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

Hao Chen,Yong-Jun Li,Yong-Lin Wang,Yong Huang,Lin Zheng,Zi-Peng Gong,Si-Ying Chen,Yan-Yu Lan,Ai-Min Wang,Chao-Ye Mei,Yue-Ting Li

doi:10.1155/2019/8942512

Hao Chen, Yong-Jun Li + Show 9 more

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https://doi.org/10.1155/2019/8942512

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Abstract

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 (R2 ≥ 0.999) over the concentration range with the lower limit of quantification between 0.015 and 0.029 μg/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 Tmax 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.

Highlights

Bletilla striata Reichb. f. (Orchidaceae) is a medicinal plant, the dry tubers of which have a hemostatic effect and are prescribed by the Chinese Pharmacopoeia as a medicinal part [1]
Our previous pharmacological work has confirmed that B. striata extract shows a hemostatic effect by promoting platelet aggregation [16, 17]
We found that α-isobutylmalic acid, gymnoside I, and militarine are relatively high in B. striata extract [19], and after oral B. striata extract in rats, three components were detected in the serum [20]

Summary

Introduction

Bletilla striata ( unb.) Reichb. f. (Orchidaceae) is a medicinal plant, the dry tubers of which have a hemostatic effect and are prescribed by the Chinese Pharmacopoeia as a medicinal part [1]. F. (Orchidaceae) is a medicinal plant, the dry tubers of which have a hemostatic effect and are prescribed by the Chinese Pharmacopoeia as a medicinal part [1] It is widely used in traditional Chinese herbal medicines for thousands of years in China and first recorded in Shennong’s Materia Medica [2]. It is bitter, sweet, and puckery in flavor and slightly cool and astringent in nature, and it acts on the lung, liver, and stomach meridians [1, 2]. Our previous pharmacological work has confirmed that B. striata extract shows a hemostatic effect by promoting platelet aggregation [16, 17]

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