超高效液相色谱法检测药桑椹中1-脱氧野尻霉素

建立超高效液相色谱测定新疆药桑椹中1-脱氧野尻霉素（1-deoxynojirimycin，DNJ）含量的方法。药桑椹粉经0.05 mol/L盐酸提取，在pH 8.5的硼酸盐缓冲液条件下，氯甲酸-9-芴基甲酯（9-fluorenylmethyl chloroformate，FMOC-Cl）与DNJ反应生成具有紫外吸收的络合物（DNJ-FMOC），然后采用超高效液相色谱-紫外检测器测定。液相色谱条件为CORTECSTM C18色谱柱（2.1 mm×100 mm，2.7 μm），流动相为乙腈-0.5%醋酸（28∶72，V/V），在流速0.6 mL/min、柱温28 ℃、检测波254 nm条件下检测。结果表明：DNJ与其他组分分离效果良好，方法检出限为0.1 μg/mL（RSN=3），线性范围在0.5～20 μg/mL，相关系数（r）为0.999 970，加标回收率平均为106.4%。精密度实验、稳定性实验、重复性实验以及回收率实验表明，该方法稳定可靠，可作为测定药桑椹中DNJ含量的有效方法。

Determination of 1-Deoxynojirimycin in Black Mulberry Fruit by Ultra Performance Liquid Chromatography

The contents of 1-deoxynojirimycin (DNJ) in the fruits of black mulberry (Morus nigra L.) were measured using
ultra performance liquid chromatography (UP LC). Dried fruits were pulverized and extracted with 0.05 mol/L HCl, and then
incubated with 9-fluorenylmethyl chloroformate (FMOC-Cl) in pH 8.5 borate buffer to produce the complex DNJ-FMOC.
The concentration of DNJ-FMOC was detected by UP LC coupled with UV detector. A CORTECSTM C18 column was used
and the column temperature was kept at 28 ℃. The mobile phase was acetonitrile-0.5% acetic acid solution (28:72, V/V) with
a flow rate of 0.6 mL/min. The UV detection wavelength was 254 nm. The results indicated that DNJ was separated well
with a linearity ranging from 0.5 to 20 μg/mL. The limit of detection (LOD) was 0.1 μg/mL and the correlation coefficient (r)
between the peak area of DNJ-FMOC and DNJ concentration was 0.999 970 and the average recovery from spiked samples
was 106.4%. The tests for precision, stability, reproducibility and recovery rate showed that the method was stable and
reliable, and could be used as an effective method for the determination of DNJ content in black mulberry fruits.