使用高效能液相層析法分析三種植物中多酚類化合物及總酚量測定

多酚類化合物因具有抗氧化能力，是近幾年非常熱門的研究。文獻指出多酚類化合物可抗病毒或抗菌，降低人類低密度脂蛋白氧化進而預防心臟血管疾病，也被應用於癌症的治療。因此藉由分析天然物中的多酚成份，可以幫助了解及推論天然物的保健功效。本次研究使用高效液相層析儀建立13種多酚標準品的檢量線，分析萃取自三種植物的四種樣本 (DCE、AAE、SME、SMW) 中的多酚成份。高效液相層析儀搭配紫外光全波長檢測器及C-18管柱，最佳分離條件測試結果如下，流動相為0.1%甲酸搭配氰化甲烷含有0.1%甲酸，流速設定在每分鐘1毫升，偵測波長為285nm，溫度為室溫。準確度以添加回收方式進行，測得回收率為93 ~ 126 %。將樣本的層析圖中成份的積分面積帶入檢量線的線性方程式中，求得植物萃取物樣本中成份的濃度，結果呈現出DCE內含芸香苷0.94 mg/g、棉纖維素0.11 mg/g ，AAE內含阿魏酸0.13 mg/g、棉纖維素0.30 mg/g ，SME內含芸香苷0.03 mg/g ，SMW內含綠原酸0.36 mg/g。利用Folin–Ciocalteu比色法測得四種樣本的總多酚含量分別為DCE：57.74 ± 0.21 mg/g，AAE：356.47 ± 22.88 mg/g ，SME：549.29 ± 12.14 mg/g，SMW：276.79 ± 2.54 mg/g。實驗結果顯示萃取物雖然富含多酚，然而與標準品比對，只有1至2個成份吻合，未來可使用氣相層析質譜儀針對萃取物樣本中的未知尖峰進行分析，以便取得樣本萃取物更多資訊。

Polyphenolic compounds are popular studies in the present day because of their antioxidative activity. Researches have shown that polyphenolic compounds have antiseptic, anti-virus and anticancer activities, and are able to inhibit LDL oxidation and prevent cardiovascular disease. Therefore, by analyzing the polyphenols in natural products, it can help to understand and infer their health effects. In this study, 13 polyphenols were used as standards for the establishment of the calibration lines to analyze the polyphenol components of four samples extracted form three plants (DCE, AAE, SME, SMW) . High performance liquid chromatography (HPLC) equipped with a full spectrum UV detector and a C-18 column were used. The optimal condition for analysis was as follows: 0.1% formic acid and acetonitrile with 0.1 % formic acid as mobile phase, 1 mL/min flow rate, 285nm detection wavelength and room temperature. The accuracy of analysis was assessed by adding external standard to the samples and calculating the recovery rate which has to be within 93 ~ 126 % to be a good reliability. The results showed that DCE contained rutin (0.94 mg/g) and gossypin (0.11 mg/g), AAE contained ferulic acid (0.13 mg/g) and gossypin (0.30 mg/g), SME contained rutin (0.03 mg/g), and SMW contained chlorogenic acid (0.36 mg/g). The total polyphenol contents of the four samples were determined by Folin-Ciocalteu colorimetric method and the results were DCE 57.74 ± 0.21 mg/g, AAE 356.47 ± 22.88 mg/g, SME 549.29±12.14 mg/g, SMW 276.79±12.54 mg/g. The results showed that the samples are rich in polyphenols, but only 1 to 2 of the ingredients are aligned with the standards. In the future gas chromatography-mass spectrometry (GC-MS) can be used to identify the unknown peaks of the samples.