目的建立附子中单酯及双酯型生物碱成分的高效液相色谱定量方法。方法附子干燥粉末用氨水浸润后，以异丙醇-醋酸乙酯(1∶1)超声提取30min，提取液蒸干后溶于0。01%盐酸甲醇中，经微孔滤膜滤过后，20μL进行HPLC分析。HPLC分析采用Zorbax Extend-C18柱(250mm×4。6mm，5μm)，以乙腈-40mmol/L醋酸铵缓冲液(氨水调pH10。0)为流动相进行梯度洗脱，体积流量1。0mL/min，检测波长230nm。结果6种被测生物碱成分，包括乌头碱、次乌头碱、新乌头碱、苯甲酰乌头原碱、苯甲酰次乌头原碱和苯甲酰新乌头原碱分离良好，各成分质量浓度与峰面积在测定范围内均呈良好的线性关系(r>0。999)，重现性好，6种成分的RSD(n=5)均小于1。50%;平均加样回收率(n=5)为98。20%~100。47%;最低检测限分别为0。26、0。36、0。92、0。30、0。52、0。82ng。结论本研究所建立的HPLC方法稳定可靠、简便易行，可用于附子类药材中单酯型和双酯型生物碱的同时定量分析，为附子药材及其炮制品的质量评价和质量控制奠定了基础。

Objective To establish an HPLC method for simultaneous determination of monoester-and diester aconitium alkaloids,including aconitine,hypaconitine,mesaconitine,benzoylaconine,benzoylhypaconine,and benzoylmesaconin in Chinese materia medica,Radix Aconitii Lateralis Praeparata.Methods The powdered Radix Aconitii Lateralis Praepasata material was alkalized by adding aqueous ammonia,and then extracted with a mixture solvent of isopropanol-ethyl acetate(1∶1)by ultrasonic extraction for 30 min.The extract solution was concentrated to dryness and reconstituted in methanol containing 0.01% hydrochloride acid.After filtration,the sample solution was analyzed by HPLC.The assay was performed on a Zorbax Extend-C18 column(250 mm×4.6 mm,5 μm),eluted with a mobile phase consisted of acetonitrile and 40 mmol/L ammonium acetate(pH value 10.0,adjusted with aqueous ammonia)and gradient elution at a flow rate of 1 mL/min.The eluent was monitored by a UV detector at 250 nm.Results Excellent chromatographic separation was achieved for six studied alkaloids with good linearity(r0.999)over the studied concentration ranges.RSD(n=5)were less than 1.50% and extraction recovery(n=5)were ranged from 98.20% to 100.47% for the six studied alkaloids.The limits of detection were 0.26,0.36,0.92,0.30,0.52,and 0.82 ng,respectively.Conclusion The HPLC method developed for simultaneous determination of monoester-and diester-aconitum alkaloids is simple and valid.It can be used for quality evaluation and quality control of Radix Aconitii Lateralis Praeparata and its processing products.

国家科技支撑计划项目（2006BAI09B05-2）