The Pharmacokinetic Studies Of Volatile Compounds Of Shexiang Baoxin Pill

Shexiang Baoxin Pill (SBP) is one of the classic formula which has been commonly used in clinical practice to treat cardiovascular disease, such as coronary heart diseases, and angina pectoris. SBP composed of seven herbs including Borneolum Syntheticum, Moschus, Venenum Bufonis, Radix Ginseng, Calculus Bovis, Cortex Cinnamomi, and Styrax. In our previous study, the serum pharmaceutical chemistry shown that SBP contain30compounds (22prototypes and8metabolites) in plasma, such as muscone, cinnamaldehyde, cinnamic acid, ginsenoside, cardiotonic steroids, cholic acid, and so on. In SBP, four of its seven medicinal materials (Borneolum Syntheticum, Moschus, Styrax and Cortex Cinnamomi), contain a deal of volatile compounds such as isoborneol, borneol, muscone, cinnamaldehyde, cinnamic acid and benzyl benzoate. Borneolum Syntheticum and Moschus has shown some pharmacological properties including cardiovascular, vasodilative, analgesic and anti-inflammatory. Styrax has been considered to show a therapeutic effect on ischemia-reperfusion injuryin cardiac myocytes. Cortex Cinnamomi displays vasodilative and angiogenic effects. The above studies suggested that the volatile components, in the same way, are the major bioactive ingredients of SBP, which may be responsible for its therapeutic efficacy. However, the pharmacokinetic properties of volatile compounds of SBP remain challenging due to the low concentration in plasma and large losses during the sample preparation.In our former study, the serum pharmaceutical chemistry and assay of six volatile compounds of SBP have been accomplished by GC-MS. The results shown that isoborneol, borneol, muscone, cinnamaldehyde, cinnamic acid and benzyl benzoate were the main compounds in plasma. Based on the result of serum pharmaceutical chemistry, the headspace solid-phase dynamic extraction method coupled to gas chromatography-tandem mass spectrometry (HS-SPDE-GC-MS/MS) of simultaneous determination of isoborneol, borneol, muscone and cinnamaldehyde, and its application to pharmacokinetic studies after oral administration of SBP in rats were studied in this paper.The target compounds were extracted using a SPDE needle device, coated with a poly (dimethylsiloxane)(PDMS) phase. The detection was accomplished by GC-MS/MS in multiple reaction monitoring (MRM) mode. Parameters that affect the extraction ratio such as incubation time, extraction temperature, the number of extraction cycles, desorption volume and pH were also optimized. The optimized mass transition ion pairs (m/z) for quantitation were95.1/67.1for isoborneol and borneol,85.0/67.0for muscone,131.0/77.0for cinnamaldehyde, and128.1/102.1for IS. The method was thoroughly validated including specificity, linearity, precision, accuracy, recovery, and stability. The linear range were20-6250ng/mL for isoborneol,40~12500ng/mL for borneol,0.24-75ng/mL for muscone, and0.60~187.5ng/mL for cinnamaldehyde, respectively. The accuracy (RE%) of intra-day and inter-day ranged from-9.77.0%to9.87%, and the precisions (RSD%) were less than9.79%. The recoveries of all compounds were more than79.57%, and the stability of all the analytes with a concentration variation less than15.0%of the initial values. The method was applied to the analysis of plasma samples obtained after oral administration of SBP in rats. The pharmacokinetic parameters were calculated by WinNonlin5.2based on non-compartmental analysis of plasma concentration versus time data. The Gmax of isoborneol and borneol in rat plasma were3360.01±1432.56and5490.13±2136.47ng/mL, while the Tmax were1.00±0.52h and0.83±0.58h, respectively. And Cmax were23.54±4.33ng/mL for muscone and18.76±2.11ng/mL for cinnamaldehyde, which showed low plasma concentrations.In this study, the simple procedure for plasma sample preparation makes it promising for the analysis of complex volatile samples such as TCM (traditional Chinese medicine). In addition, the result acquired from this paper will be very helpful for the evaluation of the clinical applications of this formula and the research of compatibility mechanism.