Anoectochilus roxburghii was used as the test material, and the white light with an optical density of 30 μmol·m–2·s–1 was supplemented with three intensities of 60 μmol·m–2·s–1, 120 μmol·m–2·s–1 and 180 μmol·m–2·s–1 blue light stress for 56 days. Aminoquinoline-N-hydroxysuccinyl carbamate (AQC) pre-column derivatization combined with ultra performance liquid chromatography-tandem mass spectrometry was used to determine the content of free amino acids in order to clarify the response characteristics of the free amino acid components for blue stress. A total of 22 amino acids were detected stably. Threonine, Alanine, Glutamine, Valine, Sarcosine, Histidine, Asparagine and Tyrosine were the characteristic amino acids in A. roxburghii. The total amount of free amino acids increased first and then decreased with the extension of the stress time, and the content was the highest on the 14th day. The comprehensive results showed that blue light stress for 14 days could promote the synthesis of a variety of free amino acids in A. roxburghii, and the effect was best when supplemented with 180 μmol·m–2·s–1 intensity of blue light.
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