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High Performance Liquid Chromatography Injection in Shugan baicalin and Geniposide Content_3135

By Alice West,2014-10-30 19:29
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High Performance Liquid Chromatography Injection in Shugan baicalin and Geniposide Content_3135

    High Performance Liquid Chromatography Injection in Shugan baicalin and Geniposide Content

     Key words high-performance

     Abstract: Objective: To establish a high-performance

    liquid chromatography in Shugan Injection baicalin and

    Geniposide content. Methods: AtlantisTM dC18 column, 4.6 ×

    150mm, 5μ; baicalin mobile phase was 0.01mol / L phosphate buffer (pH2.8)-methanol (60:40), flow rate 1ml/min, detection wavelength 280nm; Gardenia glycosides mobile phase was methanol water glacial acetic acid (20:80:0.5), flow rate 1ml/min, detection wavelength 238nm. Results: baicalin and gardenoside linear in the range of 30 ~ 100μg/ml, 30 ~ 150μ

    g/ml, the average recoveries were 99.68%, 99.66%. Conclusion: The method is simple, rapid, accurate and suitable for Shugan

    Injection quality control.

     Key words: Shugan Injection; Determination; baicalin; geniposide

     Shugan Injection by capillaris, gardenia, Huang Qin, Isatidis and Ganoderma lucidum extracts made of sterile aqueous solution, attending Qingrejiedu, Lishi back yellow, Qi righting, nourish the liver Liver is an effective treatment for all kinds of viral hepatitis traditional Chinese medicine preparations, the main active ingredient of baicalin and gardenoside. There were determined by high performance liquid

    chromatography, or Geniposide baicalin content of the reports [1,2], but the sample column operation cycle is longer, in more than 15min. This study established in Shugan Injection baicalin and geniposide high performance liquid chromatography

    assay method with rapid, sensitive, simple, specific and strong characteristics of the active ingredient may be better to eliminate the interference between the .

     An instrument and reagent

     Waters High Performance Liquid Chromatography (including

    the 600 quaternary gradient pump, on-line Degasser, column

    temperature box, 996 diode array detector, Empower chromatography workstation), METTLER AE240 electronic analytical balance (Switzerland), METTLER 320 pH meter (Switzerland), cleaning En-Pu ultrasonic cleaning device

    (Shanghai), MS2 oscillator (Germany), PIERCE dry and hot-type

    sample concentrator (USA), Millipore ultrapure water device. Baicalin and gardenoside reference substance from China provided medicines and biological products. Methanol for

    chromatography pure reagents (U.S. TEDIA company), other reagents were analytical pure. Shugan Injection Pharmaceutical Co., Ltd. of Guizhou Shui production (batch, respectively 0409091,0411052,0503301).

     2 Methods and Results

     21 chromatographic conditions

     AtlantisTM dC18 column, 4.6 × 150mm, 5μm; baicalin

    mobile phase was 0.01mol / L phosphate buffer (pH2.8)-methanol

    (60:40), flow rate 1ml/min, detection wavelength 280nm; geniposide flow phase, methanol water glacial acetic acid

    (20:80:0.5), flow rate 1ml/min, detection wavelength 238nm.

     22 Preparation of Solution

     221 Preparation of reference substance solution

     Precision Weigh baicalin and gardenoside amount of the reference substance, dissolved amount of methanol, water will dissolve, respectively, made 30,45,60,75,100 μg / ml baicalin

    reference substance solution, 30,60,90 , 120,150 μg / ml

    solution of Geniposide reference substance.

     222 Preparation of sample solution

     Precise volume of test items taken for the solution of 2mL to 25mL Liang Ping, the methanol was diluted to scale, shake, using 0.45μm microporous membrane filter, filtrate, as measured geniposide sample solution. Re-learn the precise 2mL

    filtrate home 25mL Liang Ping, the use of methanol diluted to

    scale, shake, as the determination of baicalin in sample solution. Reposted elsewhere in the paper for free download http://

     Were collected from baicalin and gardenoside reference substance and sample solutions 10μL, in the above

    chromatographic conditions, sample injection, was the above-

    mentioned two points for the reference substance and the test solution chromatogram of products, the results shown in Figure 1,2.

     23 linear relationship between the study

     Under the chromatographic conditions in the above two components followed by a series of standard solutions measured by the concentration of reference substances as abscissa and peak area of the vertical axis linear regression analysis. Baicalin and geniposide regression equation were A = 3.26 ×

    10-7C-0.01, r = 0.9999; A = 7.04 × 10-7C-0.002, r = 0.9998.

    The results showed that baicalin at 30 ~ 100μg/ml, within the

    30 ~ 150μg/ml Geniposide showed good linear relationship within.

     24 precision test

     Determination of sample solution by using the same

    chromatographic conditions, repeated injection of sample solution (n = 6), measured peak area, calculated baicalin and geniposide the relative standard deviation (RSD) were 0.98% and 1.04%.

     25 Stability Test

     Take 2.4, respectively, 0,2,4,8,12 h under the solution

    into the sample, measured peak area, calculated RSD. The results showed that the sample was stable in the 12h, baicalin and geniposide RSD were 0.98% and 1.05%.

     26 recovery test

     Sample collection method of adding a known concentration

    of the Injection in Shugan, respectively, by adding different amounts of baicalin and gardenoside reference substance, according to 2.2.2 under the method of operation, may recovery

experimental solution, taking 10μL sample, calculate the

    recovery rate, the results in Table 1. Table 1 Sample recovery (omitted)

     Reference standards and samples were collected from solution, depicting 10μL sample, using external standard

    method to calculate content of the sample results in Table 2.

    Table 2 Determination of Sample Results (abbreviated)

     3 Discussion

     Shugan Injection of main active ingredients of baicalin and gardenoside, so their quantitative, clearly the scope of its content is to control the quality of the key. Baicalin and

    geniposide physical and chemical properties and UV absorption characteristics are quite different, it is difficult to obtain in conventional conditions, at the same time separated to meet the quantitative requirements. Long-Lin [1,2] reported the

    high-performance liquid chromatography of their baicalin and geniposide content, but the sample on the column to run longer. This study established UV detection at two wavelengths, using two kinds of mobile phase were measured by baicalin and geniposide content method with rapid, sensitive, simple, specific and strong characteristics that can be better to eliminate the effective interference among the components suitable for Shugan Injection quality control.

     References

     1-Long Lin, Zhang Yun, Huang Shunju. HPLC Shugan

    Injection Determination of baicalin content. Chinese medicine, 2003,25 (12): 1025

     2-Long Lin, Zhang Yun, Huang Shunju. HPLC determination of Shugan Injection Geniposide content. Chinese medicine, 2003,25 (10): 852 reposted elsewhere in the paper for free

    download http://

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