Asthma Ning Granules Determination of baicalin
【Abstract】 Objective To establish the asthma baicalin
Granule method for the determination to develop a quantitative quality control standards for the preparation. Methods of
methanol 4% phosphoric acid solution (55:45) as mobile phase, detection wavelength was 280 nm. The results of baicalin measured linear range of 12.25 ~ 125.2 μg / ml, r =
0.999 9, the average recovery was 98.8%, RSD 1.43%. Conclusion of this preparation in the determination of baicalin in
approach, the method is accurate, sensitive, specific, reproducible, no significant interference, etc., is to control the inherent quality of the goods, the more ideal way.
Key words asthma Granule performance liquid chromatography of
Asthma Granule is a skullcap, ginkgo, land dragon, Morus alba L. composed of 10 Chinese herbal medicines Chinese herbal compound preparation of Chinese medicine for the new drugs are being developed, the side with a heat asthma, relieving cough
and phlegm of the efficacy of the bronchial asthma, asthmatic bronchial asthma, emphysema and other respiratory diseases with good results. Prescriptions Jun Scutellaria based medicine, in order to control the product's inherent quality,
and develop quantitative quality control standards, reference
to relevant literature [1 ~ 4], according to Chinese medicine as declared by the technical requirements, study and formulate a method for the determination of the drug . Are reported
An instrument and reagent
High-performance liquid chromatography (pump: the United States Waters 510, Waters 2487 detector). Column for the Japanese company Shimadzu shim pack CLC ODS, 6.0 mm ×
150 mm, ultra-pure water (millipore's Water Purification System), used for the chromatography of pure methanol, and
other reagents were AR, baicalin control goods purchased in China's pharmaceutical and biological products for the determination of use, lot number 0715 9909.
2 Methods and Results
2.1 Preparation of sample solution
2.1.1 Preparation for the test product solution obtained under the load difference amount of this product, and research fine, take 0.5 g, precision that set, plus 70% ethanol, 40 ml, refluxed 2 h, put cold, filtration, the filtrate Purchase 100
ml Liang Ping, the 70% ethanol, washed with a small amount of residue on the filter paper and Liang Ping, add 70% ethanol to scale, shake, as a solution for the test items.
2.1.2 Preparation of reference substance solution, vacuum dried at 60 ? taken 4 h the amount of baicalin reference substance in 50 ml Liang Ping, the precision that as 15.65 mg, plus allows the dissolved methanol, and diluted with methanol to scale, shaking uniform, as a reference substance stock solution, its concentration of 0.313 mg / ml; precise amount of take reference substance stock solution 2 ml, set 10 ml Liang Ping, add methanol to the scale, shake, as a reference substance solution, its concentration of 62.6 μg / ml.
2.1.3 Solution Preparation medicinal Scutellaria
Scutellaria Radix obtained powder 0.3 g, precision that set, plus 70% ethanol, 40 ml, refluxed 3 h, put cold filtration, the filtrate 100 ml Liangping in the home, using a small amount of 70% ethanol washed filter paper and residue, plus 70% ethanol to scale, shake, precision volume Liangping take 1 ~ 10 ml, add 70% ethanol to scale, shake, as a medicinal Scutellaria solution.
2.1.4 lack of preparation of Scutellaria blank solution and accurately according to the proportion of prescriptions
taken except that skullcap taste than other herbs to simulate the preparation of this product technology and product solutions for the test preparation methods, made of skullcap missing blank solution.
2.2 Chromatographic conditions and system suitability
test Column: The 18 alkyl silane bonded silica gel as a column filler (Shimadzu Shim pack CLC ODS column, 6.0 mm × 150
mm); methanol 0.4% phosphoric acid solution (55:45) as mobile phase; flow rate of 1.0 ml / min; detection wavelength
280 nm; temperature is room temperature; theoretical plate number calculated by baicalin not less than 2 000.
2.3.1 The maximum absorption wavelength of the alternatives are sophisticated drawing reference substance solution and test products for the solution of 10 μl, into
the liquid chromatograph, in the range of 200 ~ 400 nm to detect. The results showed that reference substance solution for the test materials are in solution at 280 nm have the same maximum absorption peak detection wavelength was therefore determined to be 280 nm.
2.3.2 interference experiment, precision drawing a blank reference substance solution and test products for the solution and the lack of Scutellaria blank solution 10 μl,
into the liquid chromatograph, and detected at 280 nm showed that baicalin reference substance solution and for the Test product solution at the wavelength of maximum absorption peaks are rather lacking in the blank solution Scutellaria wavelength no obvious peaks appear (see Figure 1), indicating
that no significant interference in this determination.
2.3.3 linear relationship between the inspection precision drawing solution of the above-mentioned reference
substance reserve amount, with methanol concentrations were
prepared as 12.52,31.3,62.6,93.9,125.2 μg / ml solution,
precision drawing reference substance solution of the above 10 μl, injection efficiency liquid chromatography to measure peak area values, the results in Table 2. To peak area values (A) for the vertical axis, baicalin sample volume (μg / ml)
as abscissa, drawing standard curve, the results of sample volume of 12.25 ~ 125.2 μg / ml range a good linear
relationship, on income data, regression analysis was the regression equation y = 110 884x-28 837, the correlation
coefficient r = 0.999 9. Linear relationship between the study results in Table 1 (omitted) reposted elsewhere in the paper for free download http://
2.3.4 precision experimental test products for precision drawing solution 10 μl, sample 5 consecutive times, measured peak area and calculating the precision, the results for the test items measured five times, RSD was 1.15%, indicating good precision.
2.3.5 Stability of test items for the experimental precision drawing solution of 10 μl, at intervals of 2 h into
the determination of a sub-sample, sample measured five times
together, its peak area values in the determination of time, basically unchanged, within 8 h for the test materials The RSD for five determinations of 2.25%, indicating good stability.
2.3.6 repeated experiment to take the same group of samples (20,050,807) 6 copies, according to determination under the law, determine the content of baicalin, the results of six determinations, the average content of 17.15 mg / g,
the RSD is 1.52%, indicating that reproducibility was good. 2.3.7 recovery experiment to take the same group of samples (20,050,807), taking a total of five copies of baicalin were
added 4.05 mg (dubbed in the concentration of 1.35 mg / ml solution, accurate and adding 3 ml), according to the above samples were determined the content of baicalin and calculate the recovery rate, the results of five times the measured
average recovery was 98.82%, RSD 1.43%. The results in Table 2. Recovery of experimental results in Table 2 (omitted)
2.3.8 Sample measurement precision drawing reference substance were awarded for the test solution and three different product solution 10 μl, into the liquid
chromatograph to measure and calculate the various batches of samples the content of baicalin. The results in Table 3. Table 33 batches of samples in the determination of baicalin in results (omitted)
2.3.9 three groups Scutellaria Radix determination of baicalin by "Chinese Pharmacopoeia" 2005 Edition (?
Division), p. 211 baicalin baicalin-containing measuring
method three groups were measured herbs Scutellaria results of batch baicalin baicalin in Radix contents are in line with
"Chinese Pharmacopoeia" 2005 Edition (? Department) on the
content of baicalin not be less than 8.0% of the requirements (used in this study baicalin baicalin content of 11.0%). The determination of the results of all batches of samples in Table 4. Table 43 groups Scutellaria Radix Determination of baicalin results (omitted)
Methodology of the system through the investigation and determination of the three batches of samples was established asthma Granule baicalin in HPLC assay method, measured by
three batches of samples of the contents of baicalin were 17.93,16.32,17.06 mg / g, the sample recovery rate was 98.82%, RSD 1.43%.
The determination of the solution in preparation for the test materials, using 70% ethanol extract of the direct point-
like determination, the method is simple, accurate, high sensitivity, specificity, good reproducibility, no significant interference, etc., is to control the the intrinsic quality of products ideal way. As well as other preparations can be used
as reference for determination of the same composition.
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