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Method for detecting content of poa pratensis fatty acid

A technology of bluegrass and determination method, which is applied in the field of determination of fatty acid content, can solve the problems of high planting management requirements, high water content, susceptibility to diseases, etc., achieve the effects of simplifying operation steps, overcoming serious carbonization, and simplifying analysis steps

Inactive Publication Date: 2012-10-03
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, bluegrass grass is not suitable for high temperature and drought conditions, and is susceptible to diseases under high temperature conditions in northern summer, so this grass species has high requirements for planting management and requires high water, nutrient and other inputs

Method used

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  • Method for detecting content of poa pratensis fatty acid
  • Method for detecting content of poa pratensis fatty acid
  • Method for detecting content of poa pratensis fatty acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Bluegrass Grass Fatty Acid Methyl Esterification Treatment

[0046] 1. Preliminary preparation for lipid methylation

[0047] Solution reagent preparation: dilute sulfuric acid with methanol to 1N (1mol / L H + )H 2 SO 4 . Prepare 0.9% NaCl solution. Prepare chromatographic grade n-hexane and heptadecanoic acid (17:0) for use.

[0048] 2. Lipid methylation process

[0049] Take 0.2g of fresh leaf tissue of bluegrass grass, quickly cut the leaves to 1cm long sections with scissors, and put them into a test glass bottle with a lid. Add 1 mL of 1N H 2 SO 4 , containing 100 μg of heptadecanoic acid (17:0) as an internal reference, blown with nitrogen 2 Blow gently to drive out the air and quickly tighten the cap. 80°C water bath, heating for 90min to complete the fatty acid methyl esterification process.

[0050] 3. Post-treatment of lipid methylation

[0051] Place in -20°C refrigerator to cool for 10 minutes. Add 1.5 mL of 0.9% NaCl and 200 μL of n-he...

Embodiment 2

[0052] Example 2 Bluegrass Grass Fatty Acid Methyl Esterification Treatment

[0053] 1. Preliminary preparation for lipid methylation

[0054] Solution reagent preparation: dilute sulfuric acid with methanol to 1N (1mol / L H + )H 2 SO 4 . Prepare 0.9% NaCl solution. Prepare chromatographic grade n-hexane and heptadecanoic acid (17:0) for use.

[0055] 2. Lipid methylation process

[0056] Take 0.2g of fresh leaf tissue of bluegrass, quickly cut the leaves to 0.8cm long sections with scissors, and put them into a test glass bottle with a cover. Add 1 mL of 1N H 2 SO 4 , containing 100 μg of heptadecanoic acid (17:0) as an internal reference, blown with nitrogen 2 Blow gently to drive out the air and quickly tighten the cap. 60°C metal bath, heating for 120min. Complete fatty acid methylation process.

[0057] 3. Post-treatment of lipid methylation

[0058] Place in -20°C refrigerator to cool for 15 minutes. Add 1.5 mL of 0.9% NaCl and 200 μL of n-hexane (Hexane, Sigm...

Embodiment 3

[0059] Example 3 GC-MS Analysis of Bluegrass Fatty Acid

[0060] 1. Lipid content analysis:

[0061] The sample processed in Example 1 was used gas chromatography-mass spectrometer HP GG-MS (HP6890GC and HP 5973MS), using a 60-mHP-5MS capillary column with an inner diameter of 0.25mm, and the program parameters of the gas chromatograph were set as follows: 170°C 10min, rise to 220℃ for 10min, gas phase flow rate 1mL min –1 . After injecting the sample into the syringe, start the program. The lipid analysis duration for 1 sample is about 40 minutes.

[0062] 2. Obtain the peak diagram of each sample through gas phase mass spectrometry analysis. The content percentage of other fatty acids was calculated using the internal reference method, i.e. the weight of heptadecanoic acid 17:0 and the peak area on the mass spectrometry chart.

[0063] The main types of fatty acids in bluegrass tallow are as follows:

[0064] 16:0 palmitic acid, saturated fatty acid;

[0065] 16:1 pal...

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Abstract

The invention provides a method for detecting the content of poa pratensis fatty acid. The method comprises the following steps of: (1) taking poa pratensis leaf tissues in a glass bottle; (2) adding sulfuric acid containing daturic acid; (3) discharging air in the glass bottle and sealing to perform water bath or metal bath; (4) cooling, and adding sodium chloride solution and normal hexane to whirl and centrifuge; (5) taking the centrifuged upper oil phase and putting into a gas chromatography-mass spectrometer (GC-MS) to analyze, and calculating the content of fatty acid according to the sample peak diagram. The method provided by the invention overcomes the problem of inconsistent extracting efficiency of lipids among the samples, optimizes the plant tissue sample amount of the sample to be detected and saves the dosage of a series of reagent in the process of methyl esterification; furthermore, the operation step of methyl esterification is simplified, the detecting and analyzing time of GC-MS after sampling is saved; the problem of serious carbonization in the process of the methyl esterification of the poa pratensis is overcome; therefore, the method provided by the invention is a poa pratensis fatty acid analyzing method with low cost, fast speed, simplicity and high efficiency.

Description

technical field [0001] The invention relates to a method for determining fatty acid content, in particular to a method for determining fatty acid content of bluegrass grass. Background technique [0002] At present, the methods and technologies for determining the fatty acid content of plants are mostly focused on oil crops and seed parts in plant tissues. Regarding the determination method of fatty acid content in plant leaf tissue, there are relatively mature methods for the determination of fatty acid content in leaf tissue such as Arabidopsis thaliana and spinach abroad. These methods are generally divided into three main steps of oil extraction, saponification and methyl esterification, which require many kinds of reagents, complicated steps and a long time. [0003] Bluegrass (Poa Pretensis) is a perennial herb with fibrous roots, rhizomes, green leaves, good ornamental effect, rhizome reproduction, strong regeneration, pruning resistance, strong cold resistance, slig...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 许立新韩烈保黄炳茹
Owner BEIJING FORESTRY UNIVERSITY
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