FAQ

To prevent oxidation during lipid extraction, it’s essential to use an extraction method that minimizes exposure to oxygen. One effective approach is to perform the extraction in an inert atmosphere, such as nitrogen or argon, which displaces oxygen. For example, we might carry out the extraction in a glove box filled with inert gas or use sealed containers during the process. This helps ensure that sensitive lipids, such as polyunsaturated fatty acids, remain stable and intact. Additionally, incorporating antioxidants into the extraction solvent can further protect lipids from oxidative degradation. Common antioxidants include butylated hydroxytoluene (BHT) or ascorbic acid, which can scavenge free radicals and prevent lipid peroxidation. Adding BHT to a chloroform-methanol extraction solvent can help maintain the integrity of extracted lipids.

Phase separation is a crucial step in lipid extraction protocols, as it helps isolate lipids from other cellular components in a sample. Typically, a solvent mixture, such as chloroform and methanol, is used to extract lipids from biological matrices. When the mixture is added to the sample and shaken, it creates distinct phases: a lower organic phase containing the lipids and an upper aqueous phase that retains polar metabolites and other biomolecules. This separation allows for selective recovery of lipids without interference from other compounds. When extracting from tissues, we often need to optimize the volume ratios of solvents to maximize lipid recovery. Proper phase separation also minimizes the risk of co-extracting unwanted substances, such as proteins or sugars, which can complicate subsequent analyses.

Preventing lipid degradation during extraction involves implementing strategies that minimize exposure to heat, light, and oxygen. One effective method is to perform the extraction at low temperatures, such as on ice or at 4°C, to reduce enzymatic activity and chemical reactions that could lead to degradation. Using antioxidants in the extraction solvent is another critical strategy. Incorporating substances like ascorbic acid or butylated hydroxytoluene (BHT) can protect lipids from oxidation during extraction. Additionally, rapid processing is vital; samples should be extracted as soon as possible after collection to prevent any metabolic changes.

Sample was thawed on ice, whirl around 10 s, and then centrifuge it with 3000 rpm at 4 ℃ for 5 min. Take 50 uL of one sample and homogenize it with 1mL mixture (include methanol, MTBE and internal standard mixture ). Whirl the mixture for 15 min. Then add 200 uL of water and whirl the mixture for 1 min, and centrifuge it with 12,000 rpm at 4 ℃ for 10 min. Extract 500 uL supernatant and concentrate it. Dissolve powder with 200 uL reconstituted solution ,then stored in -80 ℃. Finally take the dissolving solution into the sample bottle for LC-MS/MS analysis.

We are capable of working on lymphatic fluid samples. A recommended volume of 100 μL and a minimum of 20 μL.

Yeah, we have 2 deuterated FFAs as internal standarders in our quantitative lipidomics panel.

We can perform lipid extraction and quantitative lipidomics detection for cell membrane samples. To ensure an adequate amount of membrane material for each sample, at least 10^7 cells should be used to isolate the membranes.

For cells grown in suspension: Transfer a cell suspension containing at least 2 x 10^6 cells into a 2 ml centrifuge tube. Centrifuge at 1000g for 10 minutes at 4°C, and discard the supernatant. Quickly wash the cell pellet with pre-cooled PBS (4°C) 2-3 times, each time centrifuging at 1000g for 10 minutes at 4°C and discarding the supernatant. Collect the cell pellet, snap freeze it in liquid nitrogen for 5-10 minutes, and store it at -80°C. For cells grown on dishes: Scrap the cells with small amount of media and collect at least 2 x 10^6 cells into a 2 ml centrifuge tube. Please do not use trypsin for collecting the cells. Centrifuge at 1000g for 10 minutes at 4°C, and discard the supernatant. Quickly wash the cell pellet with pre-cooled PBS (4°C) 2-3 times, each time centrifuging at 1000g for 10 minutes at 4°C and discarding the supernatant. Collect the cell pellet, snap freeze it in liquid nitrogen for 5-10 minutes, and store it at -80°C.

EPA and DHA are included in our QL database. The sample requirement of lipisomes can refer to that of EVs. For extracellular vesicle samples, the sample size required is 2*10^9 particles (NTA) recomended and 1*10^9 particles (NTA) minimum.

Yes, we have such experiences on monkey samples. The involved services include TM widely-targeted metabolomics and untargeted metabolomics, trptophan targeted and SCFA targeted metabolomics panels.

We can process this type of sample and recommend collecting sebum using Sebutape patches. We suggest using 2 patches, with at least 1 patch required for lipidomics analysis.