Al-Dirbashi Osama Y, Jacob Minnie, Al-Ahaidib Lujane Y, Al-Qahtani Khaled, Rahbeeni Zuhair, Al-Owain M, Rashed Mohamed S
Department of Genetics, King Faisal Specialist Hospital and Research Centre, P.O. Box 3354, Riyadh 11211, Saudi Arabia.
Clin Chim Acta. 2006 Mar;365(1-2):243-8. doi: 10.1016/j.cca.2005.09.001. Epub 2005 Oct 21.
Hepatorenal tyrosinemia (HT1) is considered a treatable inherited metabolic disease, particularly when detected early in life. Succinylacetone (SA), a unique metabolic marker for HT1, is normally circulating or excreted at very low physiological concentrations and is significantly increased in HT1 patients.
We developed and validated a new method for the determination of SA in urine using high-pressure liquid chromatography with fluorescence detection. SA and its homologue 5,7-dioxooctanoic acid used as internal standard (IS) were extracted from urine, derivatized with pyrenebutyric hydrazide and separated on a C18 column within 11 min. Calibration curves were linear between 0.025 to 100 micromol/l. Within- and between-day variations were <5% and results obtained by the current method compared favorably with a reference liquid chromatography tandem mass spectrometric method. The method was applied retrospectively to the analysis of urine samples from HT1 patients.
The method requires a minimal sample volume (0.1 ml) with simple instrumentation. The method enabled us to differentiate HT1 cases (n=14) from controls (n=104), regardless of the years of urine storage.
肝肾型酪氨酸血症（HT1）被认为是一种可治疗的遗传性代谢疾病，尤其是在生命早期被检测到时。琥珀酰丙酮（SA）是HT1的一种独特代谢标志物，通常以非常低的生理浓度循环或排泄，在HT1患者中显著增加。
我们开发并验证了一种使用带荧光检测的高压液相色谱法测定尿液中SA的新方法。SA及其同系物5,7-二氧辛酸用作内标（IS），从尿液中提取，用芘丁酸酰肼衍生化，并在C18柱上于11分钟内分离。校准曲线在0.025至100微摩尔/升之间呈线性。日内和日间变化均<5%，当前方法获得的结果与参考液相色谱串联质谱法相比具有优势。该方法被回顾性应用于分析HT1患者的尿液样本。
该方法所需样本量最小（0.1毫升），仪器简单。该方法使我们能够区分HT1病例（n = 14）和对照（n = 104），无论尿液储存年限如何。
