附子不同炮制品中6个乌头碱成分含量测定方法的建立及临床用药分析

Establishment of determination method for six aconitines in different processed radix aconiti carmichaeli products and clinical medication analysis

目的:建立附子炮制品中6个乌头碱成分的含量测定方法并进行临床用药分析。方法:采用高效液相色谱法测定附子炮制品中单酯型和双酯型乌头碱含量。色谱柱为岛津Shim-pack GIST C_(18 )(250 mm×4.6 mm,5μm),流动相为乙腈-0.2%冰乙酸溶液(用三乙胺调节pH值至6.2)(梯度洗脱),流速为1.0 mL·min^(-1),检测波长为235 nm,柱温为30℃。结果:苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、乌头碱、次乌头碱的进样量线性范围分别为0.034 4~8.602 4μg(r=0.999 9)、0.003 4~0.848 9μg(r=0.999 9)、0.003 1~0.786 9μg(r=1.000 0)、0.018 9~4.726 5μg(r=1.000 0)、0.019 0~4.741 4μg(r=1.000 0)、0.017 9~4.473 0μg(r=1.000 0);平均加样回收率分别为101.5%(RSD=1.23%)、96.17%(RSD=1.58%)、98.90%(RSD=1.96%)、103.1%(RSD=1.85%)、91.16%(RSD=1.47%)、95.43%(RSD=0.31%)(n=9);精密度、重复性、耐用性试验的RSD均小于2.0%。60批样品测定结果显示,有效性:白附片、黑顺片>炮附片>淡附片;安全性:炮附片>白附片>淡附片>黑顺片;综合评价:白附片>炮附片>黑顺片>淡附片。结论:所建方法可靠、准确,可同时测定附子炮制品中6个乌头碱成分的含量,并发现4种附片中白附片有效性和安全性都较好,炮附片有效性稍弱、但安全性好,黑顺片需关注其安全性,淡附片需注意其有效性,该研究为临床用药提供了参考。

OBJECTIVE To establish the content determination method for six aconitines in processed radix aconiti carmichaeli,and to conduct clinical medication analysis.METHODS HPLC method was used to detect the contents of monoester and diester aconitines in processed radix aconiti carmichaeli.The determination was performed on a Shimadzu Shim-pack GIST C_(18 )column(250 mm×4.6 mm,5 μm) with the mobile phase consisting of acetonitrile-0.2% glacial acetic acid solution(pH was adjusted to 6.2 by triethylamine) with gradient elution at the flow rate of 1.0 mL·min~(-1),the detection wavelength of 235 nm and the column temperature at 30 ℃.RESULTS The linear ranges of benzoyl neoaconitine,benzoyl aconitine,benzoyl hypoaconitine,neoaconitine,aconitine and hypoaconitine were 0.034 4-8.602 4 μg(r=0.999 9),0.003 4-0.848 9 μg(r=0.999 9),0.003 1-0.786 9 μg(r=1.000 0),0.018 9-4.726 5 μg(r=1.000 0),0.019 0-4.741 4 μg(r=1.000 0) and 0.017 9-4.473 0 μg(r=1.000 0),respectively.The average recoveries were 101.5%(RSD=1.23%),96.17%(RSD=1.58%),98.90%(RSD=1.96%),103.1%(RSD=1.85%),91.16%(RSD=1.47%) and 95.43%(RSD=0.31%)(n=9),respectively;the RSDs of precision,reproducibility and durability tests were all lower than 2.0%.The determination results of 60 samples showed that the effectiveness order was as follows:Baifupian,Heishunpian>Paofupian>Danfupian,the safety order was Paofupian>Baifupian>Danfupian>Heishunpian,and the comprehensive evaluation order was Baifupian>Paofupian>Heishunpian>Danfupian.CONCLUSION The established method is reliable and accurate,and can simultaneously determine the contents of six aconitines in processed radix aconiti carmichaeli products.It is found that among the four types of aconite tablets,the efficacy and safety of Baifupian are good,and the efficacy of Paofupian is slightly weak,while its safety is good.Attention should be paid to the safety of Heishunpian,and attention should be paid to the effectiveness of Danfupian.The results of the study provide reference for clinical medication.