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A fast, sensitive, and high‐throughput method for the simultaneous quantitation of three ellagitannins from Euphorbiae pekinensis R adix in rat plasma by ultra‐ HPLC – MS / MS
Author(s) -
Hou Pengyi,
Zeng Yan,
Ma Bingjie,
Wang Xiaofan,
Liu Zhenzhen,
Li Lu,
Qu Kankan,
Bi Kaishun,
Chen Xiaohui
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300389
Subject(s) - chemistry , chromatography , ellagic acid , extraction (chemistry) , high performance liquid chromatography , analyte , selected reaction monitoring , ethyl acetate , mass spectrometry , tandem mass spectrometry , biochemistry , polyphenol , antioxidant
A fast, sensitive, and high‐throughput ultra‐ HPLC – MS / MS method has been developed and validated for the simultaneous determination of three main active constituents of E uphorbiae pekinensis R adix in rat plasma. After addition of the internal standard, plasma samples were extracted by liquid–liquid extraction with ethyl acetate/isopropanol (1:1, v/v) and separated on a CAPCELL PAK C 18 column (100 × 2.0 mm, 2 μm, S hiseido, J apan), using a gradient mobile phase system of methanol/water. The detection of the analytes was performed on a 4000 Q UHPLC – MS / MS system with turbo ion spray source in the negative ion and multiple reaction‐monitoring mode. The linear range was 1.0–1000 ng/mL for 3,3′‐di‐ O ‐methyl ellagic acid‐4′‐ O ‐ β ‐d ‐glucopyranoside (i), 1.5–1500 ng/mL for 3,3′‐di‐ O ‐methyl ellagic acid‐4′‐ O ‐ β ‐d ‐xylopyranoside (ii), and 5.0–5000 ng/mL for 3,3′‐di‐ O ‐methyl ellagic acid (iii). The intra‐ and interday precision and accuracy of all the analytes were within 15%. The extraction recoveries of the three analytes and internal standard from plasma were all more than 80%. The validated method was first successfully applied to the evaluation of pharmacokinetic parameters of compounds 1 , 2 , and 3 in rat plasma after intragastric administration of the E uphorbiae pekinensis R adix extract.