A Fast & Reliable Liquid Chromatography Mass Spectrometry Method for Determination of Amino acids & Acylcarnitines including Succinyl acetone in dried blood spots for newborn screening.
|International Journal of Medical Science|
|© 2019 by SSRG - IJMS Journal|
|Volume 6 Issue 7|
|Year of Publication : 2019|
|Authors : Shrimant N Panaskar, Mahesh H Hampe, Mr. Susheel Singh, Akshata Shivalkar, Pallavi Kadam|
How to Cite?
Shrimant N Panaskar, Mahesh H Hampe, Mr. Susheel Singh, Akshata Shivalkar, Pallavi Kadam, "A Fast & Reliable Liquid Chromatography Mass Spectrometry Method for Determination of Amino acids & Acylcarnitines including Succinyl acetone in dried blood spots for newborn screening.," SSRG International Journal of Medical Science, vol. 6, no. 7, pp. 1-11, 2019. Crossref, https://doi.org/10.14445/23939117/IJMS-V6I7P101
Introduction:We developed & validate first time TMS based non derivatized 1.5 min protocol to quantify 25 amino acids & 30 acylcarnitines along with Succinyl acetone.
Method: 3.2mm DBS disk was extracted with a mixture of methanol: Hydrate hydrazine: oxalic acid: Acetonitrile: Water. Analysis was performed on LCMSMS with direct injection method. Water: formic acid & Methanol was used in gradient elution. Optimized multiple reaction monitoring (MRM) was used for detection of amino acids and acylcarnitines on a Shimatzu mass spectrometer. The method was validated with respect to linearity, precision, recovery, accuracy.
Result: Precision, accuracy and analytical measurement range considering overall precision within 10% and accuracy within 15% of for all amino acids for Acylcarntine at one fortified level, which quantitated 21.8% lower than the expected value. Inter method comparison shows only 8.5% & 10.5 % error bias for AA & AC respectively. Analytes were stable for 1 month at room temperature & 6 months at -200C storage condition.
Conclusion: The present design protocol is sensitive with significant LOD and cost effective which can be effectively applied as fast primary & second-tier test for newborn screening of metabolic disorders associated with abnormal levels of acylcarnitines and amino acids, potentially reducing false positive cases and shortening the time to diagnosis.
Amino acids & Acylcarnitines, Dried blood spot, ESI-MS, newborn screening
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