Limits...
A MALDI-MS-based quantitative analytical method for endogenous estrone in human breast cancer cells.

Kim KJ, Kim HJ, Park HG, Hwang CH, Sung C, Jang KS, Park SH, Kim BG, Lee YK, Yang YH, Jeong JH, Kim YG - Sci Rep (2016)

Bottom Line: The one-step modification of endogenous estrone provided good linearity (R(2) > 0.99) and significantly increased the sensitivity of the platform (limit of quantitation: 11 fmol).Finally, by applying the MALDI-MS-based quantitative method to endogenous estrones, we successfully monitored changes in the metabolic expression level of estrones (17.7 fmol/10(6) letrozole-treated cells) in MCF-7 cells resulting from treatment with an aromatase inhibitor.Taken together, these results suggest that this MALDI-MS-based quantitative approach may be a general method for the targeted metabolomics of ketone-containing metabolites, which can reflect clinical conditions and pathogenic mechanisms.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Engineering, Soongsil University, Seoul 156-743, Korea.

ABSTRACT
The level of endogenous estrone, one of the three major naturally occurring estrogens, has a significant correlation with the incidence of post-menopausal breast cancer. However, it is challenging to quantitatively monitor it owing to its low abundance. Here, we develop a robust and highly sensitive mass-assisted laser desorption/ionization mass spectrometry (MALDI-MS)-based quantitative platform to identify the absolute quantities of endogenous estrones in a variety of clinical specimens. The one-step modification of endogenous estrone provided good linearity (R(2) > 0.99) and significantly increased the sensitivity of the platform (limit of quantitation: 11 fmol). In addition, we could identify the absolute amount of endogenous estrones in cells of the breast cancer cell line MCF-7 (34 fmol/10(6) cells) by using a deuterated estrone as an internal standard. Finally, by applying the MALDI-MS-based quantitative method to endogenous estrones, we successfully monitored changes in the metabolic expression level of estrones (17.7 fmol/10(6) letrozole-treated cells) in MCF-7 cells resulting from treatment with an aromatase inhibitor. Taken together, these results suggest that this MALDI-MS-based quantitative approach may be a general method for the targeted metabolomics of ketone-containing metabolites, which can reflect clinical conditions and pathogenic mechanisms.

No MeSH data available.


Related in: MedlinePlus

Schematic strategy of the MALDI-MS-based quantitative analysis of ketone-containing endogenous estrones.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4836303&req=5

f1: Schematic strategy of the MALDI-MS-based quantitative analysis of ketone-containing endogenous estrones.

Mentions: To address these problems, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), which is an extremely sensitive and robust method that is suitable for the analysis of complex mixtures, could be a great alternative tool for the analysis of endogenous estrones3435. Here, we demonstrate a MALDI-based quantitative platform for the quantitation of endogenous estrogenic hormones in human body fluid and cells (Fig. 1). First, we introduced a permanent cationic charge onto the ketone-containing E1 to simplify the MALDI spectra due to alkali metal adducts (e.g., Li+, Na+, or K+). The reliability of the quantitative method was validated using synthetic ketone-containing steroid hormones (i.e., estrone and testosterone). After the chemical derivatization, there are significant improvements in the signal reproducibility and linearity (R2 > 0.99) compared to the underivatized counterparts. We used deuterated estrone (d4-E1) as an internal standard to monitor the change in the absolute amount of endogenous estrones in MCF-7 cells resulting from treatment with an aromatase inhibitor.


A MALDI-MS-based quantitative analytical method for endogenous estrone in human breast cancer cells.

Kim KJ, Kim HJ, Park HG, Hwang CH, Sung C, Jang KS, Park SH, Kim BG, Lee YK, Yang YH, Jeong JH, Kim YG - Sci Rep (2016)

Schematic strategy of the MALDI-MS-based quantitative analysis of ketone-containing endogenous estrones.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836303&req=5

f1: Schematic strategy of the MALDI-MS-based quantitative analysis of ketone-containing endogenous estrones.
Mentions: To address these problems, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), which is an extremely sensitive and robust method that is suitable for the analysis of complex mixtures, could be a great alternative tool for the analysis of endogenous estrones3435. Here, we demonstrate a MALDI-based quantitative platform for the quantitation of endogenous estrogenic hormones in human body fluid and cells (Fig. 1). First, we introduced a permanent cationic charge onto the ketone-containing E1 to simplify the MALDI spectra due to alkali metal adducts (e.g., Li+, Na+, or K+). The reliability of the quantitative method was validated using synthetic ketone-containing steroid hormones (i.e., estrone and testosterone). After the chemical derivatization, there are significant improvements in the signal reproducibility and linearity (R2 > 0.99) compared to the underivatized counterparts. We used deuterated estrone (d4-E1) as an internal standard to monitor the change in the absolute amount of endogenous estrones in MCF-7 cells resulting from treatment with an aromatase inhibitor.

Bottom Line: The one-step modification of endogenous estrone provided good linearity (R(2) > 0.99) and significantly increased the sensitivity of the platform (limit of quantitation: 11 fmol).Finally, by applying the MALDI-MS-based quantitative method to endogenous estrones, we successfully monitored changes in the metabolic expression level of estrones (17.7 fmol/10(6) letrozole-treated cells) in MCF-7 cells resulting from treatment with an aromatase inhibitor.Taken together, these results suggest that this MALDI-MS-based quantitative approach may be a general method for the targeted metabolomics of ketone-containing metabolites, which can reflect clinical conditions and pathogenic mechanisms.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Engineering, Soongsil University, Seoul 156-743, Korea.

ABSTRACT
The level of endogenous estrone, one of the three major naturally occurring estrogens, has a significant correlation with the incidence of post-menopausal breast cancer. However, it is challenging to quantitatively monitor it owing to its low abundance. Here, we develop a robust and highly sensitive mass-assisted laser desorption/ionization mass spectrometry (MALDI-MS)-based quantitative platform to identify the absolute quantities of endogenous estrones in a variety of clinical specimens. The one-step modification of endogenous estrone provided good linearity (R(2) > 0.99) and significantly increased the sensitivity of the platform (limit of quantitation: 11 fmol). In addition, we could identify the absolute amount of endogenous estrones in cells of the breast cancer cell line MCF-7 (34 fmol/10(6) cells) by using a deuterated estrone as an internal standard. Finally, by applying the MALDI-MS-based quantitative method to endogenous estrones, we successfully monitored changes in the metabolic expression level of estrones (17.7 fmol/10(6) letrozole-treated cells) in MCF-7 cells resulting from treatment with an aromatase inhibitor. Taken together, these results suggest that this MALDI-MS-based quantitative approach may be a general method for the targeted metabolomics of ketone-containing metabolites, which can reflect clinical conditions and pathogenic mechanisms.

No MeSH data available.


Related in: MedlinePlus