell; thirdly, oxidizing residual reducing sugar and residual pigment by using hydrogen peroxide, and boiling to remove the excess hydrogen peroxide; and finally, taking iodine as a color-developing agent, and directly measuring the starch content of the oceanic microalgae through spectrophotometry. Compared with the conventional method for measuring starch content of terrestrial plants, the method has the advantages of simple process, short experimental period, accurate result and good repeatability.

Description

technical field [0001] The invention belongs to the field of marine bioenergy, and in particular relates to a method for measuring starch content in marine microalgae cells. Background technique [0002] Microalgae have high photosynthetic utilization efficiency, small individuals, rich nutrition, rapid growth and reproduction, strong adaptability to the environment, easy cultivation, and do not occupy agricultural land. Many achievements and developments. Especially in recent years, the development and application of biomass energy using marine microalgae has attracted the attention of many researchers. Starch in microalgae cells can be degraded, fermented, and finally converted into energy that can be directly used. An important problem that needs to be solved first is how to screen starch-rich microalgae and how to increase starch content by optimizing the culture of microalgae , the accurate determination of starch content is the key. At present, there is no report on...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N1/28G01N1/38
Inventor 徐嫔杨海波孟迎迎于媛
Owner DALIAN UNIV
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