Development of an analytical method for determination of urine 3-phenylpyruvic acid based on liquid-liquid extraction and LC-MS/MS
Küçük Resim Yok
Tarih
2024
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Aim: The aim of this study is to develop a rapid, precise, specific LC-MS/MS method for the determination of 3-phenylpyruvic acid, which has an important place in the diagnosis of phenylketonuria, a metabolic disease resulting from Phenylalanine Hydroxylase Enzyme deficiency. Materials and Methods: Analytical measurements were made with Acquity UPLC MS MS (Waters Xevo TQD). The chromatographic separation was operated on an Acquity UPLC Phenyl column (50 mm × 2.1 mm, 1.7 ?m) with gradient elution using 0.1% formic acid containing water and methanol as the mobile phase. Within the scope of the study, firstly, sample preparation steps were focused, and dispersive liquid-liquid extraction and traditional liquid extraction methods were examined. The best results were obtained in the conventional liquid-liquid extraction method, in which dichloromethane was used as the extraction solvent. Furthermore, trans-cinnamic acid molecule was used as an internal standard for the determination of 3-Phenylpyruvic acid in this study and tested according to the validation steps. Results: The linear range of the developed LC MS MS method was found to be between 0.009-5 ?M, while the detection limit was found to be 0.001 ?M. Intraday repeatability was below 7% for 3 levels, and interlay repeatability was below 10%. In the recovery trial, which showed the accuracy of the method, the results in the urine sample for 3 levels were in the range of 97%-103%, and there was no significant ion suppression in the matrix effect trial, which showed how clean the final matrix obtained as a result of the sample preparation steps was. Conclusion: As a result, a fast, inexpensive LC-MS/MS method has been developed for 3-Phenylpyruvic acid.
Açıklama
Anahtar Kelimeler
3-phenylpyruvic acid, trans cinnamic acid, LC-MS/MS, phenylketonuria, liquid-liquid extraction.
Kaynak
Ege Tıp Dergisi
WoS Q Değeri
Scopus Q Değeri
Cilt
63
Sayı
1
