Institute of Medical Biochemistry and Laboratory Diagnostics, Faculty General Hospital and 1st Faculty of Medicine, Charles University, Prague, Czech Republic.
HPST, s.r.o., Prague, Czech Republic.
Sci Rep. 2020 Mar 10;10(1):4411. doi: 10.1038/s41598-020-61280-z.
Although phototherapy (PT) is a standard treatment for neonatal jaundice, no validated clinical methods for determination of bilirubin phototherapy products are available. Thus, the aim of our study was to establish a such method for clinical use. To achieve this aim, a LC-MS/MS assay for simultaneous determination of Z-lumirubin (LR) and unconjugated bilirubin (UCB) was conducted. LR was purified after irradiation of UCB at 460 nm. The assay was tested on human sera from PT-treated neonates. Samples were separated on a HPLC system with a triple quadrupole mass spectrometer detector. The instrument response was linear up to 5.8 and 23.4 mg/dL for LR and UCB, respectively, with submicromolar limits of detection and validity parameters relevant for use in clinical medicine. Exposure of newborns to PT raised serum LR concentrations three-fold (p < 0.01), but the absolute concentrations were low (0.37 ± 0.16 mg/dL), despite a dramatic decrease of serum UCB concentrations (13.6 ± 2.2 vs. 10.3 ± 3.3 mg/dL, p < 0.01). A LC-MS/MS method for the simultaneous determination of LR and UCB in human serum was established and validated for clinical use. This method should help to monitor neonates on PT, as well as to improve our understanding of both the kinetics and biology of bilirubin phototherapy products.
尽管光疗（PT）是治疗新生儿黄疸的标准治疗方法，但目前尚无经过验证的用于确定胆红素光疗产品的临床方法。因此，我们的研究目的是建立一种用于临床使用的方法。为了实现这一目标，我们进行了 LC-MS/MS 测定，以同时测定 Z-胆红素（LR）和未结合胆红素（UCB）。在 460nm 光照下将 UCB 照射后，LR 被纯化。该测定方法在接受 PT 治疗的新生儿的人血清中进行了测试。样品在 HPLC 系统上用三重四极杆质谱仪检测器进行分离。该仪器的响应在 5.8 和 23.4mg/dL 之间呈线性，对于 LR 和 UCB 的检测限分别为亚微摩尔，且具有与临床医学相关的有效性参数。新生儿暴露于 PT 会使血清 LR 浓度增加三倍（p<0.01），但绝对浓度较低（0.37±0.16mg/dL），尽管血清 UCB 浓度急剧下降（13.6±2.2 与 10.3±3.3mg/dL 相比，p<0.01）。建立并验证了一种用于同时测定人血清中 LR 和 UCB 的 LC-MS/MS 方法，可用于临床。该方法应该有助于监测接受 PT 治疗的新生儿，并有助于更好地了解胆红素光疗产品的动力学和生物学。
