Determination of cryptotanshinone, a major active component in Salvia miltiorrhiza, in Caco-2 monolayers by HPLC with UV detection

Su Guan, Hui Chang Bi, Ying Pan, Xiao Chen, Zhi Wei Zhou, Charlie Changli Xue, Chun Guang Li, Min Huang, Shu Feng Zhou

Research output: Contribution to journalArticlepeer-review

Abstract

Cryptotanshinone is the major active component from the root of Salvia miltiorrhiza which has been widely used for the management of coronary heart disease. The aims of this study were to develop and validate an HPLC method for the determination of cryptotanshinone in the human intestinal cell line Caco-2 monolayers, and to investigate the transport kinetics of cryptotanshinone. The developed HPLC method was sensitive and reliable, with acceptable accuracy (90-110% of true values) and precision (intra- and inter-assay CV < 10%). The total running time was within 10 min, with acceptable separation of the compounds of interest. The limit of quantitation (LOQ) for cryptotanshinone was 10 ng mL-1. A simple liquid-liquid extraction procedures resulted in an extraction efficiency of 90.8 ± 8.9 and 93.5 ± 6.2% for cryptotanshinone at 0.1 and 3 μg mL-1. The calibration curve was linear over the concentration range of 0.05-3.0 μg mL-1 with the mean correlation coefficients >0.999. The validated HPLC method was applied to examine the epithelial transport of cryptotanshinone by Caco-2 monolayers. The transport across the monolayers from the apical (B) to basolateral (A) side was significantly higher than that from A to B side. The structural analog of cryptotanshinone and a known substrate of P-glycoprotein, tanshinone II A, dramatically inhibited the B to A transport of cryptotanshinone in the monolayers. These results indicate that the developed HPLC method was suitable for the study of transport of cryptotanshinone by Caco-2 monolayers and cryptotanshinone is a substrate of P-glycoprotein.

Original languageEnglish (US)
Pages (from-to)577-582
Number of pages6
JournalChromatographia
Volume66
Issue number7-8
DOIs
StatePublished - Oct 2007
Externally publishedYes

Keywords

  • Coca-2, transport
  • Column liquid chromatography
  • Cryptotanshinone
  • Validation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Organic Chemistry

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