High Performance Liquid Chromatography expectorant cough mixture in the content of baicalin
Author: Fuling Yan Xiao-Ying Cheng Zhao will be
【Abstract】 Objective: To establish expectorant cough mixture method for the determination. Methods: High
performance liquid chromatography (HPLC) Determination of expectorant cough agent content of baicalin. Results: HPLC method measured the product of the content of baicalin 0.512 ~ 0.541mg/ml, in the range of 0.40 ~ 2.00μg, the solution
concentration and peak area showed good linear relationship, r = 0.9993, plus the average sample recovery was 99.85% (n = 5), RSD 0.69%. Conclusion: This product is quantitative method is simple, accurate, specific and able to control the inherent
quality of the preparation.
Key words expectorant cough agent high-performance liquid
chromatography determination of baicalin
Determination of Baicalin in Qutanzhike Mixture by HPLC
【Abstract Objective】 Study on the Quality Standarded
for Qutanzhike Mixture. Method: Radix Scutellariae, Flos Lonicerae in Qutanzhike Mixture were identified by TLC, and the content of Baicalin was determined by HPLC. Result: The Baicalin showed a good linear relationship within a range of 0.40 ~ 2.00ug (r = 0.9993), the average recovery was 99.96% (n = 5), RSD = 1.37%. Conclusion: method HPLC is accurate and reliable method.The quality standard can be used for quality control of this product .
【Key words】 Qutanzhike Mixture HPLC Baicalin
Expectorant cough mixture from the skullcap, bitter almond, licorice, drugs such as the composition of Trichosanthes Jen. The party is on the basis of a large number of clinical practice in order to TCM-based, combined with
modern medicine and modern pharmacological developed the
treatment of acute and chronic bronchitis effective drug. In order to better control its quality, and ensure that the preparation of clinical efficacy, this study used HPLC method for preparation of the main active ingredient content of
baicalin were determined, are reported below.
An experimental material
1.1 Instrument LC-2010A high-performance liquid
chromatography, CLASS-VP chromatography workstation, UV-2401
UV spectrophotometer (Shimadzu, Japan).
1.2 reagent baicalin reference substance (China Pharmaceutical and Biological Products Institution, for the determination of use, Lot :110715-200212); expectorant cough
mixture (Jilin Chuanying Qu Chinese medicine hospital preparation room); methanol chromatography pure ; the other
reagents were analytical pure. Reposted elsewhere in the paper for free download http://
2 Methods and Results
2.1 Chromatographic conditions and system suitability test column: DiamosilTM (diamonds) C18 (5μ 250 × 4.6mm,);
mobile phase: methanol - water - glacial acetic acid
(45:55:1); detection wavelength: 276nm; flow rate: 1.0ml/min; column temperature: room temperature (25 ?); injection
volume: reference substance solution 6μl, sample solution 10
μl; theoretical plate number by baicalin peak meter should not be less than 1500.
2.2 Preparation of Solution
2.2.1 Preparation of reference substance solution, said
precision vacuum drying at 60 ? to take 4h amount of baicalin
reference substance, add methanol is made with 0.1mg per 1ml
of the solution, as a reference substance solution.
2.2.2 Preparation for the test solution precision the amount of goods to take this product 10ml, home 50ml flask, add 60% of the amount of methanol, ultrasonic (power 250W) for 20 minutes, place to room temperature, plus 60% methanol to scale, shaking uniform, as a solution for the test items.
2.2.3 Preparation of negative control solution, to take except skullcap rest of prescription medicines other than the one-tenth of the amount, according to the rule of law into the mixture, and then test products for the preparation, made from the negative control solution.
2.3 Specificity Test
Precision for the test items were drawn solution, negative control solution, 10μl, reference substance solution
6μl, into the liquid chromatograph, record chromatograms (Figure 1). Can be seen from the figure for the test materials
chromatography, chromatography with the reference substance for the corresponding position, have the same retention time (30.8min) of the chromatographic peaks, the negative test without interference, to prove feasibility of this Act.
2.4 linear relationship between the study said that taking precise amount of baicalin reference substance, add methanol produced contains about 0.1mg per 1ml of solution, respectively, to take precise amount of reference substance solution 4,8,12,16,20 μl, into the kind of determination in order to concentration (X) as the abscissa and peak area (Y) for the longitudinal coordinates, was the regression equation Y = 1151340.8 100157.4 X, r = 0.9993, linear range of 0.40 ~ 2.00μg.
2.5 precision and stability testing to take samples (batch number: 050,517) according to "2.2.2 Preparation for test materials solution" approach to preparation of the sample solution, repeat sampling five times sample volume 10μl, its
relative standard deviation of peak area (RSD) of 0.9%, indicating good precision. To take the same sample solution, in the 0,2,4,10,24 h sample were measured, the results of five
Times the measured value (baicalin, mg / ml), RSD 1.3%,
showing that baicalin was stable in the 24h.
2.6 to reproduce the test to take for the test product (Lot No.: 050,517), a total of five copies, respectively, "2.2.2 Preparation for test materials solution" approach to preparation of sample solution, were measured, the RSD of baicalin content of 0.7 %, indicating good reproducibility.
2.7 Precision tests the amount of recoveries to take a known concentration of sample (Batch No: 050,516, content
0.529mg/ml) amount, a total of five copies, respectively, by adding quantitative amount of baicalin reference substance, according to "2.2.2 for test Preparation of product solution "approach to operations, may recovery test solution, according
to the measurement results in Table 1.
2.8 Determination of the tested products and limits the development to take several groups of the tested products, according to the methods described in the body of the content determination of baicalin, the results of all batches of samples of the contents of baicalin were higher than 0.50mg/ml, tentatively based Each 1ml contains items to baicalin baicalin (C21H18O11) dollars, shall not be less than 0.40mg. Table 23 Assay results of samples
3.1 Determination of detection wavelength detection wavelengths in the choice, I take samples in the 200 ~ 350nm wavelength range scanned, resulting in maximum absorption at 276.5nm, so choose for the detection of wavelength 276nm.
3.2 The author compared the choice of mobile phase of methanol - water - phosphoric acid (50:50:0.3), and methanol -
water - glacial acetic acid (50:50:1), etc. The mobile phase, the results of methanol - water - glacial acetic acid
( 45:55:1) separation of the best results.
HPLC method used in the preparation of baicalin quantitative, accurate, convenient simple, reproducible, and recoveries were satisfactory, you can effectively control the product's inherent quality.
 National Pharmacopoeia Committees. The People's Republic of China Pharmacopoeia [M]. 2005 Edition, 1. Beijing: Chemical Industry Press, 2005,1,370 ~ 371,405 ~ 406. Reposted elsewhere in the paper for free download http://