|
Type |
Oral Presentation |
Area |
Oral Presentation of Young Analytical Chemists Ⅱ |
Room No. |
303호 |
Time |
FRI 09:46-: |
Code |
ANAL2.O-20 |
Subject |
iCCM-based isotope dilution mass spectrometry for absolute quantification of human growth hormone and brain natriuretic peptide-32 in human plasma |
Authors |
이선영, 홍종기, 강덕진1,* 경희대학교 약학과, Korea 1한국표준과학연구원(KRISS) 삶의질측정표준본부, Korea |
Abstract |
Isotope dilution mass spectrometry (ID-MS) has been used as a primary method for the absolute quantitative determination. Although the conventionally absolute quantification (e.g., synthetic proteins or peptides) or stable isotope labeling methods (e.g., ICAT, mTRAQ) in quantitative proteomics have been considered as a reliable approach in shotgun proteomics, these methods have unfortunately several bottlenecks such as imperfect labeling, and time consumption of labeling for proteome samples. To improve above-mentioned problems, we applied the carbamidomethylation-based isotope labeling (iCCM) for absolute quantification of hGH and BNP-32 in human plasma in this study. To evaluate the iCCM labeling for absolute quantification, plasma sample was first labeled with iodoacetamide (IAA) as carbamidomethylation (CM), while both human growth hormone (hGH) and brain natriuretic peptide-32 (BNP-32) standards were isotopically alkylated with IAA isotope (IAA-13C2D2) as iCCM, followed by spiking those iCCM-labeled standards to plasma sample labeled with CM. The resulting plasma sample was directly performed tryptic digestion and subsequently applied to nLC-ESI-MS/MS analysis so as to obtain the absolute amounts of both hGH and BNP-32. In conclusion, iCCM-based ID-MS provides the simple, rapid, and high reproductive method for the assessment of the ratio of CM/iCCM, resulting in absolute quantification of hGH and BNP-32. |
E-mail |
sylee0109@kriss.re.kr |
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