Al-Dirbashi Osama Y, Rashed Mohamed S, Jacob Minnie, Al-Ahaideb Lujane Y, Al-Amoudi Mohamed, Rahbeeni Zuhair, Al-Sayed Moeen M, Al-Hassnan Zuhair, Al-Owain Mohamed, Al-Zeidan Hamad
National Laboratory for Newborn Screening, Department of Genetics, King Faisal Specialist Hospital and Research Centre, PO Box 3354, Riyadh 11211, Saudi Arabia.
Biomed Chromatogr. 2008 Nov;22(11):1181-5. doi: 10.1002/bmc.1049.
We describe an improved diagnostic method for tyrosinemia type 1 based on quantifying succinylacetone in dried blood spots by ultra-performance liquid chromatography tandem mass spectrometry. Succinylacetone extracted from a single 3/16 inch disk of specimen collection paper containing a dried blood spot was derivatized with dansylhydrazine, separated on an Acquity UPLC BEH C(18) column (2.1 x 50 mm, 1.7 microm) and detected by electrospray ionization tandem mass spectrometry. Succinylacetone derivative eluted at 0.6 min with a complete run time of 1 min. Using a 13C4 labeled succinylacetone as an internal standard, the calibration plot was linear up to 100 micromol/L with a detection limit (S/N = 3) of 0.2 micromol/L. Intra-day (n = 13) and inter-day (n = 10) variations were better than 10%. The cutoff level of succinylacetone in dried blood spots from healthy infants obtained by the current method was 0.63 micromol/L (n = 151). In dried blood spots from patients with established tyrosinemia type 1 (n = 11), concentration of succinylacetone was 6.4-30.8 micromol/L.
我们描述了一种基于超高效液相色谱串联质谱法定量干血斑中琥珀酰丙酮的1型酪氨酸血症改进诊断方法。从含有干血斑的单个3/16英寸标本采集纸圆片中提取的琥珀酰丙酮用丹磺酰肼衍生化，在Acquity UPLC BEH C(18)柱（2.1×50 mm，1.7微米）上分离，并通过电喷雾电离串联质谱法检测。琥珀酰丙酮衍生物在0.6分钟洗脱，整个运行时间为1分钟。使用13C4标记的琥珀酰丙酮作为内标，校准曲线在高达100微摩尔/升时呈线性，检测限（信噪比=3）为0.2微摩尔/升。日内（n = 13）和日间（n = 10）变异均优于10%。通过当前方法获得的健康婴儿干血斑中琥珀酰丙酮的临界水平为0.63微摩尔/升（n = 151）。在确诊为1型酪氨酸血症的患者（n = 11）的干血斑中，琥珀酰丙酮浓度为6.4 - 30.8微摩尔/升。
