HPLC-MS/MS测定大鼠肝脏组织中丹参素钠的浓度

Determination of sodium Danshensu in rat liver by HPLC-MS/MS

目的建立液质联用色谱法(HPLC-MS/MS)测定SD大鼠肝脏组织中丹参素钠的浓度。方法肝脏组织样品制成匀浆后,加入内标酮洛芬,采用液液萃取方法进行预处理。色谱柱:Diamonsil C18柱(200 mm×4.6 mm,5μm);流动相:甲醇-水(含0.01‰甲酸)(80∶20);流速:1.0 ml/min;质谱条件:采用ESI离子源,负离子检测,选择MRM测定丹参素钠和酮洛芬197→135 m/z和253→209 m/z。结果丹参素钠在20.2～1 010 ng/ml检测浓度范围内呈良好的线性关系(r=0.999 8),最低定量限为20.2 ng/ml,日内精密度和日间精密度的RSD<10%,回收率在92%～107%之间。结论该方法灵敏度高、快速、简单、准确,适用于肝脏组织样品中丹参素钠的含量测定,可以满足药动学研究的需要。

Objective To establish a HPLC-MS/MS method for determination of sodium Danshensu in SD rat liver. Methods Tissue samples were pretreated by liquid-liquid extraction method after homogenate, and ketoprofen was used as the internal standard. Analytical column was D iamonsil C18 （200 mm × 4.6 mm, 5 μm）. The mobile phase was methanol： water（0.01%v formic acid） = 80 ： 20 with flow rate as 1.0 ml/min. Mass spectrum determination was performed in the MRM mode with target ions m/z 197→135 （Sodium Danshensu） , m/z 253→209 （ketoprofen） , using ESI ion resource and negative ion detection. Results The calibration curve was linear over the range of 20.2 - 1 010 ng/ml. The LLOQ of sodium Danshensu in liver was 20.2 ng/ml. The intra-and inter-day RSD were both less than 10%. The extracted recovery rate ranged from 92% to 107%. Conclusion The method was sensitive, rapid, simple and accu- rate to determine sodium Danshensu distribution in rat liver and was suitable for preclinical test of Sodium Danshensu.