Unexpected retention behavior of baicalin: Hydrophilic interaction like properties of a reversed-phase column

Balázs Magda, Zoltán Márta, T. Imre, Bernadett Kalapos-Kovács, I. Klebovich, J. Fekete, P. Szabó

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The original aim of this study was to develop a method for the determination of baicalin from membrane vesicles. The unconventional chromatographic separation ("inverse gradient elution" on a reversed phase column) was due to a lucky chance, which is detailed and discussed in this study. The validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is proved to be sensitive, rapid and selective. Chromatographic separation was performed on a Zorbax SB-C8 column (250. mm. ×. 4.6. mm, i.d.; 5. μm) with 0.1% formic acid in water and methanol by linear gradient elution. Quantification of baicalin was determined by multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The calibration curve was linear (r= 0.9987) over the concentration range from 1 to 1000. nM. The coefficient of variation and relative error of baicalin for intra- and inter-assay at three quality control (QC) levels were 2.0-10.2% and -6.1 to 6.7%, respectively. The lower limit of quantification (LLOQ) for baicalin was 1. nM (0.446. ng/ml), without preconcentration of the sample. This method was subsequently applied to vesicular transport assays of baicalin in membrane vesicles successfully. The developed method can open up new area of research in the chromatographic separation of flavonoids and their glucuronides.

Original languageEnglish
Pages (from-to)119-125
Number of pages7
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume111
DOIs
Publication statusPublished - Jul 1 2015

Fingerprint

Hydrophobic and Hydrophilic Interactions
Assays
formic acid
Membranes
Electrospray ionization
Glucuronides
Liquid chromatography
Tandem Mass Spectrometry
Flavonoids
Liquid Chromatography
Quality Control
Calibration
Quality control
Mass spectrometry
Methanol
baicalin
Water
Monitoring
Research

Keywords

  • Baicalin
  • Hydrophilic interaction chromatography
  • LC-MS/MS
  • Reversed-phase chromatography
  • Vesicular transport assay

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmaceutical Science
  • Spectroscopy
  • Clinical Biochemistry

Cite this

Unexpected retention behavior of baicalin : Hydrophilic interaction like properties of a reversed-phase column. / Magda, Balázs; Márta, Zoltán; Imre, T.; Kalapos-Kovács, Bernadett; Klebovich, I.; Fekete, J.; Szabó, P.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 111, 01.07.2015, p. 119-125.

Research output: Contribution to journalArticle

@article{3b13915f17274efb9d43c7d76849deb4,
title = "Unexpected retention behavior of baicalin: Hydrophilic interaction like properties of a reversed-phase column",
abstract = "The original aim of this study was to develop a method for the determination of baicalin from membrane vesicles. The unconventional chromatographic separation ({"}inverse gradient elution{"} on a reversed phase column) was due to a lucky chance, which is detailed and discussed in this study. The validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is proved to be sensitive, rapid and selective. Chromatographic separation was performed on a Zorbax SB-C8 column (250. mm. ×. 4.6. mm, i.d.; 5. μm) with 0.1{\%} formic acid in water and methanol by linear gradient elution. Quantification of baicalin was determined by multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The calibration curve was linear (r= 0.9987) over the concentration range from 1 to 1000. nM. The coefficient of variation and relative error of baicalin for intra- and inter-assay at three quality control (QC) levels were 2.0-10.2{\%} and -6.1 to 6.7{\%}, respectively. The lower limit of quantification (LLOQ) for baicalin was 1. nM (0.446. ng/ml), without preconcentration of the sample. This method was subsequently applied to vesicular transport assays of baicalin in membrane vesicles successfully. The developed method can open up new area of research in the chromatographic separation of flavonoids and their glucuronides.",
keywords = "Baicalin, Hydrophilic interaction chromatography, LC-MS/MS, Reversed-phase chromatography, Vesicular transport assay",
author = "Bal{\'a}zs Magda and Zolt{\'a}n M{\'a}rta and T. Imre and Bernadett Kalapos-Kov{\'a}cs and I. Klebovich and J. Fekete and P. Szab{\'o}",
year = "2015",
month = "7",
day = "1",
doi = "10.1016/j.jpba.2015.03.037",
language = "English",
volume = "111",
pages = "119--125",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
issn = "0731-7085",
publisher = "Elsevier",

}

TY - JOUR

T1 - Unexpected retention behavior of baicalin

T2 - Hydrophilic interaction like properties of a reversed-phase column

AU - Magda, Balázs

AU - Márta, Zoltán

AU - Imre, T.

AU - Kalapos-Kovács, Bernadett

AU - Klebovich, I.

AU - Fekete, J.

AU - Szabó, P.

PY - 2015/7/1

Y1 - 2015/7/1

N2 - The original aim of this study was to develop a method for the determination of baicalin from membrane vesicles. The unconventional chromatographic separation ("inverse gradient elution" on a reversed phase column) was due to a lucky chance, which is detailed and discussed in this study. The validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is proved to be sensitive, rapid and selective. Chromatographic separation was performed on a Zorbax SB-C8 column (250. mm. ×. 4.6. mm, i.d.; 5. μm) with 0.1% formic acid in water and methanol by linear gradient elution. Quantification of baicalin was determined by multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The calibration curve was linear (r= 0.9987) over the concentration range from 1 to 1000. nM. The coefficient of variation and relative error of baicalin for intra- and inter-assay at three quality control (QC) levels were 2.0-10.2% and -6.1 to 6.7%, respectively. The lower limit of quantification (LLOQ) for baicalin was 1. nM (0.446. ng/ml), without preconcentration of the sample. This method was subsequently applied to vesicular transport assays of baicalin in membrane vesicles successfully. The developed method can open up new area of research in the chromatographic separation of flavonoids and their glucuronides.

AB - The original aim of this study was to develop a method for the determination of baicalin from membrane vesicles. The unconventional chromatographic separation ("inverse gradient elution" on a reversed phase column) was due to a lucky chance, which is detailed and discussed in this study. The validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is proved to be sensitive, rapid and selective. Chromatographic separation was performed on a Zorbax SB-C8 column (250. mm. ×. 4.6. mm, i.d.; 5. μm) with 0.1% formic acid in water and methanol by linear gradient elution. Quantification of baicalin was determined by multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The calibration curve was linear (r= 0.9987) over the concentration range from 1 to 1000. nM. The coefficient of variation and relative error of baicalin for intra- and inter-assay at three quality control (QC) levels were 2.0-10.2% and -6.1 to 6.7%, respectively. The lower limit of quantification (LLOQ) for baicalin was 1. nM (0.446. ng/ml), without preconcentration of the sample. This method was subsequently applied to vesicular transport assays of baicalin in membrane vesicles successfully. The developed method can open up new area of research in the chromatographic separation of flavonoids and their glucuronides.

KW - Baicalin

KW - Hydrophilic interaction chromatography

KW - LC-MS/MS

KW - Reversed-phase chromatography

KW - Vesicular transport assay

UR - http://www.scopus.com/inward/record.url?scp=84928535185&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84928535185&partnerID=8YFLogxK

U2 - 10.1016/j.jpba.2015.03.037

DO - 10.1016/j.jpba.2015.03.037

M3 - Article

C2 - 25880242

AN - SCOPUS:84928535185

VL - 111

SP - 119

EP - 125

JO - Journal of Pharmaceutical and Biomedical Analysis

JF - Journal of Pharmaceutical and Biomedical Analysis

SN - 0731-7085

ER -