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Guizhou middle tube flowers in Codonopsis Codonopsis acetylene glycosides by HPLC_595

By Greg Armstrong,2014-10-30 19:01
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Guizhou middle tube flowers in Codonopsis Codonopsis acetylene glycosides by HPLC_595

    Guizhou middle tube flowers in Codonopsis Codonopsis acetylene glycosides by HPLC

     Abstract Objective To determine pipe flower in Guizhou in Codonopsis Codonopsis acetylene glycoside content, in order to control the quality control of Radix Codonopsis flowers and

    cultivation provide a scientific basis and so on. Methods using high performance liquid chromatography (HPLC) method analysis column according to Litt-ODS2 (250 mm × 4.6 mm, 5 μ

    m), with acetonitrile - water (20:80) as mobile phase,

    detection wavelength of 270 nm . The results determined the origin of the five tubes in flowers Codonopsis Codonopsis acetylene glycoside content, content in the range of 1.265 ~ 4.093 mg * g-1. Conclusion The method is simple, rapid, reproducible, and can be used to control spend Codonopsis the quality evaluation.

     Key Words pipe flowers Codonopsis; Codonopsis acetylene glycosides; high-performance liquid chromatography

     Determination of Lobetyolin in Radix Codonopsis Tubulosae of Guizhou

     Abstract: ObjectiveDetermination of lobetyolin in Radix Codonopsis Tubulosae of Guizhou was carried on to control the quality, introduction and cultivation, etc.MethodsHypersil ODS2 (250 mm × 4.6 mm, 5 μm) was used with acetonitrile-

    water (20:80) as the mobile phase and DAD detection was at 270 nm.ResultsRadix Codonopsis Tubulosae of 5 habitat in Guizhou were analyzed.The content of lobetyolin ranged from 1.265 ~ 4.093 mg * g-1.ConclusionThe method appeared to be simple, quick and reproducible. It can be used to control the quality of Radix Codonopsis Tubulosae.

     Key words: Radix Codonopsis Tubulosae; Lobetyolin; HPLC

     Tube Flower Codonopsis Codonopsis tubulosa Kom. For the Campanulaceae Codonopsis species, located in northwest Guizhou

    and Yunnan, Sichuan and other places. Its dried roots used as medicine, also known as the "Guizhou Codonopsis" "Syria Party," "Sweet Party" and so on, for "Guizhou Province, Chinese herbal medicines, national quality standards for medicines," 2003 Edition containing relevant species [1]. Medicine has its roots Bu Zhong Yi Qi, spleen Yifei and other functions, for the spleen and stomach weakness, shortness of breath, palpitations, eat less then pond, virtual asthma cough, heat Diabetes embolism. And some areas in Guizhou, the

    management has also been made to spend Codonopsis Codonopsis medicine use. As in the past the quality control of Radix Codonopsis there is no unified and standardized, thus affecting the effective use of Codonopsis. The 2005 edition of

    "Chinese Pharmacopoeia" a new addition to the characteristic Codonopsis acetylene glycoside constituents TLC Codonopsis [2]. Another reference [3] reported that acetylene glycosides as Codonopsis Codonopsis protect gastric mucosa, one of the active ingredient and can be used HPLC assay method. Through the literature search, no application of HPLC determination of tube flowers Codonopsis Codonopsis acetylene glycosides in relevant reports. Therefore, in order to study management in Guizhou to spend quality Codonopsis, we were determined the origin of Guizhou 5 tube flowers in Codonopsis Codonopsis acetylene glycoside content with a view to control the quality evaluation of Radix Codonopsis flowers and cultivation, etc. provide valuable reference.

     An instrument, reagents and test drug

     1.1 Instrument Agilent 1100 high-performance liquid

    chromatography; DAD detector; CSF 1B ultrasonic cleaning

    instrument (Shanghai Ultrasonic Instrument Factory); AG135 Electronic Balance (Mettler-Toledo Inc., Switzerland).

     1.2 Reagent Codonopsis acetylene glycoside (lobetyolin) reference standards (purchased in Jiangxi Heavenly Creations Herbal Technology Co., Ltd., the purity of "98%); acetonitrile for chromatography pure; methanol AR; water re-distilled

water.

     1.3 for the test to all the tested medicinal herbs by the author in person to the farms to carry out dredging by the

    Guiyang Medical College Professor He Shunzhi identified, after some back to wash, dry. Herbal Table 1.

     2 Methods and Results

     2.1 The choice of detection wavelength take appropriate Codonopsis acetylene glycoside reference substance, add

    methanol into the preparation of a certain concentration of reference substance solution into the sample, using DAD detector at 190 ~ 400 nm for detection. Codonopsis acetylene glycosides at 270 nm at a maximum absorption value, so choose

    270 nm as detection wavelength.

     Table 1 Guizhou tube flowers in Codonopsis Codonopsis acetylene glycoside content (abbreviated)

     2.2 The chromatographic conditions were column was under little ODS2 (250 mm × 4.6 mm, 5 μm); mobile phase of

    acetonitrile - water (20:80); flow rate 1 ml * min-1;

    detection wavelength 270 nm; column temperature 25 ?;

    injection volume 10 μl.

     2.3 Preparation of Solution

     2.3.1 Preparation of reference substance solution to take Codonopsis acetylene glycosides suitable reference substance, precision, said set, plus methanol produced per ml Codonopsis acetylene glycosides 0.1 mg of reference substance solution,

    that is, too.

     2.3.2 Preparation for the test product solution

     Extraction conditions to determine: methanol as the solvent, respectively bath reflux extraction, ultrasonic extraction, Soxhlet extraction, the cold soak overnight and

    other extraction methods were compared. The results showed that methanol ultrasonic extraction and reflux extraction water bath, the most complete, but the two kinds of extraction

    methods there was no significant difference, so choose ultrasonic extraction, and ultrasonic extraction time to conduct investigations. The results for the ultrasonic extraction 60 min and 120 min no significant difference, so choose ultrasonic extraction 60 min. Reposted elsewhere in the paper for free download http://

     Preparation of test items for the solution: Take the tested medicinal powder 1 g (over 3 screens), precision, said determined set Cypriot conical bottle with precision by adding methanol, 50 ml, said that given the weight, ultrasonic

    extraction 60 min, put cold, add methanol by weight loss supplement, shake, filtration. Precision continued to take the amount of filtrate 25 ml, set evaporating dish, the water bath evaporated, dissolved in methanol and quantitatively transferred to 5 ml Liang Ping, add methanol was diluted to scale, shake, using microporous membrane (0.45 μm)

    filtration , take filtrate added, that was.

     2.4 linear relationship between the inspection to take Codonopsis acetylene glycoside reference substance about 12

    mg, precision that set, and placing 10 ml Liang Ping, by using methanol dissolved and diluted to the scale, shake, draw the precise amount of solution were made 618,247.2 ,

    98.88,39.55,15.82,6.33,2.53 μg * ml-1. Respectively, the

    above reference substance solution into the sample, sample

    volume 10 μl, measured peak area. To the mass concentration of reference substance abscissa and peak area of the vertical axis, linear regression, was the regression equation Y = 9.370 07X +7.206 03, r = 0.999 99, Codonopsis acetylene glycosides

    in 0.0253 ~ 6.18 μg and the peak area between the showed good linear relationship.

     Taking a concentration of 2.5 precision experiments 247.2μg * ml-1 of the reference substance solution, a continuous injection six times, sample volume 10 μl, determined. With

    Codonopsis acetylene glycoside peak area of the indexes, RSD 0.48%, indicating good precision.

     2.6 Stability Experiment No. 1 for the test to take

    medicine powder 1 g, precision that will, according to "2.3.2" method under preparation for the test materials were solution, at room temperature, respectively 0,2,4,8,16 , 24,48 h sample measured in order to Codonopsis acetylene glycoside absorption peak area of the indexes, RSD was 0.61%, indicating that for the test solution in Codonopsis acetylene glycoside in the 48h period was basically stable.

     2.7 to reproduce the experiment to take on the 1st test

    medicinal powders for the six copies, each 1 g, precision that will, according to "2.3.2" prepared under the law, measurement, calculation content of the sample, RSD 2.45%, indicating that reproduce was good.

     2.8 The average recovery for the experiment to take on the 1st test medicinal powders six copies, each 1 g, precision that will, respectively, by adding a certain amount of Codonopsis acetylene glycoside reference substance, according to "2.3.2" prepared under the law, testing , calculate the

    recovery rate, an average recovery of 98.36%, RSD was 0.61%.

     2.9 Determination of the sample according to "2.3.2" under prepared for the test materials and reference substance solution, collected nine of the flower tube of Codonopsis

    Codonopsis herbal medicine samples for determination of acetylene glycoside, the results shown in Figure 1 and Table 1 .

     3 Discussion

     According to literature search, the application of HPLC to the tube to spend Codonopsis Codonopsis acetylene

    glycosides were determined in the first time. The experimental results show that the test tube to spend the root of Codonopsis Codonopsis acetylene glycoside content of the range of 1.265 ~ 4.093 mg * g-1, much higher than the "Chinese

    Pharmacopoeia" collection contained in the three varieties Codonopsis Codonopsis pilosula (Franch.) Nannf., Sichuan Codonopsis C.tangshen Oliv. and the Su-Hua Codonopsis

    C.pilosula var.modesta (Nannf.) LTShen in the acetylenic

    glucoside content of Codonopsis 0.035 ~ 1.314 mg * g-1 [2]. It

    was also reported Codonopsis acetylene glycosides on gastric

    mucosal damage caused by alcohol have a good protective effect is to protect the gastric mucosa Codonopsis is one of the active ingredients [2], that pipes to spend in the treatment

    of gastric ulcer diseases such as Codonopsis has more of high research value.

     Figure 1 Codonopsis acetylene glycoside (a) pipe flowers Codonopsis root (b) tube flower stems and leaves Codonopsis (c) HPLC chromatogram (omitted)

     Experimental results show that the tube flowers in Codonopsis Codonopsis acetylene glycoside content and origin related Weining produced the highest, followed by Qingzhen produced, respectively, 4.093 and 3.071 mg * g-1. In addition

    to this, Codonopsis acetylene glycoside content may change with changes in harvest time, but the exact impact remains to be further studied.

     Through two tubes the origin of leaves and stems of flowers Codonopsis Codonopsis acetylene glycoside were

    determined, the results showed the measured acetylene glycosides in leaves and stems of Codonopsis concentration range of 1.237 ~ 1.904 mg * g-1, shows the same leaf can be

    used as medicine to use, which can not only ease the shortage of resources, Codonopsis, but can also bring some economic and social benefits.

     References

     [1], Guizhou Province and Drug Administration. Guizhou Province, Chinese herbal medicine national quality standards (2003 edition) [S]. Guiyang: Guizhou Science and Technology Press, 2003:280.

     [2] National Pharmacopoeia Committee. Chinese

    Pharmacopoeia [S]. Beijing: Chemical Industry Press, 2005:199.

     [3] Qing, Mr Zhu Yuan, Wang Zheng-tao, et al. Codonopsis

    Codonopsis acetylene glycosides in HPLC analysis [J]. Chinese Pharmaceutical Journal, 2005,40 (1): 56. Reposted elsewhere in

    the paper for free download http://

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