全 文 ：HPLC法测定北豆根总碱粉针剂中蝙蝠葛碱含量
王栋 ,王楠 ,刘岩 ,周媛媛
(黑龙江中医药大学 ,黑龙江 哈尔滨 150040)
摘　要:采用高效液相色谱法对北豆根总碱粉针剂中蝙蝠葛碱的含量进行测定。色谱柱为 ODS-C18
柱;流动相为磷酸三乙胺缓冲液-乙腈(76∶24),流速为 1ml min ,检测波长为 281nm。经测定标准曲线的
线性范围 0.288 ～ 1.728μg ,平均回收率 98.672%,RSD%=2.241%。结果表明 ,本方法简便 ,准确 ,快速 ,
灵敏度高 ,重现性好 。
关键词:HPLC 法;北豆根粉针剂;蝙蝠葛碱含量
中图分类号:R927.2　　　文献标识码:A　　　文章编号:1002-2392(2006)01-0008-02
收稿日期:2005-10-08
作者简介:王栋(1948-),男 ,教授 ,博士研究生导师。主要研究方向为
中药药效物质基础。
　　北豆根为防己科植物蝙蝠葛(Menispermum dauri-
cum DC.)的干燥根茎 ,为常用中药 ,具清热解毒 、祛风
止痛之功效 ,其提取物临床上常用于咽喉肿痛 ,肠炎痢
疾 、风湿痹痛。近年来 ,本课题组开发研制了北豆根总
碱粉针剂 ,通过现代药理学研究证实 ,北豆根总碱粉针
剂中主要含有的以蝙蝠葛碱为主的一类酚性生物碱 ,
对脑梗塞 、心肌缺血有较好的治疗作用 。为了建立该
粉针剂的质量标准 ,参照文献[ 1 , 2] 采用高效液相色谱法
对北豆根粉针剂中的蝙蝠葛碱进行了含量测定 ,为北
豆根粉针剂质量标准的建立提供依据。
1　仪器 、试剂和样品
1.1　仪器
日本岛津 LC-2010型高效液相色谱仪 ,操作系统
Class-vp release 6.12 SP5;日本岛津 UV-1601紫外分
光光度仪 SHIMADZU。
1.2　试剂
乙腈 ,色谱纯 ,Dikma Pure Acetonitrile 50101;DIMA
TECHNOLOGY INC(USA);磷酸 ,色三乙谱纯 ,色谱纯;
水为娃哈哈纯净水 ,过 0.45μm 水系微孔滤膜 。
1.3　对照品和样品
蝙蝠葛碱对照品:自制(纯度:99.019%)。
供试品:北豆根总碱粉针剂试品 。批号:040301 ,
040302 ,040303 ,040304 ,040305 ,040306。
2　方法和结果
2.1　色谱条件
色谱柱:ODS -C18柱(Hypersil ODS2 , 4.6mm ×
150mm ,5μm);流动相:缓冲液(将0.68ml磷酸和0.71ml
的三乙胺加纯净水至 500ml)-乙腈(76∶24);检测波
长:281nm;柱温 40℃;流速 1.0ml min;进样量 10μl。
2.2　方法学的考察
2.2.1　对照品溶液的制备
精密称取对照品 0.0144g ,置于 100ml容量瓶内加
缓冲液溶解并定容 ,浓度 0.144mg ml ,过 0.45μm水系
微孔滤膜备用。
2.2.2　供试品溶液的制备
精密称取每批样品 5mg 置于 25ml容量瓶中 ,以缓
冲液溶解并定容 ,过 0.45μm水系微孔滤膜备用。
2.2.3　线性关系
取对照品溶液分别进样 2 、4 、6 、8 、10 、12μl ,按上述
色谱条件进行测定 ,HPLC图谱如图 1 ,记录峰面积 ,以
进样量(质量)为横坐标 ,峰面积积分值为纵坐标 ,绘制
标准曲线 。结果表明蝙蝠葛碱在 0.288 ～ 1.728mg 质
量浓度范围内呈线性关系。回归方程为:Y=725474X
+2909.43　r=0.9999。
2.2.4　精密度试验
取对照品溶液 10μl ,按上述色谱条件重复进样 5
次 ,测定并记录其峰面积的积分值 ,RSD=0.411%。
2.2.5　重现性试验
·8· ACMP.Aug , 2006 , Vol.34 , No.1
图 1　蝙蝠葛碱对照品的HPLC 图
　　取同一批制剂 ,按 2.2.2方法制备 5份供试品溶
液 ,平行测定并记录其峰面积的积分值 , RSD =
0.126%。
2.2.6　稳定性试验
按2.2.2方法制备供试品溶液 ,取同一供试品溶
液每隔 2h测定 1次 ,共测定 5次 ,记录其峰面积的积
分值 ,RSD=0.303%。
2.2.7　回收率试验
取已知浓度的各批制剂的供试品溶液各 4ml ,分
别加入定量的蝙蝠葛碱对照品 ,测定结果见表 1。
表 1　加样回收率试验结果(%)(n=5)
批号 样品量(mg)
对照品加
入量(mg)
测得量
(mg)
回收率
(%)
平均回收率
(%) RSD(%)
1 0.6497 0.6480 1.2940 99.43
2 0.6448 0.6480 1.2814 98.24
3 0.6002 0.6048 1.1786 95.63 98.672 2.241
4 0.6363 0.6480 1.2734 98.32
5 0.6181 0.6048 1.2334 101.74
2.3　含量测定
将 6批北豆根总碱粉针剂样品制成溶液 ,以上述
色谱条件对其含量进行测定 ,样品的 HPLC 如图 2 ,含
量测定结果见表 2。
3　讨论
3.1　流动相的选择　参考相关文献[ 1-4] 将流动相定
为磷酸三乙胺缓冲液-甲醇 ,磷酸三乙胺缓冲液-乙
腈或磷酸三乙胺缓冲液-水。经试验对比最终确定选
择磷酸三乙胺缓冲液-乙腈(76∶24)作为流动相 ,该条
件下蝙蝠葛碱的分离度适宜。
表 2　制剂的含量测定(%)
含量 1 含量 2 平均值
040301 72.76 72.80 72.78
040302 72.53 72.45 72.49
040303 73.18 73.28 73.23
040304 72.27 72.55 72.41
040305 71.32 71.18 71.25
040306 71.86 71.82 71.84
图2　北豆根总碱粉针剂样品的 HPLC 图
3.2　检测波长的测定 　采用紫外分光光度仪在 200
～ 400nm下对对照品进行扫描 ,结果表明其在 281nm
处有最大吸收 ,故将 281nm确定为其检测波长。
3.3　结论　采用 HPLC 法测定北豆根粉针剂中蝙蝠
葛碱的含量方法简便 、准确 、快速 、可行 。由测定结果
可知 ,北豆根总碱粉针剂中蝙蝠葛碱的含量均超过
70%。该结果为北豆根总碱粉针剂的质量标准的确定
提供了可靠依据 。
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[ 2] 　GolkiewiczW , Kuczynskij.High-performanceliquidchro-matographyof-
somealkaloidsonunmodif iedsi li cagel with a queous-organics olventmix-
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[ 3] 　周桂生 ,刘乃强 ,孙信功.硅胶柱准离子交换色谱法测定马钱子中
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·9·中医药学报 2006 年第 34卷第 1 期
ABSTRACTS FROM ORIGINAL ARTICLES
Study on the preparation technique of GeQin microcapsule
MA Ying-Li1 , CUI Ming-yu1 , ZHAO Ying2
Abstract:Object:To study preparation technique of GeQin microcap-
sule.Methods:GeQin microcapsule was prepared by simple coacervation
method.Using the uniform design , the preparation technique of GeQin
microcapsule was optimized , and the quality was evaluated.Results:The
optimum preparation technique of GeQin microcapsule:the ratio of core
material to coating material was 1:2 , the temperature of encapsulation
was at 55℃ and the stirring speed was 1200r min.Conclusion:The re-
sults showed that GeQin microcapsule had appropriate diameter and dis-
tribution of microcapsule particles size as well as requirements.
Author s address:1.College of Pharmacy , Heilongjiang University of
Chinese Medicine;2.College of Pharmacy , Harbin University of Com-
merce ,Heilongjiang Harbin China, 150040
Key words:Microcapsule;Simple coacervation;Uniform design
(Original article on page 6)
Determination of Dauricine in Rhizoma Menispermi powder injec-
tion by HLPC
WANG-Dong , WANG-Nan , LIU-Yan , et al
Abstract:To establish the determination method of Dauricine in Rhizo-
ma Menispermi powder injection by HLPC by using ODS-C18 colum
(Hypersil ODS2 , 4.6mm×150mm , 5μm).The consisted of phosphate
and triethylamine-acetonierile(76:24)was used as mobile phase.The
flow rate was 1.0ml min and the wavelength of detector was 281nm.
The calibration curve was linear in the range of 0.288～ 1.728μg .The
average recovery is 98.672% and RSD=2.241%.It is seemed that
the method is simple , accurate , rapid and with good reproducibility.
Author s address:Heilongjiang University of Traditional Medicine ,
Heilongjiang ,Harbin , 150040
Key words:HLPC;RhizomaMenispermi powder injection;Dauricine de-
termination
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GUAN Qing-xia , LV Shao-wa , CUI Shui-xia et al
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nation of notoginseng saponin R1 and ginseng saponin Rg1 in Jianxin Ta-
bl by HPLC.Methods:The notoginseng saponin R1 and ginseng saponin
Rg1 in Jianxin Tabl were separated on a symmetry shield
TM RP18 5μm ,
3.9×150mm column by methanol-water(19∶81)as a mobile phase ,
flow rate at 1.0ml min ,wavelength 203nm.Results:A linear relationship
of notoginseng saponin R1 and ginseng saponin Rg1 in Jianxin Tabl were
good at 0.36 to 3.24μg and 0.7 to 6.3μg.Conclusion:The method was
found to be simple , accurate and reliable for the determination of notog-
inseng saponin R1 and ginseng saponin Rg1 in Jianxin Tabl.
Author s address:Heilongjiang University of Traditional Chinese Medi-
cine ,Harbin China , 150040
Key words:HPLC;Jianxin Tablet;Notoginseng saponin R1;Ginseng sa-
ponin Rg1 in Jianxin Tabl
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Study on pharmacodynamics of CO2 swpercritical fluid extraction
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YANG , Bo CAO Ling , WANGXi-jun
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the positive drug.The writhing test was applied , and the times of writh-
ing reactions of mice were observed and recorded within 15min after in-
jected acetic acid.The variations of body temperature of rats in 3h were
observed in rats with fever induced by 2 , 4-dinitrophenol.Auricle tu-
mefaction was applied to observe the degree of tumefaction in auricle in-
duced by dimethy lbenzene in mice.Results:(1)Each dose group of SFE
-CO2 had analgesic effect , and with significant difference compared
with control group(P <0.05 , low dose group;P <0.01 , high dose
group), but had no significant difference compared with positive control
group(P>0.05), only a little better than positive control group.(2)
Each dose group of SFE-CO2 had the significant antipyretic effect on
the rats with the high body temperature induced by 2 , 4-dinitrophenol
within 50-180min.Compared with positive control group , the antipyret-
ic effect of SFE-CO2 is waker , but afer 50 min and 150min , the effect
of SFE-CO2 high、middle dose group and low dose group had no re-
markable difference compared with positive control group respectively
(P>0.05).(3)The SFE-CO2 high and middle dose group had anti
-inflammatior activity induced by dimethylbenzene(P<0.05), but no
remarkable difference was observed compared with positive control group
(P>0.05).Conclusion:All of above results indicated that SFE-CO2
has the effect of analgesic , antipyretic and anti-inflammatory.The SFE
-CO2 has the similar effect of analgesic and anti-inflammatory as as-
pirin , but the effect is slower in antipyretic, also , there is a relationship
between dose and effect.
Author s address:Heilongjiang University of Traditional Chinese Medi-
cine ,Harbin China , 150040
Key words:Radix Saposhinikovise;Supercitical Fluid Extraction;Phar-
macodynamics
(Original article on page 14)
Clinical study on the effect of Shenshuai Capsule on renal function
and renal fibrosis of patients in chronic renal failure
YU-mei , CHI Ji-ming , ZHANG Pei-qing , et al
Abstract:To observe the effect of Shenshuai Capsule on renal function
and renal fibrosis of patients with chronic renal failure(CRF).Meth-
ods:Effect of Shenshuai Capsule on Serum Creatinine(Scr), Blood Urea
Nitrogen(BUN), Creatinine Clearance Rate(Ccr), Fibronectin(FN),
Precollegen Ⅲ(PC -Ⅲ), CollagenⅣ(Col Ⅳ)and Hyaluronic Acid
(HA)was observed in CRF patients.Results:Compared with the control
values , Shenshuai Capsule significently decreased the level of Scr、BUN、
HA、PC-Ⅲ 、ColⅣ and elevate Ccr considerably(P<0.05).Conclu-
sion:Shenshuai Capsule could improve renal fibrosis and delay of renal
process failure.
Author s address:Heilongjiang Hospital of Traditional Chinese Medi-
cine ,Heilongjiang Harbin , 150036
Key words:Shenshuai Capsule;Chronic Renal Failure;Renal Fibrosis
(Original article on page 47)