Grebe Stefan Kg, Singh Ravinder J
Departments of Laboratory Medicine & Pathology and Medicine.
Clin Biochem Rev. 2011 Feb;32(1):5-31.
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has seen enormous growth in clinical laboratories during the last 10-15 years. It offers analytical specificity superior to that of immunoassays or conventional high performance/pressure liquid chromatography (HPLC) for low molecular weight analytes and has higher throughput than gas chromatography-mass spectrometry (GC-MS). Drug/Toxicology and Biochemical Genetics/Newborn Screening laboratories were at the vanguard of clinical LC-MS/MS use, but have been eclipsed by Endocrine laboratories. In USA reference/referral laboratories, most steroids and biogenic amines are now assayed by LC-MS/MS, and the technology has started to penetrate into smaller laboratories. Assays for mineralo- and gluco-corticoids and their precursors, sex steroids, metanephrines and 25-hydroxy vitamin D highlight the advantages of LC-MS/MS.However, several limitations of LC-MS/MS have become apparent, centring on the interacting triangle of sensitivity - specificity - throughput. While sample throughput is higher than for conventional HPLC or GC-MS, it lags behind automated immunoassays. Techniques which improve throughput include direct sample injection, LC-multiplexing and samplemultiplexing. Measures to improve specificity and sensitivity include sample clean-up and optimising chromatography to avoid interferences and ion suppression due to sample-matrix components. Next generation instrumentation may offer additional benefits.The next challenge for clinical LC-MS/MS is peptide/protein analysis. The quest for multi-biomarker profiles for various diseases has largely failed, but targeted peptide and protein testing by LC-MS/MS, directed at analytical and clinical questions that need to be answered, is proving highly successful. We anticipate that this will result in similar growth of clinical protein/peptide LC-MS/MS as has been seen for low molecular weight applications.
在过去10到15年里，液相色谱-串联质谱法（LC-MS/MS）在临床实验室中得到了巨大发展。对于低分子量分析物，它提供了优于免疫测定法或传统高效/高压液相色谱法（HPLC）的分析特异性，并且比气相色谱-质谱法（GC-MS）具有更高的通量。药物/毒理学和生化遗传学/新生儿筛查实验室是临床LC-MS/MS应用的先锋，但已被内分泌实验室超越。在美国的参考/转诊实验室中，现在大多数类固醇和生物胺都通过LC-MS/MS进行检测，并且该技术已开始渗透到较小的实验室。对盐皮质激素、糖皮质激素及其前体、性激素、甲氧基肾上腺素和25-羟基维生素D的检测突出了LC-MS/MS的优势。然而，LC-MS/MS的几个局限性已经显现出来，主要集中在灵敏度-特异性-通量的相互作用三角关系上。虽然样品通量高于传统HPLC或GC-MS，但仍落后于自动化免疫测定法。提高通量的技术包括直接进样、液相色谱多路复用和样品多路复用。提高特异性和灵敏度的措施包括样品净化和优化色谱以避免由于样品基质成分导致的干扰和离子抑制。下一代仪器可能会带来更多益处。临床LC-MS/MS的下一个挑战是肽/蛋白质分析。寻找各种疾病的多生物标志物谱在很大程度上失败了，但通过LC-MS/MS针对需要回答的分析和临床问题进行靶向肽和蛋白质检测已被证明非常成功。我们预计这将导致临床蛋白质/肽LC-MS/MS出现与低分子量应用类似的增长。
