臺灣博碩士論文加值系統

本研究是開發以液相層析串聯質譜術在電灑游離法正離子模式下偵測半夏與兩種炮製半夏及七種含半夏之中藥複方中生物鹼成分Ephedrine、Harman、Norharman、Harmine之定量分析之研究。在本篇研究中，成功地發展出快速具選擇性的LC/ESI/MS/MS的方法，對半夏中四個生物鹼類成分做定量分析之研究。定量結果顯示偵測極限(LODs)分別為Ephedrine：4.5 ng/g；Harman：2.5 ng/g；Norharman：1.5 ng/g；Harmine：4.2 ng/g，並且Ephedrine訊號強度在10 - 250 ng/g以及Harman、Norharman、Harmine訊號強度在5 - 250 ng/g之濃度範圍內呈現良好的線性，線性相關係數皆在0.995以上。同日間(Intra–day)精密度相對標準偏差在4.4 - 18.7 % 之間，異日間(Inter–day)精密度相對標準偏差在7.3 - 19.5% 之間，顯示具良好定量效能之精密度。回收率對於半夏複方(半夏瀉心湯)萃取方法在92 % - 105 %之間，顯示具良好之回收率 。
在生半夏與兩種半夏單方(姜半夏、法半夏)中，實驗結果顯示Ephedrine、Harman、Norharman與Harmine生物鹼總含量由多至少為生半夏>姜半夏>法半夏，Ephedrine含量在7.3 - 70.3 ng/g之間；Harman含量在1.1 - 18.8 ng/g之；Norharman含量在3.2 - 10.0 ng/g之間；Harmi ne含量在N.D. - 8.3 ng/g之間。
在七種含半夏之中藥複方，實驗結果顯示Ephedrine含量在6.7 - 37.2 ng/g之間；Harman含量在5.8 – 31.3 ng/g之間；Norharman含量在10.6 – 48.6 ng/g之間；Harmine含量在N.D. – 6.5 ng/g之間。
由炮製半夏法半夏定量結果，可將三種β–carbolines alkaloids總含量回朔至各科學中藥複方中半夏之含量百分比。

Determination of four marker compounds ephedrine, harman, norharman and harmine were carried out in crude methanol extractions of Pinellia tubers derived from three different processing methods and of seven most popular Chinese medicinal preparations containing Pinellia tubers as the ingredient without any clean-up by using a HPLC / tandem Mass analysis. The best ionization characteristics were obtained using positive-ion electrospray ionization (ESI) and 0.05 % formic acid as additive. Good linearity over the range 10 - 250 ng/g for ephedrine and range 5 - 250 ng/g for harman, norharman and harmine were observed. The intra and inter precision within quantitation ranges varied between 4.4 - 18.7 % and 7.3 - 19.5 % respectively. The limit of detection was 4.5, 2.5, 1.5 and 4.2 ng/g for ephedrine , harman , norharman and harmine respectively. Recoveries from extract of the medicinal preparation sample Bann-Shiah-Shieh-Shin-Tang were 92, 96, 103 and 105 % respectively at concentration of 100 ng/mL spiking of harman , norharman , ephedrine and harmine. The amount of total alkloids vary in these Pinellia products in the following order: raw Pinellia > Pinellia dipped in ginger and alum juice > Pinellia dipped in Glycyrrhizae Radix plus sodium carbonate solution.
The determination of the four markers in seven Chinese medicinal preparations was ranged between 5.8 – 31.3 ng/g for harman , 10.6 – 48.6 ng/g for norharman , 6.7 - 37.2 ng/g for ephedrine and N.D. – 6.5 ng/g for harmine.

摘要................................................................................................................i Ab-stract........................................................................................................iii目次...............................................................................................................v
表次............................................................................................................viii
圖次...............................................................................................................x
第一章 前言.................................................................................................1
1.1半夏藥材之簡介.....................................................................................1
1.2 半夏相關複方藥材之簡介....................................................................3
1.3 文獻回顧................................................................................................5
1.4 半夏藥理活性........................................................................................6
1.5 研究動機................................................................................................7
第二章 實驗與分析方法...........................................................................10
2.1 實驗藥品與藥材..................................................................................10
2.2 標準樣品之配製..................................................................................13
2.3 校正曲線之配製..................................................................................13
2.4真實樣品的製備與萃取.......................................................................14
2.4.1 半夏單方萃取...................................................................................14
2.4.2 半夏複方萃取...................................................................................14
2.5 回收率實驗..........................................................................................15
2.6 液相層析條件......................................................................................15
2.7 最佳化之質譜條件..............................................................................16
第三章 結果與討論...................................................................................18
3.1 生物鹼標準品之液相層析質譜術分析..............................................18
3.1.1 Ephedrine之質譜術分析..................................................................18
3.1.2 Norharman之質譜術分析................................................................18
3.1.3 Harman之質譜術分析.....................................................................19
3.1.4 Harmine之質譜術分析....................................................................19
3.2 內標準品的選擇與質譜術分析..........................................................19
3.3 質譜參數最佳化..................................................................................27
3.4 動相添加劑的影響..............................................................................31
3.5 選擇反應偵測(SRM)的質譜層析圖...................................................33
3.6分析物之校正曲線、線性範圍及偵測極限.......................................37
3.7 LC/ESI(+)/MS/MS方法分析半夏中藥材中生物鹼成分(Ephedrine、
Norharman、Harman 及 Harmine)之定量效能精密度....................41
3.8 LC/ESI(+)/MS/MS方法分析半夏中藥材中生物鹼成分(Ephedrine、Norharman、Harman 及 Harmine)之複方萃取回收率.....................43
3.9 真實樣品分析......................................................................................45
第四章 結論...............................................................................................50
參考文獻.....................................................................................................51

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