Zheng Joanna J, Lynch Eric D, Unger Steve E
Department of Drug Metabolism and Pharmacokinetics, The DuPont Pharmaceuticals Company, Stine-Haskell Research Center, PO Box 30, Newark, DE 19714, USA.
J Pharm Biomed Anal. 2002 Apr 15;28(2):279-85. doi: 10.1016/s0731-7085(01)00562-3.
Twenty-seven highly diversified pharmaceutical compounds were used as a test set to evaluate matrix effects from microsomal media on LC/MS analyses. The individual effects of Tris buffer, NADPH and microsomes on ESI response were investigated. Direct flow injection MS/MS analysis, using no sample preparation or HPLC separation, gave an average of 2.2-5-fold matrix suppression in MS response from Tris buffer and NADPH. More polar analytes were affected the greatest. To reduce the loss in response, an automated solid phase extraction (SPE) procedure was developed. A much smaller average matrix effect was observed when samples were prepared using a Waters Oasis HLB 96-well SPE. As little as 1 ml of methanol (MeOH) was sufficient to elute most compounds with more than 80% recovery. Comparable results were obtained by directly injecting a protein-precipitated incubation onto a fast gradient LC separation prior to MS/MS detection. No advantage was seen by using both SPE and a fast LC separation prior to MS/MS analysis.
使用27种高度多样化的药物化合物作为测试集，以评估微粒体介质对液相色谱/质谱分析的基质效应。研究了Tris缓冲液、NADPH和微粒体对电喷雾电离（ESI）响应的个体效应。采用直接流动注射串联质谱分析，不进行样品制备或高效液相色谱分离，Tris缓冲液和NADPH在质谱响应中产生了平均2.2至5倍的基质抑制。极性更强的分析物受影响最大。为减少响应损失，开发了一种自动固相萃取（SPE）方法。使用Waters Oasis HLB 96孔固相萃取柱制备样品时，观察到的平均基质效应要小得多。仅1毫升甲醇（MeOH）就足以洗脱大多数化合物，回收率超过80%。在串联质谱检测之前，将蛋白质沉淀孵育物直接进样到快速梯度液相色谱分离中，也获得了类似的结果。在串联质谱分析之前同时使用固相萃取和快速液相色谱分离没有优势。
