摘要
目的 建立测定大鼠血清和肝、肾、小肠组织中芦丁和槲皮素的高效液相色谱法。方法 采用ODS-SPC18色谱柱（4.6 mm×250 mm,粒径5μm）,以乙腈（B）和0.5%磷酸缓冲液（A）为流动相梯度洗脱（25%B 0-6min,25%-50%B6-10min,50%B 10-17min,50%-25%B 17-20min,25%B 20-25 min）;流速1ml/min;柱温40℃,检测波长360 nm,测定大鼠血清和组织中芦丁和槲皮素含量。结果 血清芦丁在0.08-25 mg/ml,槲皮素在0.016-10 mg/ml范围内线性关系良好,相关系数为0.9999;芦丁和槲皮素的检测限分别为0.016μg/ml和0.004mg/ml;日内RSD分别小于7.4%和8.1%,日间RSD小于9.1%和9.0%。肝、肾、小肠组织中芦丁和槲皮素在0.05-8mg/ml范围内线性关系良好,相关系数为0.9999,肝组织芦丁和槲皮素的检测限分别为0.012 mg/ml和0.004 mg/ml,日内RSD分别小于7.9%和8.9%,日间RSD分别小于8.9%和7.2%,肾脏组织和小肠的芦丁和槲皮素检测限均为0.02mg/ml;血清和肝脏、肾脏、小肠组织中芦丁和槲皮素的回收率均在72%以上。结论 该方法灵敏度较高、重复性良好,操作简单、快速,可用于血清和组织中芦丁和槲皮素的分析测定。
Objective To establish a HPLC method to determine rutin and quercetin in rat serum and liver, kidney, small intestine. Methods The ODS-SPC18 chromatography column （4.6 mm × 250 mm, 5μm） was used and acetonitrile （B） and 0.5% phosphate buffer （A） as the mobile phase with gradient elution （25% B 0-6min, 25%-50%B 6- 10 min, 50%B 10-17 min, 50%-25%B 17-20min, 25%B 20-25min）. Flow rate was 1ml/min and column temperature 40℃. Detection wavelength was 360 nm. Results The method had good linearity in the concentration ranges of 0.08- 25μg/ml and 0.016-10 μg/ml for serum rutin and quereetin, respeceively. The correlation coefficient （r） was 0.9999. The detection limits of rutin and quereetin were 0.016 μg/ml and 0.004 μg/ml, respedetively. Interday precision （RSD,%） was less than 7.4% and 8. 1%. Intraday precision （RSD,%） less than 9.1% and 9.0%. The rutin and quercetin in liver, kidney, small intestine had good linearity in the concentration ranges of 0.05 -8 μg/ml. The correlation coefficient （r） was 0.9999. The detection limits of rutin and quereetin in liver were 0.012 μg/ml and 0.004 μg/ml, respectively. Interday precision （RSD,%） was less than 7.9% and 8.9% and intraday precision （RSD,%） less than 8.9% and 7.2%. The detection limits of rutin and quercetin in kidney and small intestine were all 0.020 μg/ml. The recoveries of rutin and quereetin in serum and liver were more than 72% Conelusion The established method is sensitive and reproducible and can be used in determination of rutin and quereetin in serum and tissues.
出处
《营养学报》
CAS
CSCD
北大核心
2014年第4期383-387,共5页
Acta Nutrimenta Sinica
