Table 1.
Estimated amounts for sample preparation
| Sample | Amount for extraction | Injection volume (μL) | Actual sampling amount | Peak intensitya | Identified number of phospholipids species |
|---|---|---|---|---|---|
| Yeast cells | 1–4 OD600 units | 5 | <0.01 OD unit | 105–106 | ∼80 |
| Mammalian cells (e.g., HEK293T) | ≥100,000 cells | 5 | >20 cells | 105–106 | ∼300 |
| Mouse serum | 5 μL | 5 | ∼0.1 μL | 105–106 | ∼100 |
| Tissue (e.g., kidney) | 5 mg | 5 | ∼10 μg | 105–106 | ∼200 |
Note: The actual sampling amount is calculated according to a dilution rate used in this protocol. We usually take a 200 μL aliquot from 2.8 mL of the chloroform fraction of the extraction solution. The lipid extract is dried and re-dissolved in a 200 μL sampling buffer. 5 μL is injected for analysis.
a
Peak intensity designates an estimated range for phospholipid peaks with high signals.