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A strategy for integrated pharmacokinetic study of cardiovascular herbal medicines based on chemiluminescence and HPLC-MS/MS assays: a case using Danshen injection

Yao, Hong, Shi, Peiying, Huang, Xiaomei, Shen, Zhenhuang, Su, Ya, Yang, Chunlei, Zhu, Meilan, Huang, Liying, Liu, Ailin, Lin, Xinhua
RSC advances 2017 v.7 no.22 pp. 13570-13583
Oriental traditional medicine, aldehydes, antioxidants, chemiluminescence, danshensu, herbal medicines, high performance liquid chromatography, ingredients, mice, monitoring, pharmacokinetics, protective effect, rats, rosmarinic acid, salvianolic acid, sodium, tandem mass spectrometry, therapeutics
Currently, it is important to explore any feasible tool to represent the integrated in vivo process of a traditional Chinese medicine (TCM) to understand its clinical dosage regimen from ancient empirical therapy. Herein, we put forward a strategy to characterize the integrated pharmacokinetic properties of multiple-component cardiovascular herbal medicines with a flow injection chemiluminescence (FI-CL) technique and an HPLC-MS/MS assay. Taking Danshen injection as a carrier, the main ingredients of this preparation were determined by HPLC as danshensu sodium (DSSN, 554.13 mg g⁻¹), protocatechuic aldehyde (PCAL, 40.55 mg g⁻¹), rosmarinic acid (RA, 21.66 mg g⁻¹), salvianolic acid B (SAB, 1.67 mg g⁻¹) and salvianolic acid A (SAA, 68.60 mg g⁻¹). This was followed by confirmation that these phenolics could inhibit the CL signal of the KIO₄–luminol system and that the inhibition was closely correlative with both their concentrations and antioxidative capability. Further, the protective effects of the main components (40 mg kg⁻¹ per day, i.v. for 7 days) on isoprenaline-induced myocardial injury in mice were evaluated, confirming that these phenolics were the main effective substances and that antioxidation was the most important of their therapeutic effects. Subsequently, the PK profiles of the main phenolic acids in rat plasma (21 mg kg⁻¹, single i.v.) were studied by HPLC-MS/MS with a multiple reaction monitoring mode. The phenolic acid mixture solutions could be prepared in distilled water according to their plasma drug concentrations to simulate the plasma sample solutions at each studied PK time point. All these provided the basis to evaluate their integrated PK properties through determining the total phenolic acid content in the simulation solutions by the inhibited CL signal with salvianolic acid A (SAA), the most powerful ingredient in Danshen injection, as a reference. Finally, the integrated PK properties can be characterized into the SAA “equivalent” parameters, including t₁/₂ᵦ, k, and AUC₍₀–∞₎, by the inhibited CL signal. Also, the results were compared with those from the “plasma drug concentration sum method” and the “AUC weighting integrated method” from previous reports. In summary, the present study provides a feasible strategy for PK analysis of cardiovascular herbal medicines with antioxidants as the main effective substances.