高效液相色谱法拆分阿雷地平对映异构体

目的：建立阿雷地平对映异构体高效液相色谱手性拆分方法，用于S构型阿雷地平光学纯度的分析测定。方法：采用硅胶表面涂敷有纤维素-三（4-甲基苯甲酸酯）填料（Chiralcel OJ-H）为固定相，以正己烷-乙醇（75∶25）为流动相，流速1.0 mL·min^-1，检测波长238 nm，柱温35℃。结果： S-阿雷地平与R-阿雷地平之间的分离度为3.5；R-阿雷地平质量浓度在0.04~2.0 μg·mL^-1范围内线性关系良好，线性方程为A=61 971C-0.321 4，r=0.999 9；R-阿雷地平的定量限和检测限分别为0.8 ng和0.3 ng；低、中、高3个浓度的R-阿雷地平回收率（n=3）分别为97.5%、98.5%、99.8%，RSD（n=9）为1.5%；供试品溶液在避光条件下室温放置12 h内稳定性良好。经检测，S-阿雷地平中R-阿雷地平的含量为0.08%。结论：所建立的方法经方法验证可用于分离和测定S-阿雷地平中的R-阿雷地平。

Objective: To establish an HPLC method to separate aranidipine enantiomers for optical purity detection of S-aranidipine.Methods: The method was performed on a Chiralcel OJ-H column[cellulose tri (4-methylbenzoate)chiral stationary phase].The mobile phase was n-hexane-ethanol(75:25)at a flow rate of 1.0 mL·min^-1.The column temperature was 35℃ and the detection wavelength was 238 nm.Results: The resolution between the peaks of S-aranidipine and R-aranidipine was 3.5.The method was found to show good linearity over the concentration range of 0.04-2.0 μg·mL^-1.The linear equation was A=61 971C-0.321 4, and the regression coefficient was 0.999 9.The limit of quantification and limit of detection of R-aranidipine were 0.8 ng and 0.3 ng, respectively.The recoveries of R-aranidipine(n=3)were 97.5%,98.5%,and 99.8% respectively at the concentration level of low,medium and high,with the RSD(n=9)of 1.5%.The test solution was stable for at least 12 h at 25℃ away from light.The R-aranidipine content in S-aranidipine sample was 0.08%.Conclusion: With good resolution and repeatability, the method can be reliably used for chiral separation of aranidipine, and for determination of the optical purity of S-aranidipine.

[1] OHNO S, KOMATSU O, MIZUKOSH IK, et al. Synthesis of asymmetric 4-aryl-1, 4-dihydro-2, 6-dimethyl-3, 5-pyridinedicar-boxylates with vasodilating and antihypertensive activities[J]. Chem Pharm Bull, 1986, 34(4):1589
[2] KANDA A,HARUNO A,MIYAKE H,et al. Antihypertensive effects of MPC-1304,a novel calcium antagonist,in experimental hypertensive rats and dogs[J]. J Cardiovasc Pharm,1992,20(5):723
[3] MIYOSHI K, KANDA A, MIYAKE H, et al. MPC-1304, another type of dihydropyridine,possessing highly potent vasodilating action[J]. Eur J Pharmacol, 1993, 238(2):139
[4] MASUMIYA H,TANAKA Y,TANAKA H,et al. Inhibition of T-type and L-type Ca²⁺ currents by aranidipine,a novel dihydropyridine Ca²⁺ antagonist[J]. Pharmacology,2000,61(2):57
[5] IOAN P, CAROSATI E, MICUCCI M, et al. 1, 4-Dihydropyridine scaffold in medicinal chemistry, the story so far and perspectives (part 1):action in ion channels and GPCRs[J]. Curr Med Chem, 2011, 18(32):4901
[6] WANG S,HE L,YUN B. The difference between nicardipine and its enantiomers on inhibiting vasoconstriction of isolated rabbit thoracic artery[J]. Arch Pharm Res, 2005,28(3):319
[7] JABOR VA, COELHO EB, LANCHOTE VL. Enantioselective pharmacokinetics of lercanidipine in healthy volunteers[J]. J Chromatogr B, 2004, 813(1):343
[8] ZHANG XP, LOKE KE, MITAL S, et al. Paradoxical release of nitric oxide by an L-type calcium channel antagonist, the R+ enantiomer of amlodipine[J]. J Cardiovasc Pharmacol, 2002, 39(2):208
[9] OKUMURA K, ICHIHARA K, NAGASAKA M, et al. Calcium entry blocking activities of MPC-1304 and of its enantiomers and metabolites[J]. Eur J Pharmacol, 1993, 235(1):69
[10] 郭鹏程,黄红林.二氢吡啶类钙通道阻滞剂及其药物手性研究[J].现代生物医学进展, 2009, 9(1):181 GUO PC, HUANG HL. Dihydropyridines calcium channel blockers and chiral drugs research[J]. Prog Mod Biomed, 2009, 9(1):181
[11] TIAN L, JIANG J, HUANG Y, et al. Determination of aranidipine and its active metabolite in human plasma by liquid chromatography/negative electrospray ionization tandem mass spectrometry[J]. Rapid Commun Mass Spectrom, 2006, 20(19):2871
[12] ⅡDA Y, KINOUCHI Y, TAKEICHI Y, et al. Simultaneous determination of a new dihydropyridine calcium antagonist(MPC-1304)and its metabolite in dog plasma by high-performance liquid chromatography with electrochemical detection[J]. J Chromatogr A, 1991, 571(1-2):277
[13] 李成平,曾怀超,鲁琳,等.二氢吡啶钙拮抗剂马尼地平和西尼地平对映体的手性拆分研究[J].药物分析杂志,2010,30(4):661 LI CP, ZENG HC, LU L, et al. Study on the chiral resolution of dihydropyridine calcium antagonistsmanidipine and cilnidipine enantiomers[J]. Chin J Pharm Anal, 2010, 30(4):661
[14] 方应国,丁潇潇,严小平,等.非洛地平和尼莫地平对映体的手性拆分研究[J].药物分析杂志, 2012,32(10):1762 FANG YG, DING XX, YAN XP, et al. Study on chiral resolution of felodipine and nimodipine enantiomers[J]. Chin J Pharm Anal, 2012, 32(10):1762
[15] 赵蕊.阿雷地平的合成研究[D].天津:天津理工大学,2011 ZHAO R. Synthesis of Aranidipine[D]. Tianjin:Tianjin University of Technology, 2011