Benson: Instantly.   Separation of 14C-products Buchanan: Instantly. And then how did you identify

the products that had been formed?   Benson: Well, you separate them by filter paper chromatography.   Buchanan: How did you use paper chromatography to separate the products? Could you describe that? Here’s a paper chromatogram. What did you do to separate the compounds?   Benson: Well, you put all the products at the origin—let’s IWP-2 mouse say the origin is here—and then develop it in this direction first, by putting it in a trough—dipped in phenol saturated with water. And it goes through the paper. And then you turn it—   Buchanan: One of the solvents used in the second dimension was butanol propionic acid water. Did you develop that solvent?   Benson: Oh, yeah.   Buchanan: Yes. So the combination of phenol water and butanol propionic acid water turned out to be very effective. And it was used subsequently by laboratories around the world.   Benson: Fortunately, I did an experiment with the compounds moving in the paper. And, of course, the paper absorbs the water but not the other organic compounds. So as it moves, the solvent characteristics kept changing. So that greatly enhanced the function of the second solvent.

  Buchanan: Who advised you to use two-dimensional paper chromatography?   Benson: Oh, it was invented in England. But they had stupid solvents that were absolutely poisonous. And the physicists were upstairs, who were—using a drier for the paper chromatograms. They—they were getting sick. And that just means a change of solvents, so they could tolerate them better.   Buchanan: So the originators of the technique were Martin and Synge?   Benson: Yeah.   Buchanan: And at Berkeley, Astemizole you were in the same building with the physicists.   Benson: Yeah.   Buchanan: Was this the old Radiation Laboratory?   Benson: Yeah.   Benson: It was all physicists. When—when we moved in, they had uranium all over

the floor, which was a little bit radioactive. So I—I got some cheap linoleum and placed it on top of it. And that blocked it off. And we—   Benson: —we didn’t have any chemical hoods in the laboratory, where you could work with things and the air would be exhausted out the top. We just had big windows. And we opened the Entinostat windows and hoped for the best. And all the amino acids, like alanine, glutamic acid, they traveled different distances.   Buchanan: And so the 3-phosphoglycerate was separated from—   Benson: It goes—   Buchanan: —the sugar phosphates   Benson: —would go up here.   Buchanan: So you probably learned to recognize that as a very bright spot—   Benson: Yeah.   Buchanan: —in short-exposure—   Benson: Very dark spot.   Buchanan: —samples. And then how did you locate the compounds that were labeled in the photosynthesis experiments?   Benson: We did—by Geiger counters, just scan them.   Buchanan: So you got the major ones that way. But the minor ones, you had to go to the technique of radioautography.   Benson: Well, yeah.

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