A preparation method for a near-infrared fluorescent tracer used for diagnosing a metastatic lymph node, as follows: 6-aminocaproic acid (Fmoc-acp-OH) is used with cRGDyK to produce Fmoc-acp-cRGDyK; when deprotected, the amino group produces Fmoc-glu-acp-{cRGDyK}2 with a glutamate, then reacts with Fmoc-acp-OH to produce Fmoc-acp-glu-acp-{cRGDyK}2; the Fmoc group is deprotected to produce NH2-acp-glu-acp-{cRGDyK}2, and then dehydrated and condensed with ZW800-COOH to produce ZW800-glu-acp-{cRGDy}2; and separation and purification are performed. A ZW800-bi-cRGDyK product prepared as such has a high yield, effectively reduces costs of similar products, and is easy to industrialize and promote.
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