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Extraction of active ingredients of saffron Zhiyuan Medical Papers Online: www.zykkk.cn journal writing medical papers published translation changes Tel: 13871398507 QQ: 278717711 E-mail: email@example.com Key words HPLC method Abstract: Objective: To screen the active ingredient in the extraction process of safflower. Methods: RPHPLC determination of safflower extract HSYA A as quantitative indicators, and the uniform application of single-factor experimental design, selection extraction safflower active ingredients. Results: The extraction of water as solvent, reflux 2 times, each time 20 min, each time the amount of water to 13 times the amount of medicine. Per gram of safflower extract herbs HSYA A was the highest. Conclusion: The extraction process is stable and feasible. Keywords: Safflower; active ingredient; RPHPLC method; HSYA A To studies of extraction methods of utility portion in Carthamus tinctorius YANG Xiaojun, WU Guirong, WANG Yan (College of Pharmacy, Xinjiang Medical University, Urumqi, 830054 China) Abstract: Objective: To screen the extracting method of utility portion in Carthamus tinctorius. Methods: A RPHPLC method was developed to determinate the content of hydroxysafflor yellow A in extracting portin of safflor yellow. According to the mono factor design and uniform design. We screened the extracting method of utility portion in Carthamus tinctorius. Results: The extracting solvent was water, repeat 2 times, every time add 13 times water and the extracting lasts 20 min to the Chinese medicine drug. The content of hydroxysafflor yellow A was highest in extraction for every gram Carthamus tinctorius. Conclusion: The method is simple, stabilization and feasible. Key words: Carthamus tinctorius; utility portion; RPHPLC; hydroxysafflor yellow A Safflower (Carthmus tinctorins L.) is a dry tubular flowers of Compositae , only contained in the "Kai Bao Materia Medica," "Lord of postpartum blood supply Koujin, not the bad blood intra-abdominal, cramps, abdominal fetal death in, and Jiuzhu service, also in Gudu of blood. " With the expansion of its coronary artery, blood pressure, hypoxia, blood circulation, anti-inflammatory analgesic and other pharmacological effects . Germany and Japan in 1906, high-level turtles first was isolated from the red pigment in safflower (C12H22O11) content of 0.3% to 0.6%; yellow pigment mixture containing a variety of chalcones ketones (Safflor yellow, SY), the content of 20% to 30 %. Pharmacological experiments confirmed that  SY is the main active ingredients of effective, SY hydroxyl yellow A (hydroxysafflor yellow A, HSYA) were higher, with a typical pharmacological effects . In the land rich in natural resources, plenty of sunshine, the production of all-natural, pollution-free good base of green herbs, safflower production accounting for about 80%. In this study, water as solvent extraction, the single factor and uniform experimental design and the active ingredients of saffron Optimum extraction process, designed to provide resources for the development of the scientific basis of Xinjiang safflower. An instrument and reagent 1.1 Instrument HP 1100 liquid chromatograph (USA), Shimadzu UV2201 UV spectrophotometer (Japan), high-speed centrifuge (Shanghai Anting), YKH§º liquid fast mixer (Jiangxi), SYZ550 quartz sub-boiling water distiller (Jintan), the United States Thermo Zirchrom C18 column (5 ¦Ìm, 250 mm ¡Á 4.6 mm) (Tianjin Chen Hang Technology Co., Ltd.), a single disk optical analytical balance (Beijing Optical Instrument Factory, d = 0.01 mg) , electric thermostat set (Shanghai Medical Equipment Factory.) 1.2 HSYA reagent A (providing medical department of Peking University, content 98.05%), methanol HPLC grade phosphoric acid were of analytical grade, safflower (Tacheng), ultra-pure water (Shihezi University Hospital Affiliated to self). 2 Methods and Results 2.1 HSYA A determination of chromatographic conditions: mobile phase: methanol -0.5% phosphoric acid (42:58); detection wavelength 400 nm; column temperature 25 ¡æ; flow rate of 1.0 ml / min; injection 5.0 ¦Ìl. External standard method: the number of theoretical plates shall not be less than 6500, resolution greater than 1.5; standard curve (Y = 0.000 375Ai +0.000 526, r = 0.999 8, n = 3), linear relations in the 10 ~ 160 ¦Ìg / ml concentration range can be detected, every 1.0 hours in the day with a known concentration of high, medium and low HSYA standard sample Sample (n = 9) measured the precision (RSD = 1.1%), in each Determination of the test, the sample every 2 hours for Reference Standard HSYA and stability of the test determination (RSD = 1.1%) and reproducibility (RSD = 1.2%), application of high, medium and low concentration of 3 the test for recovery rate experiment, recovery was 101.2%, indicating that the method stable and feasible. Record HSYA chromatograms (Figure 1A). 2.2 Preparation and determination of the test weighed amount of safflower medicine, according to the design extraction process were precise amount of extract 0.5 ml, with a 0.45 ¦Ìm microporous membrane filtration, the filtrate obtained sample determined. Record the test of HSYA chromatograms (Figure 1B). Figure 1HPLC chromatography (HSYA) slightly 2.3 Extraction Process 2.3.1 Single-factor experiment 18.104.22.168 Different indicators of water addition on the extraction times fixed 1, extraction time 1 h, do not soak, each with herbs 6,8,10,15,20 times for reflux extraction of water, HSYA determination results were 0.88%, 1.04%, 1.57%, 1.67%, 1.40%. 22.214.171.124 Indicators of different extraction time on extraction times of 1 fixed, extraction of water for the herbs to 15 times (w / v), do not soak, respectively 20,40,60,80,100 min to reflux extraction, HSYA determination results were 1.32%, 1.49%, 1.45%, 1.44%, 1.25%. 126.96.36.199 Different extraction times on the index of a fixed amount of water is 15 times, the extraction time was 40 min, not soaking, respectively, were extracted by 1,2,3,4 times. HSYA determination results were 1.50%, 1.68%, 1.69%, 1.66%. 188.8.131.52 immersion time on the index of a fixed amount of water is 15 times, the extraction time was 40 min, extraction times of 1 second, respectively, after soaking 0.5,1,2,4,6,8,11 h extraction. HSYA determination results were 1.42%, 1.43%, 1.42%, 1.36%, 1.42%, 1.32%, 1.31%. 2.3.2 Extraction of uniform design 184.108.40.206 Extraction of uniform experimental design can be seen from the single factor experiments solvent volume and extraction time have a greater impact on the index, and extraction times with little effect of immersion time, it will be fixed at 2 times the number of extraction, not immersion. The solvent volume, extraction time is investigated factors, using uniform experiment design selection process. Factor levels in Table 1, experiments, and results in Table 2. Table 1 Experimental design factors uniform level of form factors [omitted] Table 2U5 (52) uniform experiment design table NO. [Omitted] 220.127.116.11 the results of data processing using uniform experiment, Shenyang Pharmaceutical University, uniform design procedures for regression, have equation: Y =- 0.576 0 +0.365 9X1-0.014 2X21 inspection of the equation, residual standard deviation S = 0.009 5, multiple correlation coefficient R2 = 0.994 3, the regression equation test of significance of F = 87.239 6, equation significantly (P <0.05). Optimum conditions: X1 (plus water) = 12.9 times, X2 (extraction time) = 20 min, the theoretical prediction with HSYA 1.78% safflower, prediction interval: 1.62% ~ 1.96%. After a uniform design of experiments and data analysis: determine the process of this study were: the amount of added solvent 13 times the amount of medicine, 100 ¡æ back to the water extraction for 2 times, each time 20 min. 18.104.22.168 verification experiment to take medicine three copies, respectively, determined by this study to extract and sample handling process, using established RPHPLC method for the determination HSYA content of the samples were 1.69% (RSD = 1.18%, n = 3), 1.67% (RSD = 1.25%, n = 3), 1.68% (RSD = 1.58%, n = 3). 3 samples HSYA average content of 1.68% (RSD = 0.60%, n = 3). 3 Discussion 3.1 In the light of the test chemicals and chemical stability in aqueous solution HSYA the visit, the refrigerator -3 ¡æ save 15 d by the ultraviolet and the reproducibility of qualitative and quantitative HPLC chromatogram of the determination of the RSD <2.0%, but room temperature, light, then 48 h chromatographic impurity peak appears, and its UV absorbance and chromatographic peak area of significant variation, RSD> 8.0%. HSYA structure because contains 4 hydroxyl groups, so that stability is affected by temperature, light and other factors larger. 3.2 The research based on the strong yellow water-soluble, water extraction by selecting the first solvent to the active ingredient in HSYA safflower as index, on the extraction process was the experimental study. First screened by single factor indicator of greater impact factor - HSYA (%), and then the level of each factor based on a uniform experimental design, and process optimization. The results show that: Extraction of active ingredients of saffron in the extraction of content can HSYA 1.6% to 1.7%, the method is simple, high yield, time-saving, stable, viable, worthy of promotion, with the development of resources in Xinjiang safflower important application value. References:  National Pharmacopoeia Committee. The Chinese Pharmacopoeia [S]. 2000 edition. A. Beijing: Chemical Industry Press, 2000. 119.  Song Liren, Hong Xun, Ding Xu, et al. Dictionary of Modern Medicine [M]. Beijing: People Health Press, 2001.924928.  Li Guangsheng. On the comprehensive utilization of red flowers [J]. Shanxi Pharmaceutical Industry, 1992, 11 (4): 40.  Takahashi Y, Miyassaka N, Tasaka SH, et al. Constitution of two coloring matters in the flower petals of carthamus tinctorius L [J]. Tetrahedron Lett, 1982,23 (49): 5.
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