目标SRM对马血浆中的rhEPO-DPO进行检测和定量

简介：

Recombinant human erythropoietin (rhEPO)2 and Darbepoetin-alpha (DPO)3 are genetically engineered protein-based drugs used for the treatment of anemia by stimulating red blood cell production. The ability of these agents to stimulate red blood cell production has led to use and abuse by human and equine athletes and, thus, violates the rule of fair competition resulting in their classification as banned substances by the horse racing industry. In addition, continued administration to horses can result in anemia.3 Despite the negative aspects of rhEPO for horses, a reliable, verifiable, and legally defensible method for identification and confirmation of rhEPO/DPO has been elusive due to the very low concentrations administered. Sample collection is typically acquired only after competition, which could be in excess of 72 hours following administration. Testing of rhEPO/DPO is further confounded by the complexity of the matrices in which the drug is typically found–plasma and urine.

仪器：

Thermo Scientific TSQ Quantum Access triple quadrupole mass spectrometer

总结：

The approach presented here provides a sensitive and selective method for preparing and analyzing horse plasma for the presence of rhEPO or DPO. The advantages of this method include the ability to use up to six diagnostic peptides to confirm or refute the presence of either illegal protein-based drug.

The use of stable-isotope labeled analogues provides further means of confirming the presence of diagnostic peptides based on chromatographic retention times and ion ratios.

The sensitivity demonstrated enabled detection up to 72 hours following the last administration of rhEPO, increasing the confidence that the described method is useful in the racing industry to maintain a level field of competition.

Of particular interest is the measured sensitivity that was achieved using microspray, increasing the analysis