建立高效液相色谱—双波长同时检测咖啡豆提取物中绿原酸和咖啡因含量的方法。咖啡因和绿原酸在0.2～1.0 mg·mL-1范围内，峰面积与浓度呈现良好的线性关系。咖啡因：Y＝44077X＋227.78，R＝0.9997，RSD＝0.78%；绿原酸：Y＝34896X＋289.96，R＝0.9995, RSD＝0.93%。以绿原酸含量为指标，在单因素试验基础上建立正交试验，确定从绿咖啡豆中提取绿原酸的最佳条件为乙醇浓度20%，液料比6∶1，提取时间60 min，提取温度70 ℃。以咖啡因和绿原酸含量为指标，通过萃取试验，确定最佳脱咖啡因萃取剂为环己烷-三氯甲烷-丙酮混合溶剂，萃取工艺为1.5倍体积萃取剂萃取3次，产物中咖啡因含量由(3.52±0.08)%下降至(0.47±0.11)%。
The method of high performance liquid chromatography with dual wavelength was established for simultaneous determination of caffeine and chlorogenic acid in coffee bean extracts. The result showed a good linear relationship between peak area and concentration in the range of 0.2—1.0 mg·mL-1. The regression equations of caffeine were: Y=44077X+227.78, R=0.9997, RSD=0.78%; the regression equations of chlorogenic acid were: Y=34896X+289.96, R=0.9995, RSD=0.93%. With chlorogenic acid content as indicator, the optimal conditions for extracting chlorogenic acid from green coffee beans were determined as follows: ethanol concentration was 20%, solid-liquid ratio was 6:1, extraction time was 60 min, the extraction temperature was 70 ℃ according to orthogonal test and single factor test. Taking caffeine and chlorogenic acid contents as indicators, the best decaffeination extractant was cyclohexane-trichloromethane-acetone mixture, and the extraction process was extracted three times with 1.5 volumes of cyclohexane-trichloromethane- acetone mixture with the caffeine content decreased from (3.52±0.08)% to (0.47±0.11)%.
[1] Priftis A, Stagos D, Konstantinopoulos K, Tsitsimpikou C, Spandidos D A, Tsatsakis A M, Tzatzarakis M N, Kouretas D. Comparison of antioxidant activity between green and roasted coffee beans using molecular methods[J]. Molecular Medicine Reports, 2015,12(5): 7293—7302.
[2] Kang J, Liu Y, Xie M X, Li S, Jiang M, Wang Y D. Interactions of human serum albumin with chlorogenic acid and ferulic acid[J]. Biochimica et Biophysica Acta, 2004,1674(2): 205—214.
[3] 杨剀舟. 绿原酸抗氧化活性及其自协同抗氧化机理研究[D]. 北京: 中国农业大学博士学位论文, 2016.
[4] 胡仲秋,洪小迪,岳田利. 光皮木瓜绿原酸的提取及抗菌活性测定[J]. 食品科学, 2010,31(24): 8—13.
[5] Ong K W, Hsu A, Tan B K H. Anti-diabetic and anti-lipidemic effects of chlorogenic acid are mediated by ampk activation[J]. Biochemical Pharmacology, 2013,85(9): 1341—1351.
[6] 龙文静. 咖啡豆中多酚类物质的提制技术及抗氧化性能评价[D]. 长沙: 湖南农业大学硕士学位论文, 2011.
[7] 潘晓靓,刘冬琳,孟巍,李婧婷,平洁,汪晖. 咖啡因暴露对小鼠胚胎发育的影响[J]. 武汉大学学报(医学版), 2008,29(2): 165—168.
[8] Smith A. Effects of caffeine on human behavior[J]. Food and Chemical Toxicology, 2002,40(9): 1243—1255.
[9] Shilo L, Sabbah H, Hadari R, Kovatz S, Weinberg U, Dolev S, Dagan Y, Shenkman L. The effects of coffee consumption on sleep and melatonin secretion[J]. Sleep Medicine, 2002,3(3): 271—273.
[10] Lane J D, Pieper C F, Phillips-Bute B G, Bryant J E, Kuhn C M. Caffeine affects cardiovascular and neuroendocrine activation at work and home[J]. Psychosomatic Medicine, 2002,64(4): 595—603.
[11] 张清林. 咖啡和咖啡因的致突变作用[J]. 中国公共卫生, 1995,11(4): 162—164.
[12] Wen W, Shu X O, Jacobs Jr D R, Brown J E. The associations of maternal caffeine consumption and nausea with spontaneous abortion[J]. Epidemiology, 2001,12(1): 38—42.
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