运用HPLC-MS对鸦胆子油自微乳给药系统肠吸收的研究

High-performance Liquid Chromatography Coupled with Mass Spectrometric Method for the Intestinal Absorption Study of Brucea Javanica Oil SMEDDS

目的：建立高效液相色谱-串联质谱法检测油酸和亚油酸含量的方法，从而对鸦胆子油自微乳给药系统中鸦胆子油的肠吸收进行研究。方法：以甲醇．水（95：5v／v）为流动相，流速为0．4mL/min，柱温为35℃作为高效液相色谱的检测条件。利用大鼠小肠膜建立体外药物扩散体系研究鸦胆子油的肠吸收特性。结果：油酸和亚油酸的保留时间分别为10．46t0．02和8．55±0．01min，线性范围分别为0．50±50．0ng／mL和5．06±101．2ng／mL，平均绝对回收率分别为97．49±3．11％和105．76±3．13％。日间和日内精密度都小于5％。在肠吸收实验中，鸦胆子油自微乳体系中测得油酸和亚油酸含量是单独给予鸦胆子油测得量的2．8和4．1倍。结论：该方法高效、灵敏、选择性高，可以用于鸦胆子油及鸦胆子油自微乳肠吸收中油酸和亚油酸的质量测定。

ABSTRACT Objective： To develop a rapid, sensitive and selective high-performance liquid chromatography coupled with mass spectrometric method （HPLC-MS） for detection oleic acid and linoleic acid. Methods： The chromatographic separation was achieved on C18 column at 35 ℃, with a mobile phase consisting of methanol-dlstiUed water （95：5, v/v） at a flow rate of 0A mL/min. An in-vitro diffusion chamber system across isolated rat intestinal membranes was chosen as a model. A self-microemulsifying drug delivery system was used to enlaance the intestinal absorption of brutes javanica oil. Results： Oleicacid and linoleic acid were separated with retention times of 10.46 ± 0.02 and 8.55 ± 0.01 min, respectively. A good linear relationship for oleic acid and linoleic acid were in the range of 0.50±50.0 ng/mL（oleic acid） and 5.06-101.2 ng/mL（linoleic acid）, respectively. The mean absolute recoveries of oleic acid and linoleic acid determined in middle concentrations were 97.49± 3.11% and 105.76± 3.13 % respectively. The coefficients of variation for inter-day and intra-day assay were less than 5 %. The absorption of oleic acid and linoleic acid in brutes javanica oil were 2.8-fold and 4.1-fold enhancement in the presence of the self-mieroemulsifying drug delivery system respectively, compared with bmcea javanica oil alone. Conclusion： HPLC-MS method will be of great utility in routine quality control procedure for the determination of oleic acid and linoleic acid in absorption experiments. Key words： Brucea javanica oil; Oleinic acid; Linoleic acid; HPLC-MS; Self-microemulsifying drug delivery system