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HPLC法测定各沪产地龙和广地龙中次黄嘌呤、黄嘌呤、尿嘧啶和尿苷的含量
季倩1,高守红2,张汉明1,张磊1*
0
(1. 第二军医大学药学院药用植物学教研室, 上海 200433;
2. 第二军医大学长征医院药材科, 上海 200003
*通信作者)
摘要:
目的 测定各种沪产地龙与广地龙中次黄嘌呤、黄嘌呤、尿嘧啶、尿苷的含量,建立地龙药材质量评价的方法。方法 用0.9%生理盐水超声提取地龙, SORBAX SB-Aq色谱柱(250 mm×4.6 mm, 5 μm,Aglient Co.,Ltd),流动相:5 mmol/L 磷酸二氢钾(pH 2.9),流速1 mL/min,检测波长254 nm,柱温30℃,进样量10 μL,采用HPLC法,同时测定次黄嘌呤、黄嘌呤、尿嘧啶、尿苷4种核苷类成分的含量。结果 次黄嘌呤的线性范围为0.500 0~100.00 μg (r=0.999 9),平均回收率99.37%,RSD=1.36% (n=6);黄嘌呤的线性范围为0.500 0~100.00 μg (r=0.993 1),平均回收率 91.57%,RSD=1.40% (n=6);尿嘧啶的线性范围为0.500 0~100.00 μg (r=0.999 9),平均回收率95.31%,RSD=1.64% (n=6);尿苷的线性范围为0.500 0~100.00 μg (r=0.999 9),平均回收率100.21%,RSD=1.98% (n=6)。结论 该方法重复性、回收率好,可用于测定地龙与广地龙中次黄嘌呤等4种核苷类成分的含量。
关键词:  沪地龙  广地龙  次黄嘌呤  黄嘌呤  尿嘧啶  尿苷  高压液相色谱法
DOI:10.3724/SP.J.1008.2015.00443
投稿时间:2014-08-28修订日期:2014-11-01
基金项目:国家发展和改革委员会立项项目[发改办高技[2011]51号],上海市科委产学研医合作研究项目(13DZ1970403).
HPLC in determination of hypoxanthine, xanthine, uridine and uracil in Shanghai Pheretima and Pheretima aspergillum (E. Perrier)
JI Qian1,GAO Shou-hong2,ZHANG Han-ming1,ZHANG Lei1*
(1. Department of Pharmaceutical Botany, School of Pharmacy, Second Military Medical University, Shanghai 200433, China;
2. Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
*Corresponding author)
Abstract:
Objective To establish an effective method for the determining hypoxanthine, xanthine, uridine and uracil in earthworm in Shanghai Pheretima and Pheretima aspergillum (E. Perrier), contributing to quality control of the medicinal material. Methods Hypoxanthine, xanthine, uridine and uracil were extracted from the earthworms with 0.9% NaCl by ultrasonic and determined by HPLC. The chromatographic conditions were: SORBAX SB-Aq column (250 mm×4.6 mm, 5 μm, Aglient Co.,Ltd), 5 mmol/L KH2PO4 (pH 2.9) as the mobile phase with a flow rate of 1 mL/min,the detection wavelength was 254nm, the column temperature was set at 30℃, and the injection volume was 10 μL. Hypoxanthine, xanthine, uridine and uracil were determined by HPLC at the same time. Results The linear range was 0.5-100 μg (r=0.999 9) for hypoxanthine, with the average recovery being 99.37%, RSD=1.36% (n=6). The linear range was 0.5-100 μg (r=0.993 1) for xanthine, with the average recovery being 91.57%, RSD=1.40% (n=6). The linear range was 0.5-100 μg (r=0.999 9) for uridine, with the average recovery being 95.31%, RSD=1.64% (n=6). The linear range was 0.5-100 μg (r=0.999 9) for uracil, with the average recovery being 100.21%, RSD=1.98% (n=6). Conclusion The current method is reproducible and has satisfactory recovery, and it can be used to determine hypoxanthine, xanthine, uridine and uracil in earthworm medicinal material.
Key words:  Shanghai Pheretima  Pheretima aspergillum (E.Perrier)  hypoxanthine  xanthine  uridine  uracil  high pressure liquid chromatography