Determination of Cryptotanshinone, a Major Active Component in Salvia miltiorrhiza, in Caco-2 Monolayers by HPLC with UV Detection

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.