Huang Jianming, Li Ni, Yu Yunqiu, Weng Weiyu, Huang Xubing
Department of Pharmacognosy, School of Pharmacy, Fudan University, Shanghai 200032, PR China.
J Pharm Biomed Anal. 2006 Feb 13;40(2):465-71. doi: 10.1016/j.jpba.2005.07.051. Epub 2005 Sep 21.
A specific, precise and accurate high-performance liquid chromatography (HPLC) method for the determination of aglycone conjugated metabolites of scutellarin in plasma after enzymolysis to scutellarein (the aglycone of scutellarin) was developed and validated. The chromatographic separation was performed on a Lunar C18(2) reversed-phase column at a column temperature of 40 degrees C. The mobile phase, delivered at 1.0 ml/min, consisted of acetonitrile-KH2PO4 buffer (40 mM, pH 2.5) (33:67, v/v). The detection wavelength was set at 335 nm. Scutellarein and I.S. (quercetin) were isolated by a liquid-liquid extraction after incubating the plasma samples with beta-glucuronidase/sulfatase. The method was validated using scutellarin spiked plasma as standards. Linearity was confirmed in the concentration range of 0.2165-4.329 nmol/ml, R.S.D.s were within 8.32%, and the recoveries of scutellarein ranged from 101.2 to 108.6%. The method is applicable to the pharmacokinetic study of aglycone conjugated metabolites of scutellarin in rats after oral administration of scutellarin.
建立并验证了一种特异性强、精密度高且准确的高效液相色谱（HPLC）方法，用于测定血浆中灯盏花素经酶解生成的灯盏花素苷元（灯盏花素的苷元）的共轭代谢产物。色谱分离在Lunar C18(2)反相柱上进行，柱温为40℃。流动相以1.0 ml/min的流速输送，由乙腈 - KH2PO4缓冲液（40 mM，pH 2.5）（33:67，v/v）组成。检测波长设定为335 nm。在血浆样品与β-葡萄糖醛酸酶/硫酸酯酶孵育后，通过液 - 液萃取分离灯盏花素苷元和内标物（槲皮素）。该方法以加标灯盏花素血浆作为标准进行验证。在0.2165 - 4.329 nmol/ml的浓度范围内确认了线性关系，相对标准偏差（R.S.D.）在8.32%以内，灯盏花素苷元的回收率在101.2%至108.6%之间。该方法适用于大鼠口服灯盏花素后灯盏花素苷元共轭代谢产物的药代动力学研究。
