Adenosine triphosphate (ATP) is considered to be a universal energy source that is essential for cell
synthesis in the survival and reproduction of all organisms. ATP can be produced through a variety of
cellular pathways. The most typical example is synthesis by adenosine triphosphate synthase through
oxidative phosphorylation in mitochondria, or synthesis by photosynthesis in plant chloroplasts. The main
energy sources for ATP synthesis are glucose and fatty acids.
ATP has an absorption peak at 254 nm, and its content can be determined by high performance liquid
chromatography.
Reagents and Equipment Required but Not Provided：
High-efficiency liquid chromatograph (C18 column (4.6×250 mm), ultraviolet detector (VWD)),
desktop centrifuge, adjustable pipette, mortar/ homogenizer, brown EP tube, 50 syringe filters (water, 0.45
µm), syringe, suction filter, filter membrane (organic, water), 50 brown injection bottle (2 mL), acetonitrile
(chromatographically pure, 500 mL), ultrapure water.
Preparations before the experiment:
1. Use 500 mL of chromatographically pure acetonitrile (mobile phase A) and 1000 mL of configured
mobile phase B to filter with a filter membrane to remove impurities in the solvent to prevent clogging the
chromatographic column. (Acetonitrile uses 0.45 µm organic filter membrane for suction filtration, and the
configured mobile phase B uses 0.22 µm aqueous filter membrane for suction filtration).
2. Ultrasound the prepared mobile phases A and B for 30 minutes to remove the gas in the solvent to
prevent clogging the chromatographic column and affecting the experimental results.
3. Preparation of standard: 1 μmol/mL ATP-Na2 standard solution is diluted with distilled water to 0.5
μmol/mL, 0.1 μmol/mL, 0.05 μmol/mL, 0.01 μmol/mL, 0.005 μmol/mL ATP standard solution. (The
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