高效液相層析（HPLC）和毛細管電泳( CE )同屬液相分離技術，具有多種分離模式，為目前最常用於定量中藥材指標成分的分析方法，本研究應用這兩種儀器於厚朴、吳茱萸藥材的分析。LC-UV及CE-UV的偵測範圍有一定的極限，對藥材微量成分或動物血液組成的分析，常有困難，本文開發毛細管電泳掃集法，成功地測定小白鼠血液中麻黃生物鹼的含量及厚朴藥材中微量生物鹼的含量。
麻黃的主要藥效成分為六個麻黃生物鹼，1-ephedrine (E)、d-pseudo-
ephedrine (PE)、1-norephedrine (NE)、d-norpesudoephedrine (NPE)、1-methylephedrine (ME) 和d -methylpseudoephedrine (MPE)，利用場放大電動注射掃集法（CSEI-Sweep）分析，偵測極限可達20.0～70.3ppb；本分析方法以50 mM SDS之25 mM 磷酸溶液，及氰甲烷為緩衝溶液( 80：20,v/v )，在27分鐘內能完全分離出六成分峰，再現性良好，RSD值在1.06～2.86 ﹪之間。該方法並可應用於小白鼠的血液分析，測定極稀溶液中組成成分的存在及含量。
厚朴為常用中藥材，具溫中下氣、燥濕消痰之效，含有酚類magnolol（M）、honokiol（H），及微量生物鹼magnocurarine（A1）、salicifoline（A2）、magnoflorine、anonaine、michelarbine、liriodenine及laurifoline等成分，其中以A1及A2較豐。本研究用HPLC及CE分析厚朴藥材：HPLC的P法，以流動相(a) 20 mM CH3COONH4及(b) CH3CN/H2O = 80/20(v/v)為沖提液，使用Cosmosil 5C18-MS為層析管柱，可在45分鐘內分析M及H兩酚類成分；A法則以 (a) buffer / CH3CN＝60/40（buffer為50 mM NaOAc及5 mM SDS之混合液，以10﹪AcOH調至pH=3.63）及(b) CH3OH /CH3CN /H2O = 45 /45 /10(v/v)為沖提液，可在50分鐘內同時定量酚類H、M及生物鹼A1、A2等四成分。CE則用電壓注射樣品，以負電壓進行掃集，緩衝溶液為50mM磷酸、100mM SDS與10mM diethylamine溶液，及氰甲烷( 75：25,v/v)，在此條件下放大倍數可達6.4 ×103，於45分鐘內可檢測出七個厚朴生物鹼成分吸收峰。
市售厚朴來源極為複雜，除正品的川厚朴、凹葉厚朴及和厚朴外，並有非藥用部位樣品或偽品在市場上流通。本研究從台灣及日本市場收集32批藥材，用HPLC分析M、H、A1及A2含量，用CE掃集圖譜比對其批次間成分差異，發現各厚朴樣品中的酚類含量差異不大，但M/H及A1/A2比值則有明顯區隔，以M/H比值來看：凹葉厚朴＞4.59，川厚朴＜1.41，和厚朴則介於3.12~3.91之間；至於A1/A2比值：和厚朴＞20.09是三者中最大者，凹葉厚朴＜1.98，川厚朴則介於2.16~9.81之間。利用CE掃集法可區別厚朴正品與偽品，兩者的掃集峰圖紋迥異。正品厚朴樣品有一致的七個掃集峰，而混用藥材掃集結果有二：一是圖紋為成群交雜的重疊小峰，含量較少，看不出正品吸收峰的相對應成分；一是除正品七個掃集峰外，另有其他成分吸收峰夾雜或重疊其中。推測前者為偽品，後者為非藥用部位樣品。因此，結合LC定量及CE掃集圖譜不但可用來分辨厚朴藥材基原，且可用於辨識厚朴偽品的摻雜與混淆。
市售吳茱萸藥材分吳茱萸和石虎兩品種，前本實驗室化學分析結果顯示，兩者在成分含量上沒有差異，但成熟與未成熟樣品則有明顯不同，前者有較高的生物鹼含量，後者則有較多的黃酮類成分。檢視市售樣品，發現同批藥材在成熟度、顏色、顆粒大小方面就有很大差別，本文的目的在開發一種以二或三成分辨識這些樣品的方法。首先依外觀性狀將十四批樣品分類，利用HPLC方法，以流動相(a) buffer / CH3CN＝80/20 (buffer為30 mM NaOAc與1.25﹪AcOH混合溶液，pH=3.68 )及(b) CH3OH /CH3CN /H2O /
AcOH = 45 /45 /10/0.25 (v/v)沖提分析，發現開口樣品之DE與E的總含量大於閉口樣品，而且以色黑、顆粒大、成花狀者為佳；然而，開口者的DE/E比值（0.699～2.460）卻小於閉口者（1.672～11.541）。文獻記載，具有類似腎上腺素功能的Synephrine與生長期有直接關連，分析結果顯示，不同批次間的差異極大，但同一批樣品中，閉口者總大於開口者。傳統上，吳茱萸以陳者良，本研究將吳茱萸樣品置於不同溫度、時間及環境中，發現高溫環境下藥效成分含量變動較大。室溫下的吳茱萸樣品，DE與E衰減速率緩慢且穩定，DE/E比例幾乎維持一定值；置於50℃烘箱之開口吳茱萸DE/E比值漸增，閉口者漸減，由於閉口樣品的DE快速遞減，其藥效成分比例與開口樣品漸趨一致。將以吳茱萸為君藥的吳茱萸湯製劑，存放於室溫下，發現藥效成分含量沒有太大的變動；而在高溫環境中DE與E的含量變化明顯減少，其衰變趨勢與開口吳茱萸藥材極為相似，其中Rutaecarpine與Evocarpine有先減後增的傾向，含量變化最小。

Both high-performance liquid chromatography and capillary electrophoresis are belonging to the liquid chromatographic technology and have several separation modes each. Currently they have become the most commonly used methods for analyzing the marker substances of Chinese herbal drugs. This study employed these two methods in analyzing the Chinese herb drugs Magnoliae Cortex and Evodiae Fructus. Owing to the fact that LC-UV and CE-UV have limited detecting capability, there are difficult to apply them to the analysis of animal blood composition, especially those samples with very minute amounts of constituents. This study has developed the CE-Sweeping method, by which we can successfully determine the ephedra (Ephedra Herba) alkaloids in experimental mice’s blood and the trace magnolia (Magnoliae Cortex) alkaloids in a crude drug.
Ephedra Herba has six major active constituents, namely l-ephedrine (E), d-pseudoephedrine (PE), l-norephedrine (NE), d-norpseudoephedrine (NPE), l-methylephedrine (ME) and d-methylpseudoephedrine (MPE). By means of the cation-selective exhaustive injection and sweeping (CSEI-Sweeping) method, the detectability limit can reach 20.0 ~ 99.5 ppb. This study used a buffer consisting of 25 mM phosphate solution of 50 mM SDS and acetonitrile (80 : 20, v/v) which can separate 6 constituent peaks successfully within 27 minutes with good reproducibility having an RSD.value of 1.06 ~ 2.86%. This method can be applied to analyze the contents of constituents in the mouse blood at very low concentrations.
Magnoliae Cortex is a commonly used Chinese herb drug, which possesses the effects of warming the body center, settling down chi, drying up dampness, and resolving phlegm. It contains phenols such as magnolol (M) and honokiol (H) and trace alkaloids such as magnocurarine (A1), salicifoline (A2), magnoflorine, anonaine, michelarbine, liriodenine and laurifoline,etc. Among these alkaloids, A1 and A2 exist in higher amounts. This study used HPLC and CE to analyze the drug material magnolia. With HPLC, method P used a mobile phase composed of (a) 20 mM CH3COONH4 and (b) CH3CN/H2O = 80/20 (v/v) as the eluent, and Cosmosil 5C18-MS as the chromatographic column to separate the two phenols M and H within 45 minutes; whereas method A used an eluent composed of (a) buffer/CH3CN = 60/40 (the buffer is a mixture of 50 mM NaOAc and 5 mM SDS, which is adjusted to a pH=3.63 with AcOH) and (b) CH3OH/CH3CN/H2O = 45/45/10 (v/v) to simultaneously assay the four constituents: phenols H and M and alkaloids A1 and A2 within 50 minutes.With CE, the electrokinetic mode-injected sample was sweep run under a negative voltage, using a buffer composed of 50 mM phosphoric acid, 100 mM SDS and 10 mM diethylamine: acetonitrile = 75/25 (v/v). Under this condition, the magnification factor can reach 6.4 x 103, and the absorption peaks corresponding to seven magnolia alkaloids were separated within 45 minutes.
The commercial sources of Magnolia Cortex are very complicated. In addition to the orthodox articles derived from Magnolia officinalis, M. officinalis var. biloba and M. obovata respectively, there are also articles derived from different non-medicinal used parts and counterfeits. This study collect 32 samples from the herb markets in Taiwan and Japan, and used HPLC to analyze the contents of M, H, A1 and A2 and used CE to compare the differences in trace alkaloids among different batches. As a result, it was found that the phenolic constituents did not vary very much among the various articles of Magnolia Cortex. However, the M/H and A1/A2 ratios showed marked distinctions. The M/H ratio was > 4.59 in M. offcinalis var. biloba, <1.41 in M. officinalis, and 3.12 ~ 3.91 in M. obovata. As for the A1/A2 ratio, it was >20.09 in M. obovata, the higher of all; <1.98 in M. offcinalis var. biloba; and 2.16 ~ 9.81 in M. officinalis. Using the CE-Sweeping method, we can discriminate the orthodox articles of Magnolia Cortex from the counterfeits. The sweeping patterns of both groups are drastically different. The orthodox articles showed seven sweeping peaks, whereas the counterfeits were of two groups. One group showed overlapped clusters of small peaks in lesser quantities, showing no peaks corresponding to those in the orthodox articles. The other group showed other peaks admixed with the seven sweeping peaks, showing the presence of other constituents. It is postulated that articles of the former group are counterfeits, and those of the latter group are derived from non-medicinal used parts. Hence, the combined use of LC assay and CE sweeping patterns can be applied to the identification of magnolia sources as well as to the discrimination of the orthodox articles from the counterfeits or adulterants.
Commercial articles of evodia (Evodia Fructus) are divided into two kinds: those derived from Evodia rutaecarpa and from E. rutaecarpa var. officinalis. Both kinds do not differ from each other in terms of their constituent contents. However, there is marked difference between the mature and immature samples. The former contain higher alkaloid contents, and the latter contain higher flavones contents. Examination of the commercial samples also found marked differences in maturity, color and size of fruits among samples from the same batch. This study is aimed to develop a method whereby we can identify these samples in terms of two or three constituents. First we sorted the samples by their external appearance, and then analyzed them by HPLC, using a mobile phase composed of (a) buffer/CH3CN == 80/20 (the buffer is a mixture of 30 mMNaOAc and 1.25% AcOH, pH = 3.68) and (b) CH3OH/CH3CN/H2O/AcOH = 45/45/10/0.25 (v/v). As a result, it was found that the total of DE and E contents in the open-mouth articles were larger than those in the closed-mouth articles, and that those articles with a darker color, large sizes and flowery shapes were better. However, the DE/E ratio in the open-mouth samples (0.699 ~ 2.460) was smaller than that in the closed-mouth ones (1.672 ~ 11.541). Literature records indicate some linkage between the herb’s synephrine effect and the herb’s growth age. Analysis results showed great differences among different batches, but within the same batch the closed-mouth samples always have higher content than the open-mouth ones in quantity. Traditionally, the aged article of the herb is said to be of superior quality. This study put the evodia samples at different temperatures and in different environments for different lengths of time. It was found that at high temperatures, there were greater changes in constituents. At room temperature, deterioration in DE and E was slow and stable and the DE/E ratio almost remained constant. Put in a 50℃ oven, this ratio increased gradually in the open-mouth samples, but decreased gradually in the closed-mouth samples owing to rapid reduction in DE in the closed-mouth samples whose medicinal constituents gradually approached the same as those of the open-mouth samples. By storing the herbal formula Wu-chu-yu-tang (Evodia Combination), which contains Evodiae Fructus as its imperial ingredient, at room temperature, its medicinally active constituents were found to have no noticeable changes. However at high temperatures, the DE and E contents decreased markedly in a manner similar to that of the open-mouth samples of Evodiae Fructus. Among the constituents rutaecarpine and evodarpine showed a first decrease and then increase tendency in content change, which was the minimum of all.

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