Sphingolipid Profiling LC-MS/MS
The facility has a Shimadzu Prominence UPLC system equipped with a CBM-20A controller, a DGU-A3 degasser, three UPLC solvent delivery modules LC-ADXR, a CTO-20AC column oven/chiller, a SIL-20ACTHT autosampler. The UPLC system is attached to an API 5000 LC-MS/MS system (Applied Biosystems/MDS SCIEX, Concord ON, Canada).
The API 5000TM LC-MS/MS system is a very high sensitivity triple-quadrupole mass spectrometer for small-molecule analysis.
Dynamic Range: 5 orders of magnitude
Ionization Sources: Turbo V TM implemented with TurboIonSpray® and APCI probes.
Mass Range: 5-1,250 Da
Scan Types: Q1 MS, Q3 MS, Product Ion, Precursor Ion, Neutral Loss or Gain, MRM
The analytical method is applicable to biological samples of all types (plasma, serum, urine, cell culture and culture media, tissue, etc.).
Guidelines for All Sample Types
Consistency in sample handling and collection is critical. For samples belonging to the same study, please avoid changes in the sample collection technique, time of sampling, time from collection to freezer, etc.
Samples should be frozen immediately (or as soon as possible) after collection and should be stored at -80°C until shipping.
For plasma, serum, and pulverized tissue samples, please use polypropylene cryovials (for volumes of plasma and serum smaller than 100 microliters, use polypropylene microcentifuge tubes).
For adipose tissue samples and non-pulverized tissue samples, please use culture, screw thread, borosilicate glass tubes 13x100 mm (Fisher Scientific Cat # 14-962A) with phenolic screw caps (Fisher Scientific Cat # 14-962A)
Required Sample Amounts
|Sample Type||Sample Size|
|Human plasma/serum||200 microliters|
|Mouse plasma/serum||50-75 microliters|
|Tissue samples||30-40 mg|
Please label sample tubes with permanent markers while tubes are warm (cold tubes will not hold the ink) or use specialized freezer-safe labels. Include as much information as possible: sample volume, weight, etc. Please exclude any patient identifiers from human samples.
Please label samples with a series of sequential numbers.
Samples should be frozen as soon as possible after collection and stored at -80°C until shipping. Flash freezing in liquid nitrogen is ideal. However, if these resources are not available, immediately placing tubes in a -80°C freezer is acceptable.
Samples should be shipped on dry ice including a hard copy of the list of samples contained in the shipment.
|Batch Size||Cost for UT Southwestern Investigators|