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Simultaneous determination of seven major diterpenoids in S iegesbeckia pubescens M akino by high‐performance liquid chromatography coupled with evaporative light scattering detection
Author(s) -
Huo Linan,
Jiang Zhen,
Li Huining,
Wang Mingsheng,
Ye Xiaoxia,
Ji Bin,
Guo Xingjie
Publication year - 2012
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.201200286
Subject(s) - chromatography , chemistry , formic acid , chromatography detector , high performance liquid chromatography , calibration curve , analytical chemistry (journal) , detection limit
A novel HPLC method with evaporative light scattering detection was developed for the simultaneous quantification of seven major diterpenoids of two types, including ent ‐pimarane type: K irenol, H ythiemoside B , D arutigenol, and ent ‐kaurane type: ent ‐16β,17,18‐trihydroxy‐kauran‐19‐oic acid, ent ‐17,18‐dihydroxy‐kauran‐19‐oic acid, ent ‐16β,17‐dihydroxy‐kauran‐19‐oic acid, 16α‐hydro‐ ent ‐kauran‐17,19‐dioic acid in the aerial parts of S iegesbeckia pubescens M akino, an important traditional C hinese medicinal herb. Chromatographic separation was achieved on a W aters S ymmetry S hield TM RP 18 column (250 mm× 4.6 mm id, 5 μm) with a gradient mobile phase (A: 0.3% v/v aqueous formic acid and B: acetonitrile) at a flow rate of 1.0 mL/min. The drift tube temperature of evaporative light scattering detection was set at 103°C, and nitrogen flow rate was 3.0 L/min. The method was validated for accuracy, precision, LOD , and LOQ . All calibration curves showed a good linear relationship ( r > 0.999) in test range. Precision was evaluated by intra‐ and interday tests that showed RSD s were less than 3.5%. Accuracy validation showed that the recovery was between 96.5 and 102.0% with RSD s below 2.8%. The validated method was successfully applied to determine the contents of seven diterpenoids in the different parts of S iegesbeckia pubescens M akino from two sources and to determine the contents of ent ‐pimarane, ent ‐kaurane, and total diterpenoids.