目的：建立测定大鼠血浆及粪便中新疆紫草3种萘醌类成分含量的高效液相色谱法（HPLC 法）。方法采用 COSMOIL C18-AR-Ⅱ（4．6 mm×250 mm，5μm）色谱柱，流动相 A 乙腈（A）-0．15％甲酸（B）（v／v）溶液，进行洗脱，流速1．0 mL／min，检测波长275 nm ，柱温25℃，检测时间35 min。结果血浆中左旋紫草素在0．40～6．40μg、乙酰紫草素在25．0～600．0μg、β，β′-二甲基丙烯酰紫草素在25．0～600．0μg 范围内均与峰面积呈线性关系，左旋紫草素回收率为97．78％～100．1％、乙酰紫草素为100．1％～100．2％、β-β′-二甲基丙烯酰紫草素为97．78％～100．1％，日间、日内 RSD 均＜1．5％。粪便中各组分在10～400μg／mL 范围内与峰面积呈线性关系，平均回收率分别为（78．2±2．5）％、（82．0±2．9）％和（79．5±3．5）％。0～12、12～24、24～48 h 粪便中左旋紫草素绝对回收量分别为23．61、38．28、4．13 mg；乙酰紫草素分别为7．20、70．78、11．53 mg；β-β′二甲基丙烯酰紫草素分别为46．32、60．15、16．88 mg。结论HPLC 法准确可靠，重复性好，适用于测定血浆及粪便中新疆紫草萘醌类成分含量。%Objective To establish a HPLC method for determine of three contents of naphthaquinone from Arnebia euchroma (Royle)Johnst in rats plasma and faeces.Methods COSMOIL C18-AR-Ⅱ (4.6 mm× 250 mm,5 μm),column was used with acetonitrile (A)-0.15% formic acid (B)(v/v)as mobile phase by gradient eluting with the flow rate of 1.0 mL/min for 35 min,and the column temperature was maintained at 25℃,detecting wavelength at 274 nm.Results The linear range in plasma of shikonin,acetylshikonin andβ-β,-dimethylacrylshikonin were between 0.40 to 6.40 μg,25.0 to 600.0 μg and 25.0 to 600.0 μg,re-spectively.The recoveries of these ingredients correspondingly were between 97.78 to 100.1%,100.1 to100.2% and 97.78 to100.1%.The RSD of intra-day and inter-day were less than 1.5%.The linear range in faeces of shikonin,acetylshikonin andβ-β,-dimethylacrylshikonin were between 10-400 μg/mL with the average recoveries of (78.2 ± 2.5)%,(82.0 ± 2.9)% and (79.5 ± 3.5)%,respectively.Absolute recovery amount of Naphthoquinone in feces as:shikonin 0-12 h:23.61 mg,12-24 h:38.28 mg,24-48 h:4.13 mg;acetylshikonin 0-12 h:7.20 mg,12-24 h:70.78 mg,24-48 h:11.53 mg andβ-β,-dimeth-ylacrylshikonin 0-12 h:46.32 mg,12-24 h:60.15 mg,24-48h:16.88 mg.Conclusion The established method has a good selectivity and reproducibility for the determination of naphthaquinone from Arnebia euchroma (Royle)Johnst in plasma and faeces of rats.
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