Identification of a major metabolite of danshensu in rat urine and simultaneous determination of danshensu and its metabolite in plasma: application to a pharmacokinetic study in rats

Abstract

Danshensu, as the effective component of Salvia miltiorrhiza (Danshen), has been widely used in clinical studies for treatment of cardiovascular diseases in China. A new metabolite, 4-hydroxy-3-methoxyphenyllactic acid was isolated from the urine of rats, and its chemical structure was identified by ultraviolet (UV), Infrared Spectroscopy (IR), mass spectrometry (MS) and nuclear magnetic resonance (NMR). Furthermore, a selective and sensitive high performance liquid chromatography-tandem mass spectrometric (HPLC-MS/MS) method was developed for the simultaneous quantification of danshensu and its major metabolite, 4-hydroxy-3-methoxyphenyllactic acid, in rat plasma after oral and intravenous administration of danshensu. The separation was performed on a Hypersil Gold C18 column (150 × 2.1 mm i.d., 3.0 µm, Thermo, San jose CA, USA) with gradient elution using a mobile phase composed of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. Linear detection responses were obtained for danshensu and 4-hydroxy-3-methoxyphenyllactic acid ranging from 5 to 10000 ng/mL and 5 to 4000 ng/mL, respectively. The lower limits of quantification (LLOQs) for the two compounds were both 5 ng/mL. The intra-and inter-day precision (R.S.D %) were within 5.61% for the two analytes. The average recoveries of the analytes were greater than 72.43%. The method was proved to be stable during all sample storage, preparation and analytic procedures. This method was successfully applied to the pharmacokinetic studies of danshensu and 4-hydroxy-3-methoxyphenyllactic acid after oral and intravenous administration of danshensu in rats.

Metabolite identification of tanshinol borneol ester in rats by high‐performance liquid chromatography combined with electrospray ionization quadrupole time‐of‐flight mass spectrometry

“…These m / z values were 14 mass units higher than those of the parent drug M0, indicating that they were the O‐ methylated DBZ. In addition, the product ion at m / z 211.0603 was consistent with the quasi‐molecular ion of O‐ methylated DSS, according to a previous report (Wang et al, ). The ion at m / z 165.0578 was generated via the loss of HCOOH from the ion at m / z 211.0603 (Yang et al, ), while the fragment ion at m / z 153.1277 was tentatively supposed to be 1 mole of borneol.…”

“…DSS is a predominant bioactive component in aqueous extract of Salvia miltiorrhiza , which is the Jun (emperor) medicinal of FFDS (Zhao et al, ). It can delay the senescence of rat aortic endothelial cells (Wang et al, ), inhibit platelet aggregation and thrombus formation (Yu et al, ), and protect against cardio‐cerebral ischaemia/reperfusion injury (Fan et al, ; Guo et al, ; Meng et al, ; Yin et al, ). In addition to the above‐mentioned effects, DSS has also been reported to inhibit β ‐adrenergic receptor‐mediated cardiac fibrosis and carbon tetrachloride‐induced hepatic fibrosis (Lu et al, ; Qu, Huang, Li, & Qin, ).…”

Identification of metabolites of isopropyl 3‐(3,4‐dihydroxyphenyl)‐2‐hydroxypropanoate in rats by high‐performance liquid chromatography combined with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry

“…Among these methods, the application of HPLC‐UV (Vella et al ., ), HPLC‐ECD (Jia et al ., ) and HPLC‐FLD (Zhao et al ., ) is limited by the use of reference standards and the lack of high‐throughput qualitation of unknown compounds. In contrast, high‐performance liquid chromatography combined with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry (HPLC‐ESI‐Q‐TOF‐MS; Wang et al ., ) has become increasingly attractive for rapid screening and characterization of known or unknown drug metabolites in biological matrix, owing to its fast online separation, excellent detection sensitivity and great accuracy (Liu et al, ). It can provide accurate mass measurement and elemental composition assignment which are essential for the characterization of nontarget compounds (Gu et al ., ).…”

Recommendations on the Clinical Use of Compound Danshen Dripping Pills

“…[23456] Following the administration of clinical and nonclinical studies of CDDPs to either rats or healthy volunteers, Tanshinol and its main metabolite 4-hydroxy-3-methoxylphenyllactic acid,[3] protocatechualdehyde and its main metabolite protocatechuic acid,[4] notoginsenoside R 1 , and ginsenosides Rg 1 and Rb 1 [5] were detectable in the blood samples. Among those, T max for tanshinol, metabolite 4-hydroxy-3-methoxylphenyllactic acid, protocatechuic acid, notoginsenoside R 1 , and ginsenoside Rg 1 were all <1 h, indicating that CDDP was absorbed very rapidly after its oral administration, while a moderate absorption rate for ginsenoside Rg 1 was observed with a relatively longer half-life, maintaining a sustainable clinical effectiveness.…”