Determination of ursolic acid in Dingli Jiangzhi capsule by TLC scanning

Objective: To study the content determination method of ursolic acid in Dingli Jiangzhi capsule: Method: thin layer scanning method was used to develop petroleum ether (30-60 ℃) - chloroform ethyl formate glacial acetic acid (9.7:0.8:1.8:0.16), with the determination wavelength of λ s = 540 nm and the reference wavelength of λ r = 420 nm: results: the linear range of ursolic acid was 0.24-1.20 g, R: 0.997 9 The recovery rate of sample addition is 97.0l%, RSD is 1.33% (n = 5) Conclusion: TLC scanning method is simple, rapid and accurate, which can be used for the quality control of the preparation: key words: Dingli Jiangzhi capsule; ursolic acid; content determination; TLC scanning method: Dingli Jiangzhi capsule consists of Dingli Zi, hawthorn and rhubarb They are composed of 7 kinds of Chinese herbal medicines, which have the functions of clearing and guiding stagnation, clearing collaterals and resolving stasis, and eliminating phlegm and dampness They are mainly used in the treatment of hyperlipidemia Hawthorn mainly contains organic acid, and the content of organic acid is mainly determined by neutralization method [Chinese Pharmacopoeia 2000] In order to control the internal quality of Dingli Jiangzhi capsule, the content of ursolic acid in hawthorn was determined by TLC scanning The result is accurate and the method is simple 1 Instrument and drug test cs930 thin layer scanner (Shimadzu company of Japan); silica gel G for chromatography (Qingdao Ocean Chemical Plant) Ursolic acid reference substance (China Institute of pharmaceutical and biological products, for content determination, batch number: 742-8902); Tingli Jiangzhi capsule (Jilin Tonghua Jinhui Pharmaceutical Co., Ltd., batch number: 010401-010410); all reagents are analytical pure 2 methods and results 2.1 test conditions: silica gel G plate (200 mm × 200 mm × 0.3 mm): Developer: petroleum ether (30-60 ℃) - chloroform ethyl formate glacial acetic acid (9.7:0.8:1.8:0.16), developer: 5% sulfuric acid ethanol solution, heated at 110 ℃ to purplish red, positioned under sunlight Scanning conditions: Take 1 μ l of ursolic acid reference solution (0.24 mg / ml), 1 μ l of test solution of Tingli Jiangzhi capsule, and take 1 μ l of negative sample solution prepared by the same method and place it on the same silica gel G plate, operate according to the law, and scan in the wavelength range of 370-700nm The results showed that Zui was greatly absorbed at 540 nm by both the reference and test solutions, while no interference was found in the negative sample solution Therefore, the measurement wavelength is set as λ s = 540nm, the reference wavelength is λ r = 420 nm, and the reflection sawtooth scanning is used The scanning speed is 10 mm / min and the slit is 1.25 mm × 1.25 mm 2 2 preparation of solution Test solution: take 6G of the content of this product, accurately weigh it, place it in Soxhlet extractor, add 60ml of ether, reflux for extraction for 5h, evaporate ether from the extraction solution to dryness, soak the residue in petroleum ether (30-60 ℃) for two times, each time for 15ml (about 2min), pour out petroleum ether, add anhydrous ethanol chloroform (3:2) solution to the residue to dissolve it, quantitatively transfer it to 5ml volumetric flask, and dilute it to the scale, Shake well, you will get it Control solution: accurately weigh 2, 4mg of ursolic acid control, place it in a 10ml measuring flask, add anhydrous ethanol to dissolve and dilute it to the scale Negative sample solution: take the other herbs except Hawthorn according to the prescription, make the negative sample of blood lipid reduction of Tingli according to the preparation process, take 6G powder, and prepare it according to the method under preparation of test solution 2.3 the linear relationship was investigated 1,2,3,4,5 μ l of the control solution was precisely absorbed, and then the control solution was placed on the same silica gel G thin layer plate, developed, colored and determined according to the law The regression equation is y = 54144.69x-2670.75 R = 0.9979 2.4 for stability test, take 2 μ l of ursolic acid reference solution, point the sample, unfold according to the law, air dry, spray with 5% sulfuric acid ethanol solution, bake at 105 ℃ for 4-7 min, cover the glass plate of the same size when it turns purple red, fix it, and measure once every half an hour until 3H Results the integral value of the absorbance of the control solution remained unchanged, RSD = 1.75% 2.5 precision test of the same plate: according to the law, point 5 samples of the same amount on the same thin plate, and point 2 μ L and 4 μ l each of the control solution Operate according to the law, measure the content of ursolic acid, and reflect the precision by the RSD value of the measurement result Results RSD = 1.20%, different plate precision test: on five silica gel G plates of the same size, point out the same amount of test sample respectively, and point at 2 μ L and 4 μ l of the control solution at the same time, operate according to the law, the content of ursolic acid was RSD = 1.35% 2.6 according to the reproducibility test, the same batch of samples were determined independently for 5 times The results showed that RSD = 1.44%: 2.7 sample addition recovery test took samples with batch number of 020713, the content of ursolic acid in the extract was 1.97 mg / ml, 5 samples were taken in total, 5 ml for each sample, and 5 ml (concentration of 200mg / ml) of the control solution were added precisely respectively According to the law, the recovery was calculated 2.8 for the negative interference test, take the above test sample solution, negative sample solution and control solution, separate and develop the color according to the above conditions, and then carry out the spectral scanning The negative sample solution had no absorption peak at the corresponding position of the test sample and the control sample solution, which indicated that other herbs except Hawthorn in the prescription did not interfere with the content determination of ursolic acid 29 The content of ursolic acid in the samples was determined by the method of determination of the sample content 2.10 the content of ursolic acid in Hawthorn is determined Take a proper amount of Hawthorn from each market, treat it according to the method under preparation of test solution, and determine the content of ursolic acid In order to ensure the quality of the product, it is necessary to limit the content of ursolic acid in Hawthorn to no less than 0.0l% before the dosage The method is simple and rapid, the results are accurate, the stability, precision and reproducibility are good, the recovery rate is ideal, it can be used for the quality control of Dingli Jiangzhi capsule.