여름정기학술대회
2022여름초록
발표자 및 발표 내용
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| Brief Oral Presentation 발표신청 |
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주저자
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공동저자
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접수자
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The
endogenous amino acids (AAs) level can reflect biological metabolism and
function including diseases and damage. The liver plays a key role in
determining drugs toxicity due to its role in body metabolism, and the AAs
level in liver is a candidate biomarker for monitoring of drug-induced liver
injury (DILI). In this study, isotope dilution liquid chromatography-mass
spectrometry (ID-LC-MS) based methods for seventeen AAs (proline, valine, leucine,
phenylalanine, isoleucine, tyrosine, histidine, methionine, aspartic acid, threonine,
arginine, tryptophan, glutamic acid, glutamine, alanine, glycine, and serine) were
described for liver organoids with DILI. Two methods with isotopes as internal
standards were; 1) ID-LC-MS for underivatized AAs via deproteinization with 15
v/v% 5-sulfosalicylic acid, and 2) ID-LC-MS for derivatized AAs via
deproteinization with 75 v/v% acetonitrile and derivatization by AccQ-Tag™ kit.
The measurement procedures were optimized and verified by using available
certified reference materials with claimed certified values for AAs in plasma. All target AA residues were clearly separated within 10 min
in both LC-MS conditions. The measurement procedures were applied to liver
organoids. The liver organoids were treated with drugs (Troglitazone, Ketoconazole,
Diclofenac, and Benzbromarone) for DILI, and the AA contents in both cell
lysate and media were quantified and normalized by protein concentration. Particularly,
the levels of glutamine, glutamic acid, aspartic acid, and tryptophan were
significantly different from DILI groups comparing with control group. The AA
contents in the cell media also showed different pattern between DILI and control
groups. These results indicate that AAs can be effective biomarkers for
drug-induced hepatotoxicity. Moreover, the optimized LC-MS
methods will be suitable for the accurate and efficient quantification method
for AA profiling in various samples.
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