Calvin isolated the different chemicals of the Calvin cycle by chromatography.

Chromatography is a technique widely used in biochemistry and analytical chemistry to separate and identify components of a mixture. It works based on the principle of differential partitioning between a stationary phase and a mobile phase.

Calvin Cycle:
The Calvin cycle is a series of biochemical reactions that take place in the chloroplasts of plants and algae. It is the primary pathway for carbon fixation, converting carbon dioxide into glucose during photosynthesis. The cycle involves several intermediate compounds, including ribulose-1,5-bisphosphate (RuBP), glyceraldehyde-3-phosphate (G3P), and ribulose-5-phosphate (Ru5P).

Chromatography:
Chromatography is a technique used to separate mixtures into their individual components. It involves the use of a stationary phase (a solid or a liquid supported on a solid) and a mobile phase (a liquid or a gas). The mixture is applied to the stationary phase, and the individual components migrate at different rates through the stationary phase based on their chemical properties.

Isolating Chemicals of Calvin Cycle:
To isolate the different chemicals of the Calvin cycle, Calvin used chromatography. He would have likely used a technique called paper chromatography, which is a simple and inexpensive form of chromatography.

1. Preparation of Sample:
Calvin would have prepared a sample containing the chemicals of the Calvin cycle. This sample could have been extracted from plant tissue or isolated from the chloroplasts.

2. Application of Sample:
Calvin would have applied the sample onto a strip of chromatography paper. This is typically done by placing a small drop or spot of the sample near the bottom of the paper.

3. Development of Chromatogram:
The chromatography paper is then placed in a suitable solvent, which acts as the mobile phase. As the solvent moves up the paper by capillary action, it carries the individual components of the sample with it.

4. Separation of Components:
As the solvent moves up the paper, the individual components of the sample separate based on their affinity for the stationary phase and the mobile phase. Components with a higher affinity for the stationary phase will move slower, while those with a higher affinity for the mobile phase will move faster.

5. Visualization and Analysis:
Once the solvent has reached the desired height, the chromatogram is removed from the solvent and allowed to dry. The separated components appear as distinct spots or bands on the paper. These can be visualized using various techniques such as UV light, staining, or chemical reactions specific to certain components.

By comparing the chromatogram of the Calvin cycle sample with known standards or by analyzing the spots/bands, Calvin was able to identify and isolate the different chemicals of the Calvin cycle. Chromatography provided a valuable tool for studying the intermediates of the Calvin cycle and understanding the overall process of carbon fixation in photosynthesis.