亚洲免费av电影一区二区三区,日韩爱爱视频,51精品视频一区二区三区,91视频爱爱,日韩欧美在线播放视频,中文字幕少妇AV,亚洲电影中文字幕,久久久久亚洲av成人网址,久久综合视频网站,国产在线不卡免费播放

        ?

        Establishment of a Method for Determination of Anemoside B4 Content in Pulsatilla Water Extract

        2015-02-06 08:44:20JianfangWANG
        Agricultural Science & Technology 2015年8期
        關(guān)鍵詞:水提液白頭翁藥典

        Jianfang WANG

        Beijing Key Laboratory of TCVM,Beijing University of Agriculture,Beijing 102206,China

        Establishment of a Method for Determination of Anemoside B4 Content in Pulsatilla Water Extract

        Jianfang WANG*

        Beijing Key Laboratory of TCVM,Beijing University of Agriculture,Beijing 102206,China

        [Objective]This study aimed to establish a new method for determination of anemoside B4 content in pulsatilla water extract.[Method]Using acetonitrile-water (28:72)as the mobile phase,the high performance liquid chromatography,equipped with UV detector,was used to determine the anemoside B4 content in pulsatilla water extract. [Result]In the concentration range of 300-800 μg/ml,anemoside B4 content showed a good linear relationship with peak area.The average recovery of anemoside B4 was 98.12%(n=6;RSD=1.37%).[Conclusion]The established method meets the requirements by methodology,and it can be used to determine the anemoside B4 content in pulsatilla water extract.

        Pulsatilla;Anemoside B4;High performanceliquid chromatography; Method establishment

        P ulsatillareferstothedried roots ofPulsatilla chinensis (Bge.)Regel.It is dug out in spring and autumn.After removing the sediment on the surface,the pulsatilla is dried.Pulsatilla has functions of clearing heat, detoxifying, cooling blood and curing dysentery,so it is often used to cure heat toxin,bloody dysentery,pruitus vulvae and morbid leucorrhea[1-2].In this study,the anemoside B4 content in pulsatilla water extract was determined by high performance liquid chromatography,which is a determination method with strong specificity and reproducibility.

        Materials and Methods

        Materials

        The used equipment and reagents included high performance liquid chromatography(Waters Technology(Shanghai)Co.,Ltd.),ultrapure water(Shanghai Lixin Instrument Co., Ltd.),microporous membrane(Tianjin Keyilong Experimental Equipment Co., Ltd.),anemoside B4 (Batch Number, E-0675;Shanghai Tongtian Biotechnology Co.,Ltd.),pulsatilla(Beijing Tongrentang Co.,Ltd.;Origin,Hebei), pulsatilla water extract(prepared by the Beijing Key Laboratory of TCVM), acetonitrile and methanol(of chromatographic grade;Tianjin Guangfu Institute of Fine Chemicals).

        Methods

        Chromatography conditionsThe chromatography conditions were as follows:column,Bonna-Agela (5 μm, 150 mm×4.6 mm);mobile phase, acetonitrile-water (28∶72);flow rate, 1.0 ml/min;detection wavelength,205 nm;column temperature,25℃[3].

        Preparation of standard solutions

        A certain amount(25 mg)of anemoside B4 standard was weighed accurately and dissolved in methanol to 25 ml.The standard solution of 1 mg/ml was prepared.Certain amounts(1.5, 2.0,2.5,3.0,3.5 and 4.0 ml)of previously prepared standard solution(1 mg/ml)were diluted with methanol to 5 ml,respectively.Thus,the standard solutions of 300,400,500,600,700 and 800 μg/ml were prepared.

        Preparation of pulsatilla solutionsA certain amount (4.0 g)ofdried anemoside B4 sample was dissolvedin 100 ml of distilled water,and then heated to reflux for 60 min.The solution was cooled and filtered,and finally,90 ml of sample solution was obtained.

        Preparation of negative control solutionA certain volume(2 ml)of ultrapure water was filtered and treated as the negative control solution.

        Results and Analysis

        Methodological analysis

        System suitabilityCertain amounts (10 μl for each)of negative control solution,anemoside B4 standard solution and anemoside B4 sample solution were automatically injected into the high performance liquid chromatography.As shown in Fig.1,the peak of anemoside B4 appeared around the retention time of 8.5 min,and it was also shown on the chromatogram of anemoside B4 sample solution,but was not shown on the chromatogram of negative control solution.Under above chromatography conditions,the anemoside B4 could be separated with other components in the sample solution with a resolution of 5.87.According to the peak of anemoside B4,the theoretical plate number was calculated as 4 150.

        LinearrelationshipCertain amounts(10 μl for each)of anemoside B4 standard solutions with concentrations of 300,400,500,600,700 and 800 μg/ml were loaded automatically into the high performance liquid chromatography.After the processing,the peak area of anemoside B4 for each concentration was calculated.Treating anemoside B4 concentration(μg/ml) as the abscissa and peak area of anemoside B4 as the ordinate,the regression equation was deduced:

        y=2 426.1x+3 345.9,R2=0.999 7 (r=0.999 8).

        The results showed that within the concentration range of 300-800 μg/ml, the peak area of anemoside B4 had a good linear relationship with anemoside B4 concentration(Fig.2).

        PrecisionThe anemoside B4 concentration in the anemoside B4 standard solution was determined 6 times under the same chromatography conditions.The relative standard deviation (RSD)ofobtainedpeakareasof anemoside B4 was 0.21%,indicating the HPLC instrument has a good precision.

        ReproducibilityThe anemoside B4 concentration in the same-batch pulsatilla water extract was determined 6 times under the same chromatography conditions.The results showed that the RSD of obtained peak areas of anemoside B4 was 0.93%,indicating that the established method has a good reproducibility.

        Solution stabilityThe anemoside B4 concentration in the same-batch pulsatilla water extract was determined 0,1,4,8,12 and 24 h after the preparation respectively. The results showed that within 24 h,the RSD of peak areas of anemoside B4 was 1.03%,indicating that the prepared sample solution was stable within 24 h.SamplingrecoveryA certain amount(9 ml)of pulsatilla water extract,in which the anemoside B4 concentration was known,was mixed with a certain amount(1 ml)of anemoside B4 standard solution.The anemoside B4 concentration in the new solution was determined 6 times.The results showed that the average recovery of the 6 samples was 98.12%,and RSD was 1.37%,indicating that the established method has a good sampling recovery.

        Determination of samples

        The anemoside B4 concentration in the prepared sample solution was determined under the samechromatography conditions.The anemo-side B4 content in the sample solution was calculated by one point external standard method.The results showed that the anemoside B4 concentration in the sample solution was 579.3 μg/ml,and in the dried anemoside B4 sample was 13.03 mg/g.

        Conclusions and Discussion

        In the experiment,it was found that the column temperature showed great effect on peak flowing out of anemoside B4.At the temperature of 25℃,the retention time of anemoside B4 peak was 8.544 min,while at the temperature of 30℃,the retention time was changed to 19.301 min.In conclusion,the established method meets the requirements by methodology,and it can be used to determine the anemoside B4 content in pulsatilla water extract.

        [1]State Pharmacopoeia Commission of China(國家藥典委員會).Pharmacopoeia of the People’s Republic of China(中華人民共和國藥典(一部))[S]. Beijing:China Medical Science Press (北京:中國醫(yī)藥科技出版社),2010.

        [2]WU FY(吳鳳英),YOU BG(游本剛), TANG LH(唐麗華),et al.Determination of pulchinenoside B4 from Pulsatillae radix(白頭翁藥材中白頭翁皂苷B4的測定 )[J].Chinese TraditionalPatent Medicine(中成藥),2011,33(6):1021-1024.

        [3]QU LM(曲龍妹),ZHAO CJ(趙春杰),LI D (李丹),et al.Determination of pulchinenoside B4 in Pulsatillae decoction by RP-HPLC(RP-HPLC法測定白頭翁湯中白頭翁皂苷 B4的含量)[J].Journal of Shenyang PharmaceuticalUniversity (沈陽藥科大學學報),2009,26(1):45-47.

        Responsible editor:Tingting XU

        Responsible proofreader:Xiaoyan WU

        白頭翁水提液中白頭翁皂苷B4含量檢測方法的建立

        王建舫*
        (北京農(nóng)學院獸醫(yī)學(中醫(yī)藥)北京市重點實驗室,北京102206)

        [目的]建立白頭翁水提液中白頭翁皂苷B4含量檢測的方法。[方法]以乙腈-水(28∶72)為流動相,采用高壓液相色譜儀和紫外檢測器進行測定。[結(jié)果]在300~800 μg/ml范圍內(nèi),白頭翁皂苷B4濃度和峰面積的線性關(guān)系良好,白頭翁皂苷B4的平均回收率為98.12%(n=6),RSD為1.37%。[結(jié)論]該方法符合方法學考察的規(guī)定,可作為白頭翁水提液中白頭翁皂苷B4含量測定的方法。

        白頭翁;白頭翁皂苷B4;高效液相色譜;方法建立

        北京市教育委員會面上項目(KM201410020007)。

        王建舫(1969-),女,河北肅寧人,實驗師,從事中獸藥防治動物疾病的研究,E-mail:wjfhlx@126.com。*通訊作者。

        2015-06-05

        修回日期 2015-07-13

        Supported by GeneralProgram ofBeijing MunicipalEducation Commission (KM201410020007).

        *Corresponding author.E-mail:wjfhlx@126.com

        Received:June 5,2015 Accepted:July 13,2015

        猜你喜歡
        水提液白頭翁藥典
        白頭翁
        還有多少“穿山甲”需從藥典除名
        續(xù)斷水提液誘導(dǎo)HeLa細胞的凋亡
        中成藥(2017年12期)2018-01-19 02:06:52
        桂枝等18種中藥材水提液對5-脂肪氧化酶(5-LOX)活性的抑制作用
        2015年版《中國獸藥典》編制完成
        人參水提液通過免疫調(diào)節(jié)TAMs影響A549增殖
        中成藥(2016年8期)2016-05-17 06:08:15
        殼聚糖絮凝處理蒲地藍三味水提液效果及機理
        想改名的白頭翁
        科學啟蒙(2014年4期)2015-01-19 06:02:59
        白頭翁研究進展
        中成藥(2014年11期)2014-02-28 22:30:03
        國家藥典委員會對《中國藥典》2015年版三部增修訂內(nèi)容征求意見
        精品露脸国产偷人在视频| 色偷偷av一区二区三区人妖| 精品女人一区二区三区| 91久久精品色伊人6882| 久久精品黄aa片一区二区三区| 少妇高潮喷水久久久影院| 欧美丝袜秘书在线一区| 人妻在线有码中文字幕| 国产精品白浆在线观看免费| 一本久久a久久精品亚洲| av少妇偷窃癖在线观看| 国产一区二区三区在线影院| 伦伦影院午夜理论片| 日本丰满熟妇bbxbbxhd| 亚洲色欲大片AAA无码| 看中文字幕一区二区三区| 好吊妞视频这里有精品| 欧美人与动牲猛交xxxxbbbb| 国产亚洲欧美精品一区| 亚洲av熟女天堂久久天堂| 国产 一二三四五六| 四川老熟妇乱子xx性bbw| 婷婷第四色| 麻豆国产精品伦理视频| 精品免费国产一区二区三区四区| 性动态图av无码专区| 国产一精品一aⅴ一免费| 免费人成黄页网站在线一区二区| 特黄熟妇丰满人妻无码| 人与嘼av免费| 精品国产一区二区三区久久狼| 亚洲中文字幕在线一区| 久久人妻内射无码一区三区| 日韩国产成人精品视频| 久久精品伊人久久精品伊人| 国产免码va在线观看免费| 婷婷综合缴情亚洲| 亚洲黄片高清在线观看| 日本免费在线不卡一区二区| 国产精品夜间视频香蕉| 国产一区二区三区爆白浆|