Compound Yigan Tablets silybin by High Performance Liquid Chromatography
[Abstract] Objective To establish the compound Yiganling Tablets silybin method for the determination. Methods HPLC method, ODS C18 (4.6 mm * 250 mm, 5 μ m), with methanol - water - glacial acetic acid (48:52:1) as mobile phase, flow rate 1.0 ml * min-
1, detection wavelength 287 nm. Results silybin in the 13.75 ~ 110.00 μ g * ml-1 within
the concentration range of the sample with peak area showed good linear relationship
(r = 0.999 8), the average recovery was 97.10% (RSD = 0.63%, n = 6). Conclusion The method is simple, reliable, reproducible, and can be used in compound Yigan Tablets silybin analysis.
[Key words] high-performance liquid chromatography compound Yigan Tablets
Abstract: ObjectiveTo develop a method for determining the content of Silymarin in Fufang Yiganling tablets. MethodsHPLC analysis was carried out at the following conditions: Cromasil C18 column (4.6 mm * 250 mm, 5 μ m), mobile phase: Methanol-
Water-Acetic acid (48:52:1), the detection wavelength was at 287 nm the flow rate was 1.0 ml * min-1. ResultsThe standard curve was linear over the range of 5.9 ~ 29.5 μ g * ml-1 (r = 0.999 8), and the average recovery was 97.10% with RSD of 0.63% (n = 6);
ConclusionThis method is simple, reliable and reproducible, it can be used for the quantitative determination of the content of Silymarin in Fufang Yiganling tablets.
Key words: HPLC; Fufang Yiganling tablets; Silymarin
Compound Yigan Tablets with Yigan Zishen, detoxification Qushi effect. All side by silymarin, the five-jen alcohol extract composition, clinical for the liver and kidney, lead poisoning caused by damp outstanding Xie Tong, anorexia, abdominal distention,
lumbar acid, fatigue, Niaohuang embolism; or chronic hepatitis, transaminase levels were. The main components of drugs for the Compositae Silymarin Milk Thistle Silybum marianum (L.) Gaertn.'s Fruit, refined extract obtained by the loose powder,
the main ingredient containing flavonols, in which silybin has the strongest activity, and the a large extent, silymarin has the characteristics of the reasons, with heat-
clearing and detoxifying, Shugan Lidan achievements. The species is not Pharmacopoeia standards, ministerial standard method for the determination of specificity is not strong, are not high enough accuracy. ''2005 Edition of Chinese Pharmacopoeia ? Department already contains silybin method for the determination, the establishment of high-performance liquid chromatography compound Yiganling
Tablets Jun drug ingredient indications of the content of silybin is particularly necessary to . In this study, using HPLC Determination of Compound Yigan Tablets silybin content, with a view to a more comprehensive and effective control of the
An apparatus and materials
Shimadzu LC-10ATVP pump, SPD-10AVP UV detector; silibinin reference
substance (0856-200203, China's pharmaceutical and biological products); compound
Yiganling film produced by Jilin Aodong Taonan Pharmaceutical Co., Ltd. ; ultra-pure
water; methanol chromatography pure; acetic acid as the AR.
2 Methods and Results
2.1 The chromatographic conditions were column of ODS C18 (SHIMADZU, 4.6
mm * 250 mm, 5 μ m); mobile phase was methanol - water - glacial acetic acid
(48:52:1); detection wavelength 287 nm; flow rate of 1.0 ml * min-1 ; column
temperature was 35 ? .
2.2 Preparation of reference substance solution, said water fly thistle pin precise
amount of reference substance, add methanol made from milk thistle pin per milliliter of water solution of 0.05 mg, that is too.
2.3 Preparation of solution for the test items to take this product 5, except for sugar-coated, precision that set, and research Fine (over 100 mesh sieve), take 0.5 g, precision that set, and placing 150 ml bottle with cone-shaped plug, plus petroleum
ether (60 ~ 90 ? ) 50 ml, set the 90 ? water bath heated reflux 1 h, let cool, pour out
of petroleum ether, using dry filter paper filter, and then petroleum ether, 40 ml, sub-
sub-washed container residue and filter paper, filter Guo, Hui dry solvent, it will filter paper placed in the original cone-shaped bottle, precision plus methanol 50 ml, said the
fixed weight, set on a water bath heated reflux 1 h, put cold, weighing, filled with methanol, reduced weight loss, shaking uniform, filtration, abandoned to the beginning of the filtrate, precision continued to take the amount of filtrate 1 ml, set 10 ml Liang
Ping, add methanol was diluted to scale, shake, that is, too. Reposted elsewhere in the paper for free download http://
2.4 linear relationship between the study, said water fly thistle pin precision reference substance 13.75 mg, buy 50 ml Liang Ping, plus allows the dissolved and
diluted to the scale of methanol, shake. Each with precision drawing 0.5,1.0,2.0,3.0,4.0 ml to 10 ml flask, add methanol was diluted to scale, shake. And then 20 μ l were drawn into the precise liquid chromatograph, record chromatograms. In order to sample the
concentration of X-coordinate, in order to silybin and silybin different peak area and longitudinal coordinates for the linear regression, was the regression equation: Y = 1 017.40 10 376.02X, r = 0.999 8. Showed that silibinin in the 13.75 ~ 110.00 μ g * ml-1 of
the sample concentration range, with a good linear relationship between peak area. Linear regression curve shown in Figure 1.
2.5-negative sample interfere with inspection for water fly thistle pin reference
substance, compound Yigan Tablets for solution of test items, does not contain silymarin blank solution, measured according to injection, resulting in silibinin interference-free peak at retention time of impact.
2.6 precision experimental test products for precision drawing solution of 20 μ l, repeat sample six times, recording chromatograms result RSD = 0.24% (n = 6).
2.7 stability under the experimental precision obtained for the test product solution, 0,2,4,8 h has learned in 20 μ l injection of liquid chromatography to carry out the stability study, the results of RSD is 0.60%.
2.8 to reproduce the experiment to take the same group of samples were prepared according to the body of five copies of test items for the solution, under the same
chromatographic conditions, experimental results RSD of 0.33%.
2.9 recoveries experimental precision that is known to take six copies of the content of the sample powder 0.25 g, by adding the same amount of silybin reference
substance, according to the sample under assay method is to record chromatograms, computing the sample collection rate. The results in Table 1. The average recovery was 97.10%, RSD of 0.63% (n = 6). Recovery of experimental results in Table 1 (omitted)
2.10 Determination of content of the sample taken three batches of samples of 0.5 g, precision, said determined according to the law made for the solution of test items are based on 20 μ l sample were measured absorption peak area, other than the
standard method to calculate silybin content. The results in Table 2. Table 2 Sample assay results (omitted)
Different solvent systems have been selected as the mobile phase experiments, such as: methanol - water - glacial acetic acid (50:50:0.5), acetonitrile - methanol - water -
glacial acetic acid (5:45:50:0.5), methanol - water - ice acetic acid (48:52:1), etc. 
solvent system, the results of methanol - water - glacial acetic acid (48:52:1) separation
the best results. According to silybin UV absorption spectrum characteristic of silybin in the maximum absorption at 287 nm, choose 287 nm as detection wavelength.
This study established the compound Yigan Tablets silybin determination method for compound Yiganling tablets provide a basis for further improvement of the standard.
 State Pharmacopoeia Commission. Chinese Pharmacopoeia, a [S]. Beijing: Chemical Industry Press, 2005:56. Reposted elsewhere in the paper for free download