Food lipid extraction method and detection method of lipid in food

A food and lipid technology, applied in the field of food extraction and detection, can solve the problems of large sample usage, time-consuming, material consumption, etc., and achieve the effects of efficient extraction and separation, improved safety, and less sample consumption.

Active Publication Date: 2019-09-17
BEIJING SANYUAN FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] 2) At present, liquid chromatography fluorescence detector is widely used to detect phospholipids, but this method cannot determine the composition of fatty acids in phospholipids
[0031] To sum up, most of the current technologies are to qualitatively and quantify phospholipids and glycerides at the molecular level respectively, and different samples need to extract lipids separately and detect them separately, resulting in a large amount of samples used, multiple pre-treatment operations, time-consuming and consuming Power and material consumption, and phospholipids are mostly quantified at the subclass level by evaporative photodetectors, that is, only the content of phosphatidylcholine can be quantified, and the content of phosphatidylcholine, etc. in different fatty acid chains cannot be determined

Method used

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  • Food lipid extraction method and detection method of lipid in food
  • Food lipid extraction method and detection method of lipid in food
  • Food lipid extraction method and detection method of lipid in food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] Example 1: Breast milk glycerides, fatty acids, phospholipids and gangliosides were extracted separately

[0124] 1 Sample pretreatment

[0125]1# tube: 200μL breast milk + 200μL ultrapure water + 1.8mL methanol + 800μL dichloromethane (chromatographically pure) → manual shaking for 10 seconds, ultrasonication for 5min → add 200μL ultrapure water + 800μL dichloromethane (chromatographically pure) → hand Shake well and oscillate for 10s→centrifuge at 6000r / min for 15min→take out the lower organic phase and transfer it to 2# tube→

[0126] 1) Upper layer of 1# tube + 1.6mL dichloromethane → centrifuge, 6000r / min, 15min → take out the lower organic phase to 3# tube

[0127] 2) The lower organic phase in 2# tube + 1mL ultrapure water + 2.2mL methanol + 0.6mL dichloromethane, shake manually → centrifuge, 3000g / min, 10min → take out the lower organic phase and mix it in 3# tube → blow dry with nitrogen → Dissolve in 1ml of dichloromethane:methanol (v / v=2:1) ​​for solid phas...

Embodiment 2

[0145] Embodiment 2 extracts breast milk lipid simultaneously

[0146] 1 Sample pretreatment

[0147] 80uL breast milk + 1200uL ultra-pure water + 2.5mL methanol + 1200uL dichloromethane (chromatographically pure) → shake manually for 10 seconds, let stand for 30min → add 1.5mL ultrapure water + 1mL dichloromethane (chromatographically pure) → shake manually Shake for 10S→centrifuge, 6000r / min, 15min→take out the lower organic phase→upper layer+1.6mL dichloromethane→centrifuge, 6000r / min, 15min→take out the lower organic phase and mix with the previous organic phase→dry with nitrogen→250uL 10mmol / Dissolve in L ammonium acetate (v / v, dichloromethane:methanol=1:1) solution → centrifuge, 6000r / min, 5min.

[0148] 2 using Kinetex C18 Column (150mm×4.6mm, 2.6μm; Phenomenex, Torrance, CA, USA) chromatographic column, column temperature 40℃, liquid phase gradient elution

[0149] The liquid phase elution gradient of table 6 embodiment 2

[0150]

[0151] 3 Mass Spectrometry ...

Embodiment 3

[0155] Example 3 Breast milk glycerides / fatty acids / phospholipids and gangliosides were extracted separately and detected twice

[0156] 1 Sample pretreatment

[0157] 100 μL of breast milk to 1# tube + 150 μL ultrapure water + 1.5 mL methanol + 800 μL dichloromethane (chromatographically pure) → manual shaking for 10 seconds, ultrasonication for 5 minutes → add 200 μL ultrapure water + 800 μL dichloromethane (chromatographically pure) → hand Shake well and oscillate for 10S→centrifuge at 6000r / min for 15min→take out the upper phase and transfer to 2# tube:

[0158] Step 1: 2# tube + 1.6mL dichloromethane → centrifuge, 6000r / min, 15min → take out the upper phase to 3# tube, wait to mix with the upper phase taken out in step 2

[0159] Step 2: The remaining organic phase in 1# tube + 2mL methanol + 0.8mL dichloromethane + 1mL ultrapure water → centrifuge, 6000r / min, 15min → take out the upper phase and mix it with the upper layer of 3# tube in step 1, the upper phase Direct f...

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Abstract

The invention provides a food lipid extraction method and a detection method of lipid in food. The food lipid extraction method comprises the following steps of: mixing and centrifuging a food sample with ultrapure water, methanol and dichloromethane for multiple times, wherein the lower layer of an organic phase obtained after multiple times of centrifugation is lipid. A small amount of samples is utilized, and extracts of different lipids through one-time treatment by using a safer lipid extraction reagent are obtained, and are subjected to qualitative and quantitative detection simultaneously or respectively. The sample consumption is reduced, the safety is improved, the consumables in the sample pretreatment and detection process are saved, the cost is reduced, and the efficiency is improved.

Description

technical field [0001] The invention relates to the field of food extraction and detection, in particular to a method for extracting food lipids and detecting food lipids. Background technique [0002] Lipids are the main nutrients in food, mainly including 8 categories: fatty acids, glycerides (including triglycerides, diglycerides, monoglycerides), glycerophospholipids (phosphatidylcholine PC, phosphatidylethanolamine PE, phospholipids Acyl inositol PI, phosphatidylserine PS, phosphatidylglycerol PG, phosphatidic acid PA), sphingolipids (ceramide Cer, sphingomyelin SM), glycolipids (gangliosides GM, GD), sterol lipids, Pregnenol ketone lipids, polyvinyls. Since the concept of lipidomics was proposed in 2003, the importance of food lipidomics to health and disease prevention has gradually been recognized. [0003] Breast milk lipids are the second largest component in human breast milk after lactose, mainly including glycerides, glycerophospholipids, sterol lipids and gly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06C11B1/10G01N33/02G01N30/88G01N33/04G01N1/4055G01N2001/4061G01N2001/4083G01N1/38
Inventor 陈历俊赵军英姜铁民乔为仓刘茜刘妍张明辉王筱迪张滢
Owner BEIJING SANYUAN FOOD
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