液相層析串聯質譜法鑑別生曬參、紅參、西洋參、枳殼、黄耆及紅耆中的指標成分

Development of LC/MS Methods for Identification of Marker Components in Panax ginseng, Panax quinquefolinm, Citrus aurantium, Astragalus membranaceus and Hedysarum polybotrys

陳威廷(Wei-Ting Chen)

指導教授 ： 王賢達

國立臺北科技大學/工程學院/有機高分子研究所/碩士(2005年)

摘要

在本研究中，我們開發了三種液相層析串聯質譜（Liquid Chromatography/ Mass Spectrometry，LC/MS）方法來辨認藥材中的化學指標成分，分別針對生曬參、紅參及西洋參中的六個人參皂苷（Ginsenosides Rb1、Ginsenosides Rg1、Ginsenosides Rc、Ginsenosides Rg3、Ginsenosides Re和Ginsenosides Rh2），枳殼中的七個黃酮類化合物（Synephrine、Narirutin、Naringin、Hesperidin、Quercertin、Naringenin 和Imperatorin）與黃耆及紅耆中的七個黃酮類化合物（Calycosin-7-O-β-D-glucoside、Calycosin-7-O-β-D-glucoside -6-O-malonate、Ononin、Calycosin、Formononetin-7 -O-β-D -glucoside-6-O-malonate、Astragaloside IV和 Formononetin）進行分析鑑定。
此外，我們還利用LC/MS/MS對黃耆中的二個黃酮類化合物（Astragaloside IV和Formononetin）進行二次質譜的鑑定。
液相層析部分，靜相是使用Cosmosil ARII C18 5μm 2.1×150mm的管柱，移動相是使用醋酸緩衝液及Acetonitrile進行梯度沖提，而偵測波長分別為210nm、254nm和280nm。
串聯質譜部分，離子化方式是以電噴灑法（Electrospray Ionization，ESI），質量分析儀採用四極柱（Quadrupole）和飛行時間（Time of Flight）組合，帶動氣體（Carrier gas）使用氮氣，而碰撞氣體（Collision gas）使用氬氣。
並使用對照成份以進行辨認生曬參、紅參、西洋參、枳殼、黄耆及紅耆，於紫外光光譜與質譜中的化學成分，並製作藥材的化學指紋圖譜，利用這些化學指紋圖譜，可區別這些藥材的基原品種。

關鍵字

液相層析串聯質譜法 ； 生曬參 ； 紅參 ； 西洋參 ； 枳殼 ； 黃耆 ； 紅耆

並列摘要

Liquid chromatography/mass spectrometry ( LC/MS ) methods were developed to identify six ginsenosides ( Ginsenosides Rb1, Ginsenosides Rg1, Ginsenosides Rc, Ginsenosides Rg3, Ginsenosides Re and Ginsenosides Rh2 ) in Panax ginseng and Panax quinquefolinm , seven flavonoids ( Synephrine, Narirutin, Naringin, Hesperidin, Quercertin, Naringenin and Imperatorin ) in Citrus aurantium and seven flavonoids ( Calycosin-7-O-β-D-glucoside, Calycosin-7-O-β-D-glucoside -6-O-malonate, Ononin, Calycosin, Formononetin-7-O-β-D-glucoside-6-O-malonate, Astragaloside IV and Formononetin ) in Astragalus membranaceus and Hedysarum polybotrys.
LC was carried out using a C18 column with linear gradient elution of acetate buffer- acetonitrile and detected at the ultraviolet (UV) wavelength of 210nm, 254nm and 280nm, separately.
The tandem MS was used with positive-ion electrospray ionization, Quadrupole and Time of Flight (TOF) analyzers, and argon as a collision gas.
Ginsenosides and flavonoids mentioned above were identified on both LC/UV and LC/MS spectra of these studied herbs compared with standards.
These marker components in Panax ginseng, Panax quinquefolinm, Citrus aurantium, Astragalus membranaceus and Hedysarum polybotrys have been assembled as fingerprints in LC/UV or LC/MS spectra of identifying Chinese herbs.

並列關鍵字

Panax ginseng ； Panax quinquefolinm ； Citrus aurantium ； Astragalus membranaceus ； Hedysarum polybotrys ； LC/MS ； Flavonoids ； Ginsenosides ； Chinese herb

參考文獻

[50] 陳月枝，現代生化質譜術-基質輔助雷射脫附游離質譜法，Chemistry，No.2，2002，pp.229 ~ 240.

[3] Chang-Hwa Jung, Ho-Moon Seog , In-Wook Choi , Hee-Don Choi , Hong-Yon Cho, Effects of wild ginseng (Panax ginseng C.A. Meyer) leaves on lipid peroxidation levels and antioxidant enzyme activities in streptozotocin diabetic rats, Journal of Ethnopharmacology , 98, 2005, pp.245~250.

[4] Khaled Radad, Gabriele Gille, Rudolf Moldzio, Hiroshi Saito, Wolf-Dieter Rausch, Ginsenosides Rb1 and Rg1 effects on mesencephalic dopaminergic cells stressed with glutamate., Brain Research ,1021 , 2004 , pp. 41~ 53.

[6] Yoshiki Y., Kudou S., Okubo K., Relationship Between Chemical Structures and Biological Activities of Triterpenoid Saponins from Soybean., Biosci. Biotechnol. Biochem. , 62, 1998, pp.2291~2300.

[7] Akiyama T., Tanaka O., Shibata S.,Chemical Studies on the Oriental Plant Drugs, XXXII. Saponins of the Roots of Platycodon Grandiflorum A. De Candolle(3). The Structure of a Prosapogenin, 3-O-B-Glucosylplatycodigenin., Chem. Pharm.Bull., 20(9), 1972, pp.1957~1961.