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Bletilla striata  has been widely used as a valuable hemostatic agent for thousands of years due to the high levels of bioactive constituents it contains. Here, we used a sensitive ultrahigh-performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) method for the simultaneous determination of three major active ingredients of the  B. striata  extract, namely,  α -isobutylmalic acid, gymnoside I, and militarine in rat plasma. The three major active ingredients were determined using the multiple reaction monitoring (MRM) mode at  m/z  189 ⟶ 129 for  α -isobutylmalic acid,  m/z  457.2 ⟶ 285.1 for gymnoside I,  m/z  725.3 ⟶ 457.2 for militarine, and  m/z  417.0 ⟶ 267.0 for the IS puerarin. All calibration curves showed good linearity ( R  2  ≥ 0.999) over the concentration range with the lower limit of quantification between 0.015 and 0.029  μ g/mL. The relative standard deviations of intraday and interday measurements were less than 15%, and the method was accurate within 93.3–100.4%. The extraction recovery was 92.65–100.98%, and no matrix effect was observed. The results indicated that after oral administration of  B. striata  in rats, the  T  max  of  α -isobutylmalic acid was significantly longer than that of gymnoside I and militarine and the mean residence time and area under the curve of  α -isobutylmalic acid and gymnoside I in rats were significantly higher than those of militarine. Moreover, the blood concentration-time curve of  α -isobutylmalic acid showed double peaks, suggesting that  α -isobutylmalic acid could exhibit the phenomenon of enterohepatic circulation or metabolic conversion. We also explored some of the pharmacokinetic characteristics of three ingredients from  B. striata  extract  in vivo , and the data obtained may provide a basis for the further investigation of  B. striata .

evidence-based complementary and alternative medicine

Simultaneous Determination of Three Bioactive Constituents from <i>Bletilla striata</i> by UPLC-MS/MS and Application of the Technique to Pharmacokinetic Analyses

Simultaneous Determination of Three Bioactive Constituents from Bletilla striata by UPLC-MS/MS and Application of the Technique to Pharmacokinetic Analyses