不同產(chǎn)地梅花鹿鹿茸藥材中5種核苷類成分的含量測定及聚類分析

　　摘 要 目的：建立同時測定梅花鹿鹿茸藥材中尿嘧啶、次黃嘌呤、尿苷、肌苷、鳥苷 5 種核苷類成分含量的方法，并比較不同產(chǎn)地梅花鹿鹿茸藥材中上述核苷類成分含量的差異。方法：采用高效液相色譜法。色譜柱為Agilent Eclipse Plus C18，流動相為甲醇-0.07%冰醋酸溶液（4 ∶ 96，V/V），流速為1.0 mL/min， 檢測波長為254 nm，柱溫為28 ℃，進(jìn)樣量為5 μL。采用SPSS 19.0軟件對30批不同產(chǎn)地梅花鹿鹿茸藥材（S1～S30）進(jìn)行聚類分析。結(jié)果：尿嘧啶、次黃嘌呤、尿苷、肌苷、鳥苷的檢測質(zhì)量濃度線性范圍均為0.001～0.01 mg/mL（r均為0.999 9）；定量限分別為1.489 1、1.927 9、4.880 9、7.884 6、8.092 1 ng，檢測限分別為0.446 7、0.578 4、1.464 3、2.365 4、2.427 7 ng；精密度、穩(wěn)定性、重復(fù)性試驗(yàn)的RSD均小于2%；加樣回收率分別為99.69%～103.65%（RSD=1.40%，n=9）、97.77%～103.26%（RSD=1.67%，n=9）、97.82%～101.81%（RSD=1.12%，n=9）、99.30%～104.82%（RSD=1.72%，n=9）、98.13%～100.20%（RSD=0.64%，n=9）。30批藥材樣品中尿嘧啶、次黃嘌呤、尿苷、肌苷、鳥苷的含量存在差異。上述藥材樣品可聚為3大類，即S16～S18、S22～S30聚為一類，S8～S15、S19～S21聚為一類，S1～S7聚為一類。結(jié)論：該方法精密度、穩(wěn)定性、重復(fù)性均較好，可用于同時測定梅花鹿鹿茸藥材中尿嘧啶、次黃嘌呤、尿苷、肌苷、鳥苷的含量；梅花鹿鹿茸藥材的質(zhì)量受生態(tài)環(huán)境、養(yǎng)殖技術(shù)等因素的影響，存在較大差異。
　　關(guān)鍵詞 梅花鹿鹿茸；尿嘧啶；次黃嘌呤；尿苷；肌苷；鳥苷；高效液相色譜法；含量測定；聚類分析
　　中圖分類號 R284.1 文獻(xiàn)標(biāo)志碼 A 文章編號 1001-0408（2018）14-1945-05
　　DOI 10.6039/j.issn.1001-0408.2018.14.17
　　ABSTRACT OBJECTIVE： To set up a method for simultaneous determination of 5 nucleosides as uracil， hypoxanthine， uridine， creatinine and guanosine in Cervus nippon， to compare the differences of above nucleosides content in C. nippon among different producing areas. METHODS： HPLC method was adopted. The determination was performed on Aglient Eclipse Plus C18 column with methanol-0.07% glacial acetic acid （4 ∶ 96，V/V） as mobile phase at the flow rate of 1.0 mL/min. The detection wavelength was 254 nm. The column temperature was set at 28 ℃， and sample size was 5 ?L. Cluster analysis was conducted for 30 batches of C. nippon from different producing areas by using SPSS 19.0 software. RESULTS： The linear ranges of uracil， hypoxanthine， uridine， creatinine and guanosine were 0.001-0.01 mg/mL （all r≥0.999 9）. The limits of quantitation were 1.489 1， 1.927 9， 4.880 9， 7.884 6， 8.092 1 ng， and the limits of detection were 0.446 7， 0.578 4， 1.464 3， 2.365 4， 2.427 7 ng， respectively. RSDs of precision， stability， reproducibility tests were lower than 2%. The recoveries were 99.69%-103.65%（RSD=1.40%，n=9），97.77%-103.25%（RSD=1.67%，n=9）， 97.82%-101.81%（RSD=1.12%， n=9）， 99.30%-104.82%（RSD=1.72%，n=9） and 98.13%-100.20%（RSD=0.64%，n=9）. The contents of uracil， hypoxanthine， uridine， creatinine and guanosine were significantly different in 30 batches of C. nippon， which were clustered into 3 categories. S16-S18 and S22-S30 were clustered into one category； S8-S15 and S19-S21 were clustered into one category； S1-S7 were clustered into one category. CONCLUSIONS： The method has good precision， stability and reproducibility. It is suitable for simultaneous determination of uracil， hypoxanthine， uridine， creatinine and guanosine in C. nippon. The quality of C. nippon has great difference due to ecological environment and technical factors of farming.

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