1仪器与实验材料-济宁医学院学报.doc

HPLC-DAD法测定洋葱中黄酮类成分的含量( * 丁玉莲 王亚楠 陈 迪 付英杰 王建安 （济宁医学院药学院，山东 日照 276826） 摘 要 目的 建立高效液相色谱（HPLC-DAD）方法，对鲜洋葱中的主要黄酮类化合物进行含量测定。方法 采用色谱柱SHIMADZU VP-ODS (250 × 4.6 mm, 5 μm)，以乙腈-水为流动相，梯度洗脱，柱温30°C，流速1.0 mL/min，检测波长350 nm。首先根据紫外光谱图，确定黄酮类化学成分。而后以芦丁为对照品建立标准曲线，对洋葱中黄酮类化学成分定量研究。结果 建立的高效液相色谱分析方法能够满足葱中主要黄酮类化学成分准确测定的要求，芦丁在5.0-502.5 μg/mL范围内有良好的线性关系（r=0.9995），重复性（RSD 4.1%），日内精密度和日间精密度（RSD 3.3 %），加样回收率（95.1%-103.2 %），检出限（0.3 μg/mL）和定量限（1.0 μg/mL）洋葱中主要黄酮类化学成分的含量为2.8 mg/g。结论 该方法简便可行，测定结果准确，能够满足洋葱中主要黄酮类化学成分准确测定的需要。 关键词：洋葱；黄酮类成分；高效液相色谱仪；含量测定 Determination of flavonoids in onion by HPLC-DAD REN Qiang DING Yu-lian WANG Ya-nan CHEN Di FU Ying-jie WANG Jian-an （Department of Pharmacy, Jining Medical College, Rizhao, Shandong 276826, People’s Republic of China） Abstract: Ojective To establish a high performance liquid chromatography coupled with a diode array detector (HPLC-DAD) method for the determination of major flavonoids in onion. Methods The chromatographic separation was performed on a SHIMADZU VP-ODS (250 × 4.6 mm, 5 μm) column. The mobile phase was composed of water (A) and acetonitrile (B) using gradient elution. The column temperature was 30°C, the flow rate 1.0 mL/min, and UV absorbance was detected at a wavelength of 350 nm. Firstly, the flavonoids were identified by maximum UV absorption wavelength. Then,?the rutin used as a standard substance?to establish the standard curve to determine the major?flavonoids?in onion. Results The HPLC-DAD method was established and validated to determine the major flavonoids. The calibration curve was linear in the range of 5.0-502.5 μg/mL with good linearity (r=0.9995), repeatability (RSD 4.1 %), intra-day and inter-day precision (RSD 3.3%), recovery (95.1%-103.2 %), limit of detection (0.3 μg/mL), and limit of quantification (1.0 μg/mL). The content of?flavonoids?in onion?was 2.8 mg/g. Conclusion The method is simple, feasible, and accurate approach for?the determination major?flavonoids in?onion. Key words: Onion; Flavonoids; HPLC-