Objective: To establish a supercritical fluid chromatography method for the simultaneous determination of 7 coumarins (oxypeucedanin hydrat, bergapten, oxypeucedanin, imperatorin, osthole, isoimperatorin, suberosin) in Angelica dahurica, and to compare with high performance liquid chromatography(HPLC) method. Methods: The Agilent ZORBAX RX-SIL (150 mm×4.6 mm, 5 μm) column was used, the mobile phase was CO2(A)-ethanol(B) with gradient elution, flow rate was 1.5 mL·min-1, the column temperature was 45 ℃, back pressure was 12.75 MPa, the injection volume was 2 μL, and the detection wavelength was 280 nm. Results: The seven coumarins all had good linear relationship(r>0.999 0), the RSDs of precision were 0.79%-2.9%, seven coumarins remained stable within 24 h, the RSDs of repeatability were 1.8%-3.0%, and the recoveries were 93.8%-102%. In 27 batches of Angelica dahurica decoction piece, seven of coumarin compounds content from high to low were imperatorin, oxypeucedanin hydrat, isoimperatorin, oxypeucedanin, bergapten, suberosin, and osthole, the contents ranged from 1.00 to 5.70, 0.24 to 3.10, 0.31 to 1.50, 0.01 to 2.50, 0.03 to 0.39, 0.01 to 1.20, 0.01 to 0.54 mg·g-1, respectively. To some extent, the SFC method and the HPLC method showed the opposite chromatographic behavior. Conclusion: The established SFC method can rapidly separate 7 coumarins including 2 pairs of isomers. The retention and selectivity of SFC method are greatly affected by column and modifier. Compared with HPLC method, the SFC method is more rapid, economic and environmental friendly.
[1] 中华人民共和国药典 2020年版. 一部[S].2020: 109
ChP 2020. Vol Ⅰ[S].2020: 109
[2] ZHAO AH, ZHANG YB, YANG XW. Simultaneous determination and pharmacokinetics of sixteen Angelicae dahurica coumarins in vivo by LC-ESI-MS/MS following oral delivery in rats[J].Phytomedicine, 2016, 23(10):1029
[3] YANG YF, ZHANG L, YANG XW. Distribution assessments of coumarins from Angelicae Pubescentis Radix in rat cerebrospinal fluid and brain by liquid chromatography tandem mass spectrometry analysis[J].Molecules, 2018, 23(1):225
[4] SINGH G, SINGH A, SINGH P, et al. Bergapten ameliorates vincristine-induced peripheral neuropathy by inhibition of inflammatory cytokines and NFκB signaling[J].ACS Chem Neurosci, 2019, 10(6):3008
[5] HE Y, ZENG ZS, LU ZH, et al. Bergapten alleviates osteoarthritis by regulating the ANP32A/ATM signaling pathway[J].FEBS Open Bio, 2019, 9(6):1144
[6] SINGH G, KAUR A, KAUR J, et al. Bergapten inhibits chemically induced nociceptive behavior and inflammation in mice by decreasing the expression of spinal PARP, iNOS, COX-2 and inflammatory cytokines[J].Inflammopharmacology, 2019, 27(4):749
[7] KANG TJ, LEE SY, SINGH RP, et al. Anti-tumor activity of oxypeucedanin from Ostericum koreanum against human prostate carcinoma DU145 cells[J].Acta Oncol, 2009, 48(6):895
[8] YANG HB, GAO HR, REN YJ, et al. Effects of isoimperatorin on proliferation and apoptosis of human gastric carcinoma cells[J].Oncol Lett, 2018, 15(5):7993
[9] 王笃军, 王硕, 严峰, 等. 欧前胡素和异欧前胡素对小鼠肝CYP450的影响[J].中成药, 2017, 39(1):14
WANG DJ, WANG S, YAN F, et al. Effects of imperatorin and isoimperatorin on liver cytochrome P450 in mice[J].Chin Tradit Pat Med, 2017, 39(1):14
[10] YANG W, YE M, LIU M, et al. A practical strategy for the characterization of coumarins in Radix Glehniae by liquid chromatography coupled with triple quadrupole-linear ion trap mass spectrometry[J].J Chromatogr A, 2010, 1217(27):4587
[11] 邓瑞, 张静, 罗维早, 等. RRLC-UV同时测定川白芷中6种香豆素类成分的含量[J].中国药学杂志, 2010, 35(23):3184
DENG R, ZHANG J, LUO WZ, et al. Quantitative determination of six coumarins in Angelica dahurica var. formosana by RRLC-UV[J].Chin Pharm J, 2010, 35(23):3184
[12] 曾承辉, 王晓可, 毋福海. 毛细管电泳法测定白芷药材中欧前胡素和异欧前胡素的含量[J].中药材, 2009, 32(12):1850
ZENG CH, WANG XK, WU FH. Determination of imperatorin and isoimperatorin in Angelica dahurica by capillary electrophoresis[J].J Chin Med Mater, 2009, 32(12):1850
[13] 席梅, 王欣, 陆艳. 白芷有效成分含量测定及其指纹图谱研究[J].世界中医药, 2016, 11(11):2434
XI M, WANG X, LU Y. Determination of effective ingredient and fingerprint of Dahurian Angelica Radix[J].World Chin Med, 2016, 11(11):2434
[14] 张烨, 孙艳雪, 马宇衡, 等. 白芷HPLC指纹图谱的研究[J].食品与药品, 2020, 22(2):130
ZHANG Y, SUN YX, MA YH, et al. Study on HPLC fingerprints of Angelicae Dahuricae Radix[J].Food Drug, 2020, 22(2):130
[15] 中华人民共和国药典 2020年版. 四部[S].2020: 68
ChP 2020. Vol Ⅳ[S].2020: 68
[16] KALIKOVA K, SLECHTOVA T, VOZKA J, et al. Supercritical fluid chromatography as a tool for enantioselective separation: a review[J].Anal Chim Acta, 2014, 821: 1
[17] LAURENT L, MARIO O, DAVID T. Lipidomics by supercritical fluid chromatography[J].Int J Mol Sci, 2015, 16(6):13868
[18] 陈琳, 唐志书, 宋忠兴, 等. 不同产地白芷药材9个呋喃香豆素成分的含量测定及其质量评价[J].中国中药杂志, 2019, 44(14):3002
CHEN L, TANG ZS, SONG ZX, et al. Quantitative determination of nine furanocoumarins for quality evaluation of Angelica dahurica from different habitats[J].China J Chin Mater Med, 2019, 44(14):3002
[19] 蒋运斌, 卢晓琳, 杨枝中, 等. 与熏硫加工相关的白芷化学成分研究[J].中国实验方剂学杂志, 2013, 19(22):74
JIANG YB, LU XL, YANG ZZ, et al. Studies on the chemical constituents of Angelica dahurica related to sulfur fumigated[J].Chin J Exp Tradit Med Form, 2013, 19(22):74
[20] LIN P, WANG Q, LIU Y, et al. Characterization of chemical profile and quantification of representative components of Dan Lou tablet, a traditional Chinese medicine prescription by UHPLC-Q/TOF-MS combined with UHPLC-TQF-MS-Science direct[J].J Pharm Biom Anal, 2020, 180: 113070
[21] WEST C, LESELLIER E. A unified classification of stationary phases for packed column supercritical fluid chromatography[J].J Chromatogr A, 2008, 1191(1-2):21
[22] LUNDGREB J, SALOMONSSON J, GYLLENHAAL O. Supercritical fluid chromatography of metoprolol and analogues on aminopropyl and ethylpyridine silica without any additives[J].J Chromatogr A, 2007, 1154(1-2):360
[23] GRAND-GUILLAUME PERRENOUD A, BOCCARD J, VEUTHEY JL, et al. Analysis of basic compounds by supercritical fluid chromatography: attempts to improve peak shape and maintain mass spectrometry compatibility[J].J Chromatogr A, 2012, 1262: 205
[24] JIANG C, REN Q, WU P. Study on retention factor and resolution of tocopherols by supercritical fluid chromatography[J].J Chromatogr A, 2003, 1005(1-2):155
[25] JONES MD, AVULA B, WANG YH, et al. Investigating sub-2 μm particle stationary phase supercritical fluid chromatography coupled to mass spectrometry for chemical profiling of chamomile extracts[J].Anal Chim Acta, 2014, 847: 61
