Guanxin plain film method of identification and determination of
【Abstract】 Objective To establish a coronary plain film of the identification and determination methods. Thin Layer Chromatography (TLC) method of preparation of the Chinese
angelica, Trichosanthes skin, 37 are identified, and preparations by ultraviolet spectrophotometry of total polysaccharide content. Chromatographic characteristics of the results of TLC spots obvious. Measured by three batches of samples, the polysaccharide content is 14.76% the highest, the lowest was 12.24%, with an average of 13.36%; The average recovery was 97.38%, RSD of 5.51%. Conclusion thin-layer
chromatography (TLC) method for the preparation of the Chinese angelica, Trichosanthes skin, 37 to identify and ultraviolet
spectrophotometry method of determination of total polysaccharide simple, accurate, specific, plain films can be used as coronary heart quality control methods.
Key words coronary plain film; polysaccharide; TLC; UV spectrophotometry
Abstract: ObjectiveTo establish quality standards for Guanxinping Tablet. MethodsRadix Angelicae Sinensis, Pericarpium Trichosanthis and Radix Notoginseng were identified by TLC, and the content of polysaccharides was determined by UV spectro. Photometry. ResultsThe TLC spots developed were quite clear. The hightest content was 14.76%, the lowest content was 12.24%, the average content of polysaccharides was 13.36%. The average recouery of polysaccharides was 97.38%, with RSD of 5.51%, respectively.
ConclusionThe method is convenient, reliable. It is one of the ways that can be used for the quality control of Guanxinping tablet.
Key words: Guanxinping tablet; Polysaccharides; TLC; UV
Guan Xin-Ping film by Wong Jing, Chinese angelica,
Trichosanthes skin, composed of 37 traditional Chinese medicine, etc., with qi, nourishing yin and promoting blood circulation Huayu Qi, Shu by Tongmai pain relief. Qi Yin is mainly used for clinical blood loss injuries, phlegm
congestion gas block, shortness of breath, fatigue, heart palpitations embolism, and arrhythmia, atherosclerosis, high blood lipids, etc.. In order to ensure quality of preparation, the author carried out the preparation method of identification and determination. Now report on the results as follows.
An instrument and reagent
WD 9413A gel imaging analyzer: Beijing Instrument Factory 61 products; electronic analytical balance, BP211D Ohaus (Shanghai) Corporation; TU 1800S UV-Vis
spectrophotometer, Beijing Pu An Analysis of General Instrument Co., Ltd. products; 939 thin-layer planking
machine, Chongqing shellfish available instrumentation products.
1.2 Materials and reagents
Silica gel G, silica gel GF254, Qingdao Nanyang Chemical
Corporation products; silica gel G plate, silica gel GF254 plates, self-made; coronary plain film, Jiangsu Province, Department of Medicine Hospital agent production, batch number 020812,041111,050522; anhydrous glucose , Shanghai chemical
reagent procurement and supply station Reagent Factory production; the reagents were analytical pure.
2 Methods and Results
2.1.1 37 TLC method
To take this product 10, and research fine, plus ethanol, 50 ml, refluxed 1 h, filtration, filtrate water bath, evaporated and the residue dissolved in 20 ml water, using n-
butanol 40 ml, 2 times, extraction, combined n-butanol fluid,
washed two times with ammonia test solution, 20 ml / times, disposable ammonia test solution, n-butanol liquid evaporated
and the residue add 2 ml of methanol allows the dissolved, as a solution for the test items; and then take 37 medicines 2.5 g in control with the rule of law must control 37 medicinal solution, as a reference substance solution; an alternative
non-negative samples made of 37 herbs 10, the same treatment, made negative reference substance solution. According to thin-
layer chromatography experiments, learn from the above-
mentioned three kinds of solution, the 10 μl, respectively,
points on the board with a silica gel G, ethyl acetate to chloroform methanol-water (15:40:22:10 ) 10 ? below
the lower placement solution as the mobile phase, to start out, dried. Sprayed with 10% sulfuric acid ethanol solution, heated at 105 ? for spot color definition. Test products for chromatography, in the corresponding position with the reference substance chromatography, significantly the same color of the spots, no such negative control spots. Figure 1.
2.1.2 Angelica TLC method
To take this product 20, and research fine, plus 1% sodium carbonate solution 50 ml, ultrasonic extraction 30 min, centrifuged and the supernatant, add hydrochloric acid pH adjusted to 1 ~ 2, using ethyl ether 60 ml, divided into 3 sub-extraction The combined ether solution, and washing three times, 20 ml / times, abandoned the water liquid, ether liquid evaporated and the residue add 1 ml of methanol allows the dissolved, as a solution for the test items; and then take control angelica herbs, with the rule of law into a reference substance solution. An alternative non-negative samples made
of Angelica sinensis 20, with the rule of law into a negative reference substance solution. According to thin-layer
chromatography test, drawing the above-mentioned three kinds
of solution of 5 μl, respectively, points on the board with a silica gel G, ethyl benzene acid (4:1:0.2) as mobile
phase to start , removed dry. Purchase UV (365 nm) under review. Test products for chromatography, in the corresponding
position with the reference substance chromatography, significantly the same color of the fluorescent spots, negative control no such spots. Figure 2.
2.1.3 Paper TLC method of Trichosanthes
To take this product 10, and research fine, plus ethanol,
50 ml, refluxed 1 h, filtration, filtrate water bath, evaporated and the residue dissolved in 20 ml water, with ether 60 ml, divided into 3 sub-extraction, 20 ml / second.
Combined ether solution, and washing three times, 20 ml / times, abandoned water liquid, ether liquid evaporated and the residue add 1 ml of methanol allows the dissolved, as a solution for the test items; and then take control medicinal Trichosanthes Paper 2 g, with the same The method for producing reference substance solution; an alternative non-
medicinal Trichosanthes skin made of 10 negative samples, with treatment, made negative reference substance solution. According to thin-layer chromatography experiments, learn from the above-mentioned three kinds of solution of the 5 ~ 10 μl,
respectively, points on the board with a silica gel G, petroleum ether (60 ~ 90 ?) ethyl acetate (4:1) as Expand
agent, started out, dried, sprayed with 5% phosphomolybdic acid test solution, heated to 105 ? in a clear color. Test
products for chromatography, in the corresponding position with the reference substance chromatography, significantly the same color of the spots, no such negative control spots. Figure 3.
Huang Jing, as a kind of Bu Zhong Yi Qi, kidney Tianjing,
Ziyin lungs, Sheng Jin Spleen efficacy of traditional Chinese medicine, on the treatment of cardiovascular diseases, tuberculosis, chronic hepatitis, as well as in anti-bacterial,
detoxification, anti-fatigue, anti-aging, etc. involved will
have a good function, its active ingredients is mainly Polygonatum polysaccharide. During the Fang, to fill the heart of Polygonatum gas-run power of the elected Jun
cardiopulmonary medicine, so I chose the active ingredient in Polygonatum Polygonatum polysaccharide as the main objective
of [a] determination of total polysaccharide preparations.
According to spectrophotometric determination of total polysaccharide content . Reposted elsewhere in the paper for free download http://
2.2.1 Preparation of reference substance solution
Precision Weigh 105 ? drying to constant weight of
glucose reference substance 36.3 mg, buy 100 ml Liang Ping, add water, dissolve and dilute to the scale, shake, or have (each ml containing anhydrous glucose 0.363 mg).
2.2.2 Determination of Wavelength Selection
Precision drawing reference substance solution, 0.1 ml, set 10 ml scale in a test tube with plug, add water to 2.0 ml shaken, the ice bath slowly dropping 0.2% anthrone - sulfuric
acid solution to scale, mixing, post-release cold water bath
in the insulation 10 min, removed and immediately cooled in ice-bath home 10 min, removed the corresponding reagent blank, at 400 ~ 700 nm wavelength range scanning, mapping absorption spectra, results showed that glucose in the maximum absorption
at 528 nm.
2.2.3 Preparation of standard curve
Precise amount of reference substance solution to take 0.1,0.2, 0.3,0.4,0.5, 0.6 ml each purchase 10 ml test tube with plug scale, each adding water to 2.0 ml shaken, the ice bath slowly dropping 0.2% anthrone - sulfuric acid solution to
scale, mixing and let the cold water bath in the warm post-10
min, removed and immediately cooled in ice-bath home 10 min,
removed the corresponding reagent blank. According to UV -
spectrophotometry at 528 nm wavelength measured absorbance to the absorbance of the vertical axis, the concentration of abscissa, drawing standard curve. Figure 4.
2.3 Stability test
Precision drawing reference substance solution, 0.3 ml, set in 10 ml Liang Ping, according to standard operating under
the curve were measured at 0,10,20,30,40 min after the preparation absorption. The results show that absorption within 40 min remained unchanged (RSD 0.32%), the maximum absorption wavelength is also unchanged.
2.4 Precision Experiment
Precision drawing reference substance solution, 0.3 ml, a total of six copies, respectively, 10 ml Liangping in the home, according to the standard curve of absorbance measured under the operation, RSD is 0.32% (n = 6).
2.5 The average recovery experiment
Take a known concentration of the finished product (batch number 041,111) 2, research small, precision that set, plus 80% ethanol 100 ml, heated reflux 1 h, filtration, residues with 80% ethanol, washed three times, and then precision Join
105 ? drying to constant weight after the glucose reference substance, add 100 ml water, reflux extraction 1 h, filtration, the filtrate 200 ml flask in the home, add water to the scale, shake, sophisticated take this amount of liquid
0.2 ml, according to "standard Preparation of the curve, "under the" all adding water to 2.0 ml shaken, the ice bath slowly dropping 0.2% anthrone - sulfuric acid solution to 10
ml scale, "from operations, according to the measured absorbance. The results in Table 1. Recovery of experimental results in Table 1 (omitted)
2.6 Determination of Sample
To take this product 4, and research detailed, precise, said determined set Round-bottom flask, add 80% ethanol 150
ml, set in a water bath reflux 1 h, while hot filtration, residue washed with 80% hot ethanol three times, 10 ml / times, will be set residue and filter paper in the flask, add water, 150 ml, set in boiling water bath heating reflux 1 h, while hot filtration, residue and the flask washed with hot water four times, 10 ml / times, combined filtrate and lotion, put cold, transferred to 250 ml Liang Ping, add water to
scale, shake, take precise amount of 0.1 ml, set 10 ml dry test tube with plug, according to the "standard curve prepared" under the "all adding water to 2.0 ml shaken, the ice bath slowly dropping 0.2% anthrone - sulfuric acid
solution to 10 ml scale, "from operations, according to the measured absorbance to calculate content of the sample. The results in Table 2. Table 2 Determination results of Guan Xin-
Ping polysaccharide (omitted)
3.1 using TLC method Angelica, 37, Trichosanthes skin to TLC, in order to medicines control, simple, rapid, repeatable, high-specific advantages. Angelica doing the identification, the product of adding 80% ethanol was extracted water bath, filter, add diluted hydrochloric acid pH adjusted to 1 ~ 2,
extracted with ether three times, combined ether solution, evaporated and the residue plus methanol 1 ml, As for the test items allows the dissolved solution with benzene - methanol -
acetic acid (30:3:1) as the agent, started out, drying, home
UV lamp (254 nm), under review for the test materials chromatography and Reference Substances poor chromatographic separation. In 37 identification, press the "Chinese Pharmacopoeia" approach with chloroform methanol water
(13:7:2) 10 ? below the lower placement solution as the mobile phase, but when there is no stratification in the preparation, sample separation is not clear, the latter replaced by chloroform, methanol ethyl acetate-formic
acid (40:20:10:0.2) as the agent for 10% of the sulfuric acid
ethanol as chromogenic reagent, the color spot is not clear, good separation. At last, chloroform methanol water,
ethyl acetate (15:40:22:10) 10 ? below the lower placement
solution as the mobile phase, to start out, dried. Sprayed
with 10% sulfuric acid ethanol solution, heated at 105 ? for
a clear spot color separation effect is good.
3.2 Guanxin plain film group Fang, Huang Jing-based
medicine, Polygonatum polysaccharide is one of its active ingredients, refer to "Chinese Pharmacopoeia" Huang Jing
medicinal use colorimetry to measure under Polygonatum polysaccharide with a simple, accurate, good reproducibility and can be used as a quantitative coronary plain film detection method. However, Angelica, 37, Trichosanthes bark
polysaccharides is also the active ingredient, according to the sample preparation process of the negative reference substance missing Polygonatum at 528 nm Department has absorbed. Therefore, the use spectrophotometry in which the total content of polysaccharides.
Content of indicators to determine: three batches of samples determined by analyzing the results of modest content changes, the content up to 14.76%, the lowest of 12.24%, with an average of 13.16%, taking into account the current source
and the quality of herbs and other factors of instability are more temporary taking the mean of the 2 / 3 as the lower limit, that is, the polysaccharide content of finished 9.0% lower limit.
3.3 recoveries, since the glucose reference substance can be dissolved in 80% ethanol, so when the extracted sample was extracted with 80% ethanol, after, and then glucose added to the samples after treatment and then recovery experiments .
3.4 conducting recovery experiments, RSD = 5.51% (n = 6), a larger error. The reason is that the stability of the method is shorter, by adding anthrone reagent after only 40 min to stabilize, anthrone easily decomposed when exposed to light, the operation should be joining the anthrone, and rapidly determined.
 Pang Xin, Zhi Zhao, Yuan Yuan. Polygonatum the chemical composition and pharmacological effects [J]. Mountain Agriculture and Biology, 2003,22 (6): 547.
 National Pharmacopoeia Committee. Chinese
Pharmacopoeia, ? Department of [S]. Beijing: Chemical
Industry Press, 2005:215, Appendix VIB. Reposted elsewhere in the paper for free download http://