MacNeill Robert, Hutchinson Tisha, Acharya Vishva, Stromeyer Ryan, Ohorodnik Susan
At time of publication: Covance - Bioanalysis, PO Box 2360 Mettlers Road, East Millstone, NJ 08875-2360, USANote: At time of writing: Envigo CRS, PO Box 2360 Mettlers Road, East Millstone, NJ 08875-2360, USA.
Bioanalysis. 2019 Jun;11(12):1157-1169. doi: 10.4155/bio-2019-0031. Epub 2019 Jun 26.
Reliable quantitative LC-MS methodology has been established and validated for an oligonucleotide in plasma in a fresh and unique fashion, free of ion-pairing reagents and the various associated deleterious effects from primary solution preparation through sample preparation and extraction to the LC-MS analytical end point, offering a highly selective mixed-mode solid-phase extraction with hydrophilic-interaction liquid chromatography as the chromatographic element prior to SRM detection. Inter- and intra-assay accuracy and precision ranged from 97.9 to 111% and 2.75 to 9.66%, respectively. Recoveries of 50% were attained, and there was no significant matrix effect manifestation. The method demonstrated rugged performance and reliability under the optimized conditions, indicating a possible exciting new avenue, free of ion-pairing, for general application in oligonucleotide quantitative LC-MS.
已经以新颖独特的方式建立并验证了一种可靠的定量液相色谱 - 质谱方法，用于分析血浆中的寡核苷酸。该方法无需离子对试剂，且从原始溶液制备到样品制备、萃取再到液相色谱 - 质谱分析终点，不存在各种相关的有害影响。在进行SRM检测之前，提供了一种高度选择性的混合模式固相萃取方法，并以亲水相互作用液相色谱作为色谱分析元件。批间和批内的准确度和精密度分别在97.9%至111%和2.75%至9.66%之间。回收率达到50%，且未表现出明显的基质效应。该方法在优化条件下表现出良好的耐用性和可靠性，表明这是一条可能令人兴奋的、无需离子对的新途径，可广泛应用于寡核苷酸定量液相色谱 - 质谱分析。
