Solvent-extraction-and-fluorescence determination method of microalgae lipid content

Abstract

The invention discloses a solvent-extraction-and-fluorescence determination method of microalgae lipid content, and belongs to the technical field of microalgae crude lipid determination. The solvent-extraction-and-fluorescence determination method of the microalgae lipid content solves the problems of a fluorimetry in the prior art that dyeing with direct use of Nile red fluorescent staining is halfway and disturbance of microalgae body chlorophyll fluorescence peaks and the like occur when an ultrasound method or a dimethyl sulfoxide (DMSO)-processing-Nile-red-dyeing fluorescence determination method is in use. The solvent-extraction-and-fluorescence determination method of microalgae lipid content comprises the following steps of preparing a standard curve by using microalgae to prepare standard microalgae liquid, extracting microalgae lipid and determining content of the microalgae lipid by using fluorescence. Due to the fact that in a determination process, operation steps are free from evaporation organics, and pollution to the environment is little. The solvent-extraction-and-fluorescence determination method of the microalgae lipid content has the advantages that the solvent-extraction-and-fluorescence determination method of the microalgae lipid content is rapid, simple, effective and sensitive, and determination results are free from influence of retention periods of microalgae samples.

Description

technical field [0001] The invention belongs to the technical field of measuring microalgae crude oil, in particular to a solvent extraction-fluorescence method for measuring the content of microalgae oil. Background technique [0002] With the development of the economy, human beings have higher and higher requirements for energy, and microalgae are considered to be ideal raw materials for the preparation of biodiesel fuel. Algae has the advantages of high photosynthetic efficiency, zero net carbon value, easy cultivation, short growth cycle, high oil hydrocarbon content, etc., and does not compete with farmers for land, does not compete with people for food, and does not affect global energy distribution. As a bioenergy source, it has caused Governments and scholars of various countries pay close attention to it. The research process of producing biodiesel from microalgae includes screening of oil-producing microalgae, microalgae culture system, collection and separation ...

AI Technical Summary

Problems solved by technology

[0004] Although the organic solvent extraction method is widely used, easy to operate, and low in cost, it requires a large amount of sample, and the obtained total lipid content is low. When the amount of microalgae is relatively small, the oil cannot be extracted or the extracted oil cannot be weighed. It is easy to volatilize when removing organic solvents, the volatile solvents are toxic and easy to pollute the environment, and the operation is time-consuming and laborious; the Soxhlet extraction method is more accurate in determining the total lipid content of microalgae. Volatile and pollute the environment, and the extraction time is long; fluorescent dye determination method, Fourier transform infrared spectroscopy, vanillin phosphate method, copper reagent method, Sudan black staining method and nuclear magnetic resonance method require less sample amount and shorter time , but there are still deficiencies

Among them, the fluorescent dye method does not need to extract oil and has high sensitivity, but it is easily affected by various factors such as incomplete dyeing and chlorophyll fluorescence peaks; Fourier transform infrared spectroscopy and nuclear magnetic resonance methods do not damage the sample and do not require or Only a little pretreatment is required, the latter has good repeatability, but the cost is high; the vanillin phosphoric acid assay is relatively stable, but it needs to use a strong acid such as sulfuric acid, and the operation is more dangerous; the copper reagent method is low in cost, but it is rarely reported. less; the Sudan black staining method is simple to operate, does not need to break the algae cells and chemical extraction, but it is not suitable for the determination of non-uniform samples, and is affected by the growth and metabolism of algal cells, so it cannot be stored. If the algae samples cannot be used, this method can be used for determination Ineffective, and the measurement technology is not mature enough, currently it is mostly used for fat staining in medical treatment; supercritical CO 2 The extraction method and the ionic liquid extraction method have high extraction efficiency, and the obtained total lipid content is relatively large, but the determination cost is relatively high

[0005] At present, the most commonly used determination method is the fluorescence measurement method. The fluorescence measurement method directly uses the Nile red fluorescence staining method to stain thoroughly, but the method of ultrasonic or dimethyl sulfoxide (DMSO) treatment-Nile red staining to measure the fluorescence can overcome the problem. The dyeing is not thorough enough, but the ultrasonic or DMSO treatment-fluorescence method measures the algae. When the algae are cultivated for a certain period of time, they need to be measured in time and cannot be stored for a long time. If the experiment fails, the algae need to be re-raised; and the extracted Algae oil can be stored, and if the experimental measurement fails, it can also be reconfigured for measurement

Furthermore, when the concentration of the algae liquid is high, directly measuring the fluorescence with the algae body is easily interfered by the chlorophyll fluorescence peak. At the same time, the chlorophyll content of different algae samples is different, and it is more affected by the chlorophyll fluorescence peak.