chemical study on the rhizoma of curcuma longalibrary.um.edu.mo/etheses/b25903196_toc.pdf ·...

澳門大學碩士學位論文

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Chemical study on the rhizoma of Curcuma longa

by

Zhou Yanqing

Master of Science

2012

Institute of Chinese Medical Sciences

University of Macau


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Chemical study on the rhizoma of Curcuma longa

by

Zhou Yanqing

A thesis submitted in partial fulfillment of the

requirements for the degree of

Master of Science

Institute of Chinese Medical Sciences University of Macau

2012 Approved by __________________________________________________

Supervisor

__________________________________________________

__________________________________________________

__________________________________________________

Date __________________________________________________________


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In presenting this thesis in partial fulfillment of the requirements for a Master's

degree at the University of Macau, I agree that the Library and the Institute of

Chinese Medical Sciences shall make its copies freely available for inspection.

However, reproduction of this thesis for any purposes or by any means shall not

be allowed without my written permission. Authorization is sought by contacting

the author at

Address:

Telephone:

Fax:

E-mail:

Signature ______________________ Date __________________________


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碩士學位論文

薑黃的化學成分研究

研究生姓名：周燕青

導師：張慶文

專業：中藥學

日期： 2012 年 06 月

澳門大學中華醫藥研究院


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縮略詞表

縮寫英文全稱中文全稱

HPLC High-performance liquid chromatography 高效液相色譜

UV Ultra-Violet spectrum 紫外光譜

Prep-HPLC Preparative High Performance Liquid Chromatography 製備型高效液相色譜儀

HPLC-ESI-MS High-performance liquid chromatography-Electrospray Mass

Spectroscopy

高效液相色譜-電噴霧離子

阱質譜聯用技術

NMR Nuclear Magnetic Resonance 核磁共振

DEPT Distortionless Enhancement by Polarization Transfer

無畸變極化轉移增益

HMBC 1H-Detected Heteronuclear Multiple-Bond Correlation Spectroscopy

氫檢測異核多鍵相關譜

HSQC 1H-Detected Heteronuclear Single-Quantum

Coherence Spectroscopy

氫檢測異核單量子相關譜

1H-1H COSY Two Dimensional 1H Correlation Spectroscopy

1H-1H 同核相關譜

ROESY Rotating Frame Overhauser Effect Spectroscopy

旋轉坐標系中 NOE 效應

GC-MS Gas Chromatography- Mass spectrometry 氣相色譜-質譜聯用

HPCPC High Performance centrifugal partition chromatography 高效離心分配色譜

HSCCC high-speed countercurrent chromatography

高速逆流色譜

TLC Thin Layer Chromatography 薄層色譜


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摘要

薑黃為薑科薑黃屬(Curcuma)多年生草本植物薑黃(Curcuma longa L.)的乾燥根

莖，在亞洲的熱帶地區廣為栽培，廣泛用於食品添加劑及染料中。其味苦、辛，性

溫，歸脾、肝經，具有破血行氣、通經止痛之功效。現代藥理學研究，薑黃具有抗

炎、抗氧化、抗腫瘤、抗 HIV、抗老年癡呆以及抗微生物等多方面作用。其主要成

分為薑黃素類成分和揮發油成分。

為進一步探索薑黃的藥效成分，為其開發利用及品質控制提供科學依據，本論

文對薑黃進行了系統的化學成分研究。本論文分別對其乙醇提取物和石油醚提取物

進行了化學分離，運用矽膠柱層析色譜，葡聚糖凝膠柱色譜(Sephadex LH-20)，製備

型高效液相色譜(Prep-HPLC)，高速逆流色譜(HSCCC)等分離純化手段，分離得化合

物 20 個，並通過波譜解析和理化分析鑒定了它們的結構，分別為：薑黃素(CL-1)，

去甲氧基薑黃素 (CL-2) ，雙去甲氧基薑黃素 (CL-3) ，環薑黃素 (CL-4) ，

6-(4-hydroxy-3-methoxyphenethyl)-2-(4-hydroxy-3-methoxyphenyl)-2H-pyran-4(3H)-on

e(CL-5)，5-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)hept-1-en-3-one(CL-6)，六氫

薑黃素 (CL-7) ， 1,5-bis(4-hydroxy-3-methoxyphenyl)penta-1,4-dien-3-one(CL-8) ，

1-(4-hydroxy-3-methoxyphenyl)-5-(4-hydroxyphenyl)penta-1,4-dien-3-one(CL-9) ，

7-(4-hydroxy-3-methoxyphenyl)-1-(4-hydroxyphenyl)hepta-1,4,6-trien-3-one(CL-10) ，

4-(4-hydroxy-3-methoxyphenyl)but-3-en-2-one(CL-11), 芳薑黃酮(CL-12)，β-芳薑黃酮

(CL-13)，α-芳薑黃酮(CL-14)，E-α-大西洋酮(CL-15)，pyrocurzerenone (CL-16)，

bisabobolone(CL-17)，莪術二酮(CL-18)，4-hydroxybisabola-2,10-diene-9-one(CL-19)

和 β-穀甾醇(CL-20)，其中化合物 6 為新化合物。

在上述化學成分研究基礎上，本論文建立了 HPLC 方法同時對薑黃 3 個主要的

薑黃素類成份和 3 個主要挥发油成份進行含量測定，並對 26 批市售薑黃藥材進行了

定量分析。此方法簡單可靠，為首次報導採用 HPLC 法同時定量分析薑黃中薑黃素

类和揮發油這兩類物質，為薑黃品質控制研究提供重要的參考。

關鍵詞：薑黃；薑黃素类成份；揮發油；高速逆流色譜；含量分析；品質控制


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Abstract

Curcuma longa Linn.(Zingiberaceae) has been used in cosmetics, as a spice, food

preservative and coloring agent, and in medicinal preparations in Eastern civilization for

long time. In traditional chinese medicine, the rhizoma of C. longa named ‘JiangHuang’

has been widely used for the treatment of diabetic wounds, hepatic disorders and

cardiovascular disease. Previous phytochemical investigation indicated that the major

constituents of C. longa are curcuminoids and volatile oil. Studies of C. longa have

revealed numerous pharmacological properties, including anticancer, antioxidant,

anti-inflammatory, anti-parasitic, anti-HIV, antimicrobial and antiviral activities.

As part of a program to asses the chemical and biological diversities of medicinal

plants in south China, we had systematically carried out research on the chemical

constituents of C. longa. 20 compounds were isolated and purified by various column

chromatography on silica gel, Sephadex LH-20, Prep-HPLC, HSCCC and their structures

were identified by spectroscopic methods (MS, UV, 1D and 2D NMR) as follows:

curcumin(CL-1), demethoxycurcumin(CL-2), bisdemethoxycurcumin(CL-3),

cyclocurcumin(CL-4), 6-(4-hydroxy-3-methoxyphenethyl)-2-(4-hydroxy-3-methoxy

phenyl)-2H-pyran-4(3H)-one(CL-5), 5-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)

hept-1-en-3-one(CL-6), hexahydrocurcumin(CL-7), 1,5-bis(4-hydroxy-3-methoxy-

phenyl)penta-1,4-dien-3-one(CL-8), 1-(4-hydroxy-3-methoxyphenyl)-5-(4-hydroxy

phenyl)penta-1,4-dien-3-one(CL-9), 7-(4-hydroxy-3-methoxyphenyl)-1-(4-hydroxy

phenyl)hepta-1,4,6-trien-3-one(CL-10), 4-(4-hydroxy-3-methoxyphenyl)but-3-en-2-one

(CL-11), ar-Turmerone(CL-12), β-Turmerone(CL-13), α-Turmerone(CL-14),

E-α-atlantone (CL-15), pyrocurzerenone (CL-16), bisabobolone(CL-17), curdione

(CL-18), 4-hydroxybisabola-2,10-diene-9-one(CL-19) and β-sitosterol (CL-20),

repectively. Among them, compound 6 is a new compound.

In this thesis, an HPLC chromatogram method was also established for simultaneous

quantification of six major components in the rhizoma of C. longa, including three

curcuminoids and three volatile components. The validated method was applied to


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quantitatively analyze 6 major compounds in 26 ‘Jianghuang’ samples purchased from

different place in China, It is helpful for pharmacological evaluation and quality control

of C. longa.

Key words: Curcuma longa; curcuminoids; volatile oil; high-speed countercurrent

chromatography; quantification; Quality control


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本論文鑒定的化合物結構式

帶*號者為新化合物

CL-1: R1,R2=OCH3

CL-2: R1=H,R2=OCH3

CL-3: R1=H, R2=H

CL-4

CL-5

CL -6*

CL -7

CL -8

CL -9

CL -10

CL -11


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O

CL -12

CL -13

CL -14

CL -15

CL -16

CL -17

O

HO

H

CL -18

CL -19

CL -20


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目錄縮略詞表 ........................................................................................................................ I

摘要 ......................................................................................................................... III

Abstract ........................................................................................................................ V

本論文鑒定的化合物結構式 ................................................................................... VII

第一章薑黃的研究進展 ............................................................................................. 1 1 薑黃的化學成分 .......................................................................................................................... 1 2 提取與含量測定方法 .................................................................................................................. 5 3 藥理作用 ...................................................................................................................................... 6 4 結論 ............................................................................................................................................ 10

第二章薑黃的化學成份研究 ................................................................................... 21 第一節概述 ............................................................................................................................... 21 第二節化合物的結構鑒定 ......................................................................................................... 22 第三節實驗部分 ......................................................................................................................... 39 第四節逆流色譜法分離薑黃油主要成份 ................................................................................. 49

第三章薑黃主要成分含量測定 ............................................................................... 63 1 儀器和材料 ............................................................................................................................... 63 2 方法和結果 ............................................................................................................................... 64 3 討論 ........................................................................................................................................... 69 4 結論 ........................................................................................................................................... 73

附錄目錄 ...................................................................................................................... 77

附錄 ............................................................................................................................ 81

致謝 .......................................................................................................................... 121

個人簡介 .................................................................................................................... 123

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