Reliable prenatal diagnosis of Canavan disease by measuring N-acetylaspartate in amniotic fluid using liquid chromatography tandem mass spectrometry

• Published in: Prenatal diagnosis 2009-05, Vol.29 (5), p.477-480
• Main Authors: Al-Dirbashi, Osama Y., Kurdi, Wesam, Imtiaz, Faiqa, Ahmad, Asmahan M., Al-Sayed, Moeenaldeen, Tulbah, Maha, Al-Nemer, Maha, Rashed, Mohamed S.
• Format: Article
• Language: English
• Subjects: Amidohydrolases - genetics; amniotic fluid; Amniotic Fluid - chemistry; Aspartic Acid - analogs & derivatives; Aspartic Acid - analysis; Biological and medical sciences; canavan disease; Canavan Disease - diagnosis; Canavan Disease - etiology; Canavan Disease - genetics; Canavan Disease - pathology; Case-Control Studies; Chromatography, Liquid - methods; Delivery. Postpartum. Lactation; Female; Fundamental and applied biological sciences. Psychology; Genetics of eukaryotes. Biological and molecular evolution; Gestational Age; Gynecology. Andrology. Obstetrics; Humans; liquid chromatography tandem mass spectrometry; Medical sciences; Molecular and cellular biology; Mutation - physiology; N-acetylaspartic acid; Pregnancy; prenatal diagnosis; Prenatal Diagnosis - methods; Reproducibility of Results; Risk Factors; Tandem Mass Spectrometry - methods
• ISSN: 0197-3851; 1097-0223
• DOI: 10.1002/pd.2223

Objective Prenatal diagnosis of Canavan disease by measuring N‐acetylaspartic acid (NAA) in amniotic fluid is reliable and preferred over aspartoacylase enzyme assay especially in populations with unknown mutations. Typically based on GC–MS, existing methods are time‐consuming and laborious. We developed a novel LC–MS/MS method for determination of NAA in amniotic fluid with minimal sample preparation. Method NAA and d3‐NAA were detected by negative‐ion electrospray ionization‐MS/MS. Quantification was achieved by standard addition using six 0.1 mL portions of each specimen enriched with increasing NAA amounts (0, 0.05, 0.1, 0.2, 0.3, and 0.4 µg) and endogenous NAA was calculated by extrapolation. Results Injection‐to‐injection time was 2 min whereas the turn around time from sample receipt was about 1 h. Intraday (n = 10) and interday (n = 10) variations were less than 9.4%. The reference range determined using gestation‐matched controls (n = 12) of 1.1–2.7 µmol/L is in agreement with the literature. Specimens from at‐risk pregnancies with established diagnosis (n = 4) were successfully analyzed. Conclusion We developed a new method that enables reliable, sensitive, and selective determination of NAA in a small volume of amniotic fluid for the prenatal diagnosis of Canavan disease. The simple sample preparation adopted in this work precluded the necessity for extraction and derivatization. Copyright © 2009 John Wiley & Sons, Ltd.