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High Performance Liquid Chromatography Wulongdan the content of ginsenoside Rg1_304

By Ruby Sanchez,2014-10-30 19:42
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High Performance Liquid Chromatography Wulongdan the content of ginsenoside Rg1_304

    High Performance Liquid Chromatography Wulongdan the content of ginsenoside Rg1

     Abstract Objective To establish the determination of ginsenoside Rg1 Wulongdan concentration method. Method by high performance liquid chromatography the content of ginsenoside

    Rg1. Chromatographic conditions: mobile phase of acetonitrile: 0.2% phosphoric acid (83:17), detection wavelength of 203 nm, flow rate 1.0 ml / min. The results of ginsenoside Rg1 in the 0.404 ~ 4.040 μg was within the scope of a good linear

    relationship, r = 0.999 6. The average recovery rate of 96.34%, RSD = 2.37%. Conclusion This method is simple, accurate, reproducible, and can be used for the determination of ginsenoside Rg1 Wulongdan content.

     Key words ginsenoside Rg1 Wulongdan high-performance liquid

    chromatography

     Determination of Ginsenoside Rg1 in Wulongdan by HPLC

     Abstract: Objective To estabish a method for

    determination of ginsenoside Rg1 in Wulongdan.

    MethodsGinsenoside Rg1 was determined by HPLC.The mobile phase

    was acetonitrile-0.2 moL / L phosphoric acid (83:17) at a flow of 1.0 ml / min, and detection wavelength was 203nm.

    ResultsThe determination method for ginsenoside Rg1 had a good linear relationship in the range of 0.404 ~ 4.04 μg, r =

    0.999 6. The average recovery was 96.34%, RSD = 2.37%. ConclusionThis method is simple, accurate and reliable, which

    can be used for determination of the contents of ginsenoside Rg1 in Wulongdan.

     Key words: Wulongdan; Ginsenoside Rg1; HPLC

     Oolong Dan Zang, Southern Medical University, Professor Kun-tang clinical experience side, from Astragalus membranaceus, Dried ginseng, Shouwu, Kudzu, Chinese angelica, Chuanxiong and many other Chinese herbal medicines, has a Yiqihuoxue, kidney fill in the marrow, Huayu Tong Luo, etc. efficacy, clinical for the treatment of ischemic cardiovascular and cerebrovascular diseases, efficacy sure

    [1,2]. The research group early in its quality control has done a lot of work [3]. Taking into account the complexity of this product group side, Sun-dried ginseng is the main drug,

    in order to better control the inherent quality of the drug to

    ensure the safe and effective drugs, this experiment further use of high-performance liquid chromatography method for the preparation of major components of ginseng -Rg1 for

    determination.

     1 Materials and equipment

     Wulongdan liquid extract and the negative reference substance. Wulongdan Prescription: Radix Astragali 30 g, Sun-

    dried ginseng 10 g, Atractylodes macrocephala 10 g, Chinese angelica 10 g, Chuanxiong 10 g, Salvia 20 g, leech 3 g, earthworm 10 g, Gegen 30 g, the system Shouwu 20 g , Ligustrum

    lucidum 20 g, Shichangpu 10 g, Scorpio 6 g, Jiangcan 10 g (from the Southern Medical University School of Medicine in pharmacies). Prescriptions for more than a total of frying, condensed into a liquid extract (1ml extract ? 2.5 g raw

    herbs). Self-made negative reference substance. Ginsenoside Rg1 (Lot No. :0703-200015), the reagents used for

    chromatography of pure acetonitrile (USA Fisher Company); methanol chromatography pure (Tianjin Concord Technology Co., Ltd.), other reagents were analytically pure, water for pure water. Shimadzu Corporation Shimadzu high-performance liquid

    chromatography, ALS autosampler, DAD UV detector, Millennium 32 chromatography workstation.

     2 Methods and Results

     2.1 ginsenoside Rg1 Precision Preparation of reference substance solution that will ginsenoside Rg1 5.05 mg, dissolved in methanol to 25 ml Liangping constant volume, that is too.

     2.2 Preparation of sample solution precision drawing

Wulongdan 14 g, set 50 ml flask with plug in, precision by

    adding 70% ethanol to the volume, weighing, immediately ultrasound 30 min, put it aside for 10 min, make up the original weight, filtration , precision drawing continued filtrate 25 ml, decompression and recovery of ethanol to the

    dry residue dissolved in 10 ml of water, aqueous solution 50 ml separatory funnel set, the aqueous solution is too large macroporous resin (15 cm, φ1 cm, dry-loaded column), with

    water 100 ml, 30% methanol, 100 ml, methanol 100 ml elution, collecting methanol solution, and vacuum recovery to dry, and then fixed volume of methanol to 5 ml Liang Ping, namely, get the sample solution.

     2.3 The lack of hygiene drying parameters to take lack of preparation of solution samples of Sun-dried ginseng,

    according to the sample preparation method of solution made from lack of health-negative solution of drying parameters. Reposted elsewhere in the paper for free download http://

     2.4 column chromatographic conditions for the Kromail C18 column (4.6 mm × 150 mm, 5 μm), mobile phase of

    acetonitrile: 0.2% phosphoric acid (83:17), detection wavelength 203 nm, flow rate 1.0 ml / min , injection volume of 10 μl. In this chromatographic conditions, the taking ginsenoside Rg1 reference substance solution and test products

    for the solution, neither of Health and drying parameters negative reference substance solution of 10 μl, for HPLC

    determination of the results shown in Figure 1 ~ 3. The retention time consistent with the reference substance can be seen, and ginsenoside Rg1 and other components to achieve better separation. The negative does not appear that the peaks do not interfere with the determination of this element.

     2.5 Preparation of standard curve of ginsenoside Rg1 ginsenoside Rg1 were drawn precision reference substance

    solution (0.202 mg / ml) 2,4,8,10,15,20 μl injection,

    measured by the above method to sample volume abscissa, peak area value of the vertical axis, draw the working curve. Regression equation: Y = 122 262X 1 330.2, r = 0.999 1, r = 0.999 6, the linear range of 0.404 ~ 4.040 μg.

     2.6 precision experiment to continuously repeat the same reference substance solution into the sample five times to peak area of ginsenoside Rg1 of RSD = 0.23% (n = 5). The

results showed a good precision.

     2.7 to reproduce the experiment described by the former "determination" method, the same batch of samples, five copies were prepared for the test product solutions, each drawing 10 μl, for HPLC determination, the results of ginsenoside Rg1 content of 0.159 3 mg / g , RSD = 1.57%. Showed a good reproducibility of this Act.

     2.8 stability test to take the same test materials for the determination of solution according to method of injection measured at intervals of 2 h, a total of five times and the

    results showed that for the test items within the solution at 8 h basically stable, RSD is 0.27% (n = 5) .

     2.9 recovery rate increases are used as experimental sample recovery experiments, precise content of that is known to take samples (ginsenoside Rg1 content of 0.1593 mg / g), Precision adding a certain amount of reference substance, respectively, ginsenoside Rg1 reference substance solution (0.202 mg / ml) 4,5.5,6.5 ml were measured by HPLC sample were calculated recoveries, the average recovery was 96.34%, RSD of

    2.37% (n = 9). The results in Table 1. Recovery of experimental results in Table 1 (omitted)

     2.10 Determination of samples taken for the reference substance solution and test each product solution 10 μl, were

    injected into liquid chromatography, according to external standard method with peak area calculation, the determination of five batches of samples the results in Table 2. Table 2 Determination of ginsenoside Rg1 results (omitted)

     Based on the above determination results, tentatively

    scheduled for the test product containing ginsenoside Rg1 per g of not less than 0.15 mg.

     3 Discussion

     Experiments have used a variety of flow relative Wulongdan liquid extract component separation in the province, the results of acetonitrile: 0.2% phosphoric acid (83:17) as mobile phase, the peak-type ginsenoside Rg1 better separation

    effect is good, less interference of impurities, so use it as a mobile phase of this Law.

     Wulongdan liquid extract with water extraction method,

    more in line with clinical practice, but may affect the extraction rate of ginsenoside Rg1, therefore, the preparation of the extraction process depends on the clinical use of finalized. In this study, using HPLC determination Wulongdan Liquid Extract the content of ginsenoside Rg1 to prove that this method is simple, accurate, and reproducible recoveries were good.

     References

     [1] Hong Zhong, Xu-xiang. Wulongdan clinical

    application [J]. Journal of Modern Chinese Medicine, 2000, 9

    (4): 335.

     [2] Pheunkhang, Zang Kun Tong. Wulongdan the treatment of acute cerebral infarction and the clinical efficacy of transcranial color Doppler ultrasound analysis [J]. Chinese Journal of Emergency Medicine, 1999, 6 (9): 389.

     [3] Jian-Sheng Guo, Shen Qun, Wang Xiao-root, and so on.

    Wulongdan liquid extract quality standard [J]. Shizhen country Sinopharm Medicine, 2005,16 (12): 1273. Reposted elsewhere in the paper for free download http:/ /

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