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建立了HPLC-紫外检测法快速测定家蚕酪氨酸羟化酶活性的方法。采用的色谱柱为Venusil XBP C18(4.6 mm×100 mm,5μm),流动相pH为3.5,流速0.5 mL/min,紫外检测波长280nm,柱温30℃。L-多巴在0.1～100μg/mL范围内线性良好,r=0.9991(n=6),L-多巴最低检测限是0.02μg/mL(S/N≥3),日内、日间相对标准偏差(RSD)在3.1%～12%之间,样品回收率在89%～104.6%之间。该方法可用于昆虫酪氨酸羟化酶(TH)活性测定。
A rapid HPLC-UV method for the determination of tyrosine hydroxylase activity in silkworms was established. The chromatographic column was Venusil XBP C18 (4.6 mm × 100 mm, 5 μm) with a mobile phase of 3.5 and a flow rate of 0.5 mL / min. The UV detection wavelength was 280 nm and the column temperature was 30 ℃. The linearity of L-dopa in the range of 0.1-100μg / mL was good, r = 0.9991 (n = 6) and the detection limit of L-dopa was 0.02μg / mL (S / N≥3) The deviation (RSD) was between 3.1% and 12% and the sample recovery was between 89% and 104.6%. This method can be used for the determination of insect tyrosine hydroxylase (TH) activity.