Chromatography used for testing Swine Flu

UPDATE: Article from TheScientist.com – Can biotech tackle swine flu? – click here to read

For decades, scientists have been using Thin Layer Chromatography, HPLC, gas, and other forms of chromatography to study swine influenza.

Scientists working on swine fluHere is just one example. In October of 1985, a team from the Department of Neurology at The Medical University of South Carolina published “Lipid content of swine influenza and other vaccines” – here’s a couple of excerpts:

ABSTRACT: An analysis of the lipids in swine influenza vaccines was performed, comparing six different lots of swine influenza, other influenza and noninfluenza vaccines.
Cholesterol content and phospholipid content varied greatly, but there were no major differences between the types of vaccines. Appreciable amounts of phosphatidylethanolamine were found in only one swine influenza vaccine. The major phospholipids of influenza vaccines were phosphatidylcholine, sphingomyelin and phosphatidic acid. A detectable amount of phosphatidylserine was not found in any swine influenza vaccine, but was present in two of three nonswine influenza vaccines. Only two of six swine influenza vaccines showed trace amounts of ganglioside. However, larger quantities of galactocerebroside were found in all
influenza vaccines examined, including swine influenza vaccines.

Neutral lipids were separated on silica gel thin layer chromatography (TLC) plates developed in light petroleum ether/diethyl ether (96:4, v/v) and visualized by exposure to iodine vapors.
Phospholipids were separated by two-dimensional TLC using high performance TLC (HPTLC) plates. After application, samples were chromatographed in C/M/concentrated ammonia (65:35:5, v/v/v) to the top of the plate plus an additional 10 min. HPTLC plates were air-dried and held in vacuo overnight over P205 to reactivate the silica gel. Chromatography in the second direction was performed in chloroform/acetone/methanol/acetic acid/water (5:2:1:1:0.5, v/v/v/v/v). After being air-dried, phospholipids were visualized by exposure to iodine vapors, matched to standards and marked. After sublimation of I2, marked areas were carefully scraped from the glass backing, charred and assayed for liberated phosphate by the method of Ames {27}. Prior to TLC, aliquots were withdrawn and assayed in the same manner for total phospholipid determination.

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