白及藥材的HPLC指紋圖譜建立及聚類分析
發(fā)布時間:2019-08-30 來源: 幽默笑話 點擊:
摘 要 目的:建立白及藥材的高效液相色譜(HPLC)指紋圖譜,并對樣品進行聚類。方法:采用HPLC法,色譜柱為Diamonsil C18,流動相為0.1%磷酸溶液-乙腈(梯度洗脫),流速為1.0 mL/min,檢測波長為270 nm,柱溫為30 ℃,進樣量為10 μL。以1,4-二[4-(葡萄糖氧)芐基]-2-異丁基蘋果酸酯為參照,繪制10批藥材樣品的HPLC圖譜,采用《中藥色譜指紋圖譜相似度評價系統(tǒng)》(2004 A版)進行相似度評價,確定共有峰,并采用SPSS 23.0統(tǒng)計軟件對其進行聚類分析。結(jié)果:10批藥材樣品的HPLC圖譜有15個共有峰,其中9批相似度均大于0.90;經(jīng)驗證,10批藥材樣品HPLC圖譜與對照指紋圖譜具有較好的一致性。10批藥材樣品可聚為3類,S1、S2、S3、S4、S6、S8、S9、S10聚為第1類,S7聚為第2類,S5為第3類。結(jié)論:所建HPLC指紋圖譜和聚類分析結(jié)果可為白及藥材的質(zhì)量控制提供參考。
關(guān)鍵詞 白及;高效液相色譜法;1,4-二[4-(葡萄糖氧)芐基]-2-異丁基蘋果酸酯;指紋圖譜;聚類分析
中圖分類號 R927.2;R284.1 文獻標志碼 A 文章編號 1001-0408(2018)22-3050-04
DOI 10.6039/j.issn.1001-0408.2018.22.06
ABSTRACT OBJECTIVE: To establish HPLC fingerprint of Bletilla striata, and conduct cluster analysis. METHODS: HPLC method was adopted. The determination was performed on Diamonsil C18 column with mobile phase consisted of 0.1% phosphoric acid solution-acetonitrile (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 270 nm, and column temperature was 30 ℃. The sample size was 10 μL. Using 1, 4-two [4-(glucose oxy) benzyl]-2-isobutyl malate as reference, HPLC chromatograms of 10 batches of medicinal samples were determined. Similarity evaluation was conducted by using Similarity Evaluation System for Chromatographic Fingerprint of TCM (2004 A edition). Common peaks were determined, and cluster analysis was also conducted for it by using SPSS 23.0 statistic software. RESULTS: There were 15 common peaks in 10 batches of samples, and the similarities of 9 batches were more than 0.90. After validation, HPLC chromatograms of 10 batches of medicinal material were all in agreement with control fingerprint. The 10 batches of samples could be clustered into 3 categories. S1, S2, S3, S4, S6, S8, S9, S10 were clustered into category 1; S7 was clustered into category 2; S5 was clustered into category 3. CONCLUSIONS: Established fingerprint and the cluster analysis results can provide reference for the quality control of B. striata.
KEYWORDS Bletilla striata; HPLC; 1, 4-two [4-(glucose oxy) benzyl]-2-isobutyl malate; Fingerprint; Cluster analysis
白及為蘭科植物白及[Bletilla striata (Thunb.) Reiehb. f.]的干燥塊莖,其味苦、甘、澀,微寒;歸肺、肝、胃經(jīng),具有收斂止血、消腫生肌之功效,臨床多用于咯血、吐血、外傷出血、瘡瘍腫毒、皮膚皸裂等癥的治療[1],F(xiàn)代藥理學(xué)研究表明,白及具有止血[2]、抗菌[3]、抗腫瘤[4]、抗病毒[5]等多種藥理作用。白及中含有多糖類、菲類、聯(lián)菲類、聯(lián)菲醚類、聯(lián)芐類、甾體類和三萜類等化學(xué)成分[6],其中2-異丁基蘋果酸葡萄糖氧基酯類的含量較高[7]。
2015年版《中國藥典》(一部)關(guān)于白及藥材的質(zhì)量控制標準中僅有理化鑒別、薄層色譜鑒別及水分、灰分檢查等項目[1]。目前,對白及的研究主要集中在化學(xué)成分[6]、藥理作用[2-5]、含量測定[7-8]等方面。但據(jù)筆者調(diào)研所知,目前白及的野生資源十分匱乏,藥材主要源自栽培,而白及藥材質(zhì)量受生態(tài)環(huán)境的影響很大,并且市場上存在大量偽品(如黃花白及、小白及、華白及、竹葉蘭、獨蒜蘭和舌苞蘭等),使白及藥材的有效性及安全性無法得到保障[9]。僅靠一兩個化學(xué)成分的含量測定,并不能有效控制白及藥材整體質(zhì)量,而且該藥材中許多成分的對照品在國家藥品標準物質(zhì)目錄中均查詢不到,這給同時檢測其中多指標成分的含量帶來了一定難度。指紋圖譜是醫(yī)藥學(xué)界公認的控制中藥材質(zhì)量的有效方法,具有整體性和模糊性的特點。為此,筆者收集不同產(chǎn)地來源的白及藥材,建立其高效液相色譜(HPLC)指紋圖譜,并對得到的數(shù)據(jù)進行處理,結(jié)合聚類分析方法進行質(zhì)量評價,為其質(zhì)量控制提供科學(xué)依據(jù)。
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