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.
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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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