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The mobile phase was composed of 40\u2010mL ethanol and 960\u2010mL buffer solution at a pH of 3.5, which was prepared by using 1\u2010hexane sodium sulfonate and anhydrous sodium sulfate, at a flow rate of 1\u2009mL\/min. The detecting wavelength was 205\u2009nm, the injection volume was 20\u2009\u03bcL, the analysis runtime was &lt;\u20097\u2009min, and the focus was on netilmicin concentration levels of 125.0\u20131000.0\u2009\u03bcg\/mL. The method's environmental friendliness was demonstrated by the conclusional AES of 87, the AGREE of 0.65, the MoGAPI of 76 with six green pieces, the BAGI score of 77.5, and the whiteness of 99.2. 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