Comparison of partial filling MEKC analyses of steroids with use of ESI‐MS and UV spectrophotometry

Lotta Amundsen, Juha T. Kokkonen, Heli Sirén (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

18 Citations (Scopus)

Abstract

Until now, partial filling micellar EKC–ESI‐MS (PF‐MEKC–ESI‐MS) has seldom been applied for human biomolecules. In this study, determination of three electrically neutral endogenous steroid hormones, namely androstenedione, testosterone, and epitestosterone, by PF‐MEKC–ESI‐MS was investigated. Since ESI‐MS and ESI‐MS/MS behaviors of testosterone and epitestosterone proved to be nearly identical, efficient separation of the two compounds was required to obtain reliable identification. The chemical conditions as well as the instrumental parameters affecting the PF‐MEKC–ESI‐MS analysis were researched. In optimal conditions, ESI‐MS showed excellent selectivity, and all the steroids could be identified using SIM. LODs were 0.75–5 μg/mL. The results obtained by PF‐MEKC–ESI‐MS were compared with those obtained by corresponding PF‐MEKC–UV. PF‐MEKC–UV provided better resolution, repeatability, and more than ten‐fold higher sensitivity, in terms of LODs, than PF‐MEKC–ESI‐MS. The reasons for this were carefully examined. In comparison with PF‐MEKC–UV, PF‐MEKC–ESI‐MS suffered from greater band broadening owing to the sheath–liquid interface. Resolution was also decreased owing to the elevated capillary temperature. Finally, we discovered that in the analysis of electrically neutral compounds, in‐capillary sample concentration by micellar sweeping could be more efficiently utilized in PF‐MEKC–UV than in PF‐MEKC–ESI‐MS.
Original languageEnglish
Pages (from-to)803-813
JournalJournal of Separation Science
Volume31
Issue number5
DOIs
Publication statusPublished - 2008
MoE publication typeA1 Journal article-refereed

Fingerprint

Spectrophotometry
Steroids
Epitestosterone
Testosterone
Steroid hormones
Androstenedione
Biomolecules

Keywords

  • testosterone
  • epitestosterone
  • steroid
  • partial filling micellar electrokinetic chromatography
  • electrospray ionization mass spectrometry

Cite this

Amundsen, Lotta ; Kokkonen, Juha T. ; Sirén, Heli. / Comparison of partial filling MEKC analyses of steroids with use of ESI‐MS and UV spectrophotometry. In: Journal of Separation Science. 2008 ; Vol. 31, No. 5. pp. 803-813.
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abstract = "Until now, partial filling micellar EKC–ESI‐MS (PF‐MEKC–ESI‐MS) has seldom been applied for human biomolecules. In this study, determination of three electrically neutral endogenous steroid hormones, namely androstenedione, testosterone, and epitestosterone, by PF‐MEKC–ESI‐MS was investigated. Since ESI‐MS and ESI‐MS/MS behaviors of testosterone and epitestosterone proved to be nearly identical, efficient separation of the two compounds was required to obtain reliable identification. The chemical conditions as well as the instrumental parameters affecting the PF‐MEKC–ESI‐MS analysis were researched. In optimal conditions, ESI‐MS showed excellent selectivity, and all the steroids could be identified using SIM. LODs were 0.75–5 μg/mL. The results obtained by PF‐MEKC–ESI‐MS were compared with those obtained by corresponding PF‐MEKC–UV. PF‐MEKC–UV provided better resolution, repeatability, and more than ten‐fold higher sensitivity, in terms of LODs, than PF‐MEKC–ESI‐MS. The reasons for this were carefully examined. In comparison with PF‐MEKC–UV, PF‐MEKC–ESI‐MS suffered from greater band broadening owing to the sheath–liquid interface. Resolution was also decreased owing to the elevated capillary temperature. Finally, we discovered that in the analysis of electrically neutral compounds, in‐capillary sample concentration by micellar sweeping could be more efficiently utilized in PF‐MEKC–UV than in PF‐MEKC–ESI‐MS.",
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Comparison of partial filling MEKC analyses of steroids with use of ESI‐MS and UV spectrophotometry. / Amundsen, Lotta; Kokkonen, Juha T.; Sirén, Heli (Corresponding Author).

In: Journal of Separation Science, Vol. 31, No. 5, 2008, p. 803-813.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Comparison of partial filling MEKC analyses of steroids with use of ESI‐MS and UV spectrophotometry

AU - Amundsen, Lotta

AU - Kokkonen, Juha T.

AU - Sirén, Heli

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N2 - Until now, partial filling micellar EKC–ESI‐MS (PF‐MEKC–ESI‐MS) has seldom been applied for human biomolecules. In this study, determination of three electrically neutral endogenous steroid hormones, namely androstenedione, testosterone, and epitestosterone, by PF‐MEKC–ESI‐MS was investigated. Since ESI‐MS and ESI‐MS/MS behaviors of testosterone and epitestosterone proved to be nearly identical, efficient separation of the two compounds was required to obtain reliable identification. The chemical conditions as well as the instrumental parameters affecting the PF‐MEKC–ESI‐MS analysis were researched. In optimal conditions, ESI‐MS showed excellent selectivity, and all the steroids could be identified using SIM. LODs were 0.75–5 μg/mL. The results obtained by PF‐MEKC–ESI‐MS were compared with those obtained by corresponding PF‐MEKC–UV. PF‐MEKC–UV provided better resolution, repeatability, and more than ten‐fold higher sensitivity, in terms of LODs, than PF‐MEKC–ESI‐MS. The reasons for this were carefully examined. In comparison with PF‐MEKC–UV, PF‐MEKC–ESI‐MS suffered from greater band broadening owing to the sheath–liquid interface. Resolution was also decreased owing to the elevated capillary temperature. Finally, we discovered that in the analysis of electrically neutral compounds, in‐capillary sample concentration by micellar sweeping could be more efficiently utilized in PF‐MEKC–UV than in PF‐MEKC–ESI‐MS.

AB - Until now, partial filling micellar EKC–ESI‐MS (PF‐MEKC–ESI‐MS) has seldom been applied for human biomolecules. In this study, determination of three electrically neutral endogenous steroid hormones, namely androstenedione, testosterone, and epitestosterone, by PF‐MEKC–ESI‐MS was investigated. Since ESI‐MS and ESI‐MS/MS behaviors of testosterone and epitestosterone proved to be nearly identical, efficient separation of the two compounds was required to obtain reliable identification. The chemical conditions as well as the instrumental parameters affecting the PF‐MEKC–ESI‐MS analysis were researched. In optimal conditions, ESI‐MS showed excellent selectivity, and all the steroids could be identified using SIM. LODs were 0.75–5 μg/mL. The results obtained by PF‐MEKC–ESI‐MS were compared with those obtained by corresponding PF‐MEKC–UV. PF‐MEKC–UV provided better resolution, repeatability, and more than ten‐fold higher sensitivity, in terms of LODs, than PF‐MEKC–ESI‐MS. The reasons for this were carefully examined. In comparison with PF‐MEKC–UV, PF‐MEKC–ESI‐MS suffered from greater band broadening owing to the sheath–liquid interface. Resolution was also decreased owing to the elevated capillary temperature. Finally, we discovered that in the analysis of electrically neutral compounds, in‐capillary sample concentration by micellar sweeping could be more efficiently utilized in PF‐MEKC–UV than in PF‐MEKC–ESI‐MS.

KW - testosterone

KW - epitestosterone

KW - steroid

KW - partial filling micellar electrokinetic chromatography

KW - electrospray ionization mass spectrometry

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