超临界CO_2流体萃取黄姜中重金属的研究
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摘要
黄姜是我国特有经济作物，其有效成分甾体皂素是合成甾体激素药物的基础原料，拥有“药用黄金”之美誉。为降低黄姜中重金属含量并保护其有效成分，本论文进行了超临界CO_2流体络合萃取法净化黄姜中Cu、Cd、Pb的工艺研究，并探讨了黄姜中重金属净化后对其有效成分的影响。主要研究内容及结果如下：
1、黄姜中重金属Cu、Cd、Pb的含量测定
(1)通过比较微波消解和湿法消解两种样品前处理方式发现，前者消解效果更好，速度快，试剂用量少，测定结果更准确。
(2)通过优化仪器操作参数，建立了微波消解—火焰原子吸收和微波消解—石墨炉原子吸收测定黄姜中Cu、Cd、Pb的方法。在优化后的条件下测得黄姜中Cu含量为11.51μg／g，Cd含量为0.28μg╱g，Pb含量为3.60μg╱g。
2、超临界CO_2流体络合萃取黄姜中Cu、Cd、Pb的方法
(1)采用单因素试验法对络合剂、络合方式、萃取压力、温度、时间、改性剂含量等因素的影响进行了考察。试验表明，二乙基二硫代氨基甲酸钠作络合剂效果较好；其络合方式为萃取前加入到样品中密闭放置1h；各条件因素对重金属萃取率的影响在开始阶段均表现为正相关。
(2)综合单因素试验结果，采用L_9(3~4)正交表设计试验，结果分析表明，各因素的影响程度为：萃取压力＞萃取温度＞萃取时间＞改性剂含量。
(3)根据正交试验结果，采用中心组合设计法(三因素三水平响应面分析)对萃取条件进行了优化，建立了重金属净化率的二次回归方程。此模型拟合性好，通过响应面分析及岭嵴分析得到了优化组合条件。结果表明，压力、温度、时间均对净化率有极显著影响。当萃取条件为压力25.54MPa，温度59℃，时间35min时，黄姜中重金属净化率达到极大值。此条件下净化率预测值为71.71％，验证值为75.10％。净化后的黄姜达到了重金属国际限量标准。
3、重金属净化后对黄姜中有效成分的影响
(1)超临界CO_2流体萃取法与有机溶剂萃取法对比表明，前者对黄姜中甾体皂素的提取效果更好，萃取率较高，且安全高效、工艺简单、操作温度低、无溶剂污染。
(2)采用超临界CO_2流体萃取法提取黄姜中甾体皂素，以可见分光光度法测定其含量，得到黄姜原样品及重金属净化后样品中甾体皂素含量分别为2.03％和1.84％，超临界CO_2流体络合萃取黄姜中Cu、Cd、Pb后对其有效成分影响不大。
Dioscorea Zingiberensis C. H. Wright is an important unique economic plant, whoseactive ingredient is called sapogenins and used in the synthesis of the body hormonemedicine as the foundation raw material, which makes it called officinal gold. In order toreduce the content of heavy metals in Dioscorea Zingiberensis C. H. Wright, and protectthe active ingredient, the technology of extracting Cu, Cd, Pb by supercritical CO_2 fluid,and the influence of depuration of heavy metals on the active ingredient were investigated.The main contents and results are listed as follows:
1. Determination of Cu, Cd, Pb in Dioscorea Zingiberensis C. H. Wright
(1) The microwave digestion method was investigated and compared with theorganic solvent extraction method. Experimental results showed that, the former methodhad the advantages of faster speed, less usage amount of digestive dissolvent and higheraccuracy.
(2) Microwave digestion-flame/graphite furnace atomic absorption spectrometrymethods were established for the determination of Cu, Cd, Pb in Dioscorea ZingiberensisC. H. Wright. The content of Cu, Cd, Pb was 3.60μg/g, 0.28μg/g and 11.51μg/grespectively under optimum conditions.
2. Supercritical CO_2 fluid chelating extraction of Cu, Cd, Pb from DioscoreaZingiberensis C. H. Wright
(1) The effects of chelating agent, chelating mode, extraction pressure, temperature,time and content of modifier on extraction yield were investigated. Experimental resultsshowed that, sodium diethyldithiocarbamate was the best used as the chelating agent forthe extraction of heavy metals. It must be mixed into the sample before extraction for 1h.Extraction pressure, temperature, time and content of modifier all had positivecorrelation with extraction yield in intial stage.
(2) Four-factor and three-level orthogonal design was used to optimize the extractionconditions. The influence of various factors was in the order: pressure＞temperature＞time＞content of modifier.
(3) Based on orthogonal test, three levels of the factors were selected to the centralcomposite design experiment. The quadric regression model for the purifying rate wasestablished. The model was significantly fit well and the optimum combination wasobtained by response surface analysis and ridge analysis. Results showed that, pressure, temperature and time were all significant to the purifying rate. When the extractionparameters were composed of 25.54MPa, 59℃and 35min, the purifying rate reachedmaximum, which was verified to 75.10% and predicted to 71.71%. After depuration, thecontent of heavy metals in Dioscorea Zingiberensis C. H. Wright reached the limitedquantity of international standard.
3. Influence of depuration of heavy metals in Dioscorea Zingiberensis C. H.Wright on the active ingredient
(1) Supercritical CO_2 fluid chelating extraction was investigated and compared withthe organic solvent extraction method. Experimental results showed that, the formermethod had the advantages of high recoveries, safety, efficiency, simple crafts, lowoperating temperature and clean.
(2) The ultraviolet spectrophotometry method as determining the content ofsapogenins was used to evaluate the change of the quality in Dioscorea Zingiberensis C.H. Wright before or after supercritical CO_2 fluid extraction. The content of sapogeninswas 2.03% and 1.84% respectively in original and depurative sample, which showed thatdepuration of Cu, Cd, Pb in Dioscorea Zingiberensis C. H. Wright by supercritical CO_2fluid extraction had little influence on the active ingredient.
1、黄姜中重金属Cu、Cd、Pb的含量测定
(1)通过比较微波消解和湿法消解两种样品前处理方式发现，前者消解效果更好，速度快，试剂用量少，测定结果更准确。
(2)通过优化仪器操作参数，建立了微波消解—火焰原子吸收和微波消解—石墨炉原子吸收测定黄姜中Cu、Cd、Pb的方法。在优化后的条件下测得黄姜中Cu含量为11.51μg／g，Cd含量为0.28μg╱g，Pb含量为3.60μg╱g。
2、超临界CO_2流体络合萃取黄姜中Cu、Cd、Pb的方法
(1)采用单因素试验法对络合剂、络合方式、萃取压力、温度、时间、改性剂含量等因素的影响进行了考察。试验表明，二乙基二硫代氨基甲酸钠作络合剂效果较好；其络合方式为萃取前加入到样品中密闭放置1h；各条件因素对重金属萃取率的影响在开始阶段均表现为正相关。
(2)综合单因素试验结果，采用L_9(3~4)正交表设计试验，结果分析表明，各因素的影响程度为：萃取压力＞萃取温度＞萃取时间＞改性剂含量。
(3)根据正交试验结果，采用中心组合设计法(三因素三水平响应面分析)对萃取条件进行了优化，建立了重金属净化率的二次回归方程。此模型拟合性好，通过响应面分析及岭嵴分析得到了优化组合条件。结果表明，压力、温度、时间均对净化率有极显著影响。当萃取条件为压力25.54MPa，温度59℃，时间35min时，黄姜中重金属净化率达到极大值。此条件下净化率预测值为71.71％，验证值为75.10％。净化后的黄姜达到了重金属国际限量标准。
3、重金属净化后对黄姜中有效成分的影响
(1)超临界CO_2流体萃取法与有机溶剂萃取法对比表明，前者对黄姜中甾体皂素的提取效果更好，萃取率较高，且安全高效、工艺简单、操作温度低、无溶剂污染。
(2)采用超临界CO_2流体萃取法提取黄姜中甾体皂素，以可见分光光度法测定其含量，得到黄姜原样品及重金属净化后样品中甾体皂素含量分别为2.03％和1.84％，超临界CO_2流体络合萃取黄姜中Cu、Cd、Pb后对其有效成分影响不大。
Dioscorea Zingiberensis C. H. Wright is an important unique economic plant, whoseactive ingredient is called sapogenins and used in the synthesis of the body hormonemedicine as the foundation raw material, which makes it called officinal gold. In order toreduce the content of heavy metals in Dioscorea Zingiberensis C. H. Wright, and protectthe active ingredient, the technology of extracting Cu, Cd, Pb by supercritical CO_2 fluid,and the influence of depuration of heavy metals on the active ingredient were investigated.The main contents and results are listed as follows:
1. Determination of Cu, Cd, Pb in Dioscorea Zingiberensis C. H. Wright
(1) The microwave digestion method was investigated and compared with theorganic solvent extraction method. Experimental results showed that, the former methodhad the advantages of faster speed, less usage amount of digestive dissolvent and higheraccuracy.
(2) Microwave digestion-flame/graphite furnace atomic absorption spectrometrymethods were established for the determination of Cu, Cd, Pb in Dioscorea ZingiberensisC. H. Wright. The content of Cu, Cd, Pb was 3.60μg/g, 0.28μg/g and 11.51μg/grespectively under optimum conditions.
2. Supercritical CO_2 fluid chelating extraction of Cu, Cd, Pb from DioscoreaZingiberensis C. H. Wright
(1) The effects of chelating agent, chelating mode, extraction pressure, temperature,time and content of modifier on extraction yield were investigated. Experimental resultsshowed that, sodium diethyldithiocarbamate was the best used as the chelating agent forthe extraction of heavy metals. It must be mixed into the sample before extraction for 1h.Extraction pressure, temperature, time and content of modifier all had positivecorrelation with extraction yield in intial stage.
(2) Four-factor and three-level orthogonal design was used to optimize the extractionconditions. The influence of various factors was in the order: pressure＞temperature＞time＞content of modifier.
(3) Based on orthogonal test, three levels of the factors were selected to the centralcomposite design experiment. The quadric regression model for the purifying rate wasestablished. The model was significantly fit well and the optimum combination wasobtained by response surface analysis and ridge analysis. Results showed that, pressure, temperature and time were all significant to the purifying rate. When the extractionparameters were composed of 25.54MPa, 59℃and 35min, the purifying rate reachedmaximum, which was verified to 75.10% and predicted to 71.71%. After depuration, thecontent of heavy metals in Dioscorea Zingiberensis C. H. Wright reached the limitedquantity of international standard.
3. Influence of depuration of heavy metals in Dioscorea Zingiberensis C. H.Wright on the active ingredient
(1) Supercritical CO_2 fluid chelating extraction was investigated and compared withthe organic solvent extraction method. Experimental results showed that, the formermethod had the advantages of high recoveries, safety, efficiency, simple crafts, lowoperating temperature and clean.
(2) The ultraviolet spectrophotometry method as determining the content ofsapogenins was used to evaluate the change of the quality in Dioscorea Zingiberensis C.H. Wright before or after supercritical CO_2 fluid extraction. The content of sapogeninswas 2.03% and 1.84% respectively in original and depurative sample, which showed thatdepuration of Cu, Cd, Pb in Dioscorea Zingiberensis C. H. Wright by supercritical CO_2fluid extraction had little influence on the active ingredient.
引文
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77. Smart N G, Carleson T E, Elshani S, Wang S, Wai C M. Extraction of toxic metals using supercritical fluid carbon dioxide containing organophosphorus reagents. Ind Eng Chem Res, 1997,36(5): 1819-1826
78. Tai C Y, You G S, Chen S L. Kinetics study on supercritical fluid extraction of zinc(II) ion from aqueous solutions. Supercrit Fluids, 2000, 18(3): 201-212
79. Takeshita Y, Sato Y, Nishi S. Study of extraction of metals from CCA-treated wood with supercritical CO2 containing acetylacetone: extraction of Cu by continuous addition of acetylacetone. Ind Eng Chem Res, 2000, 39(12): 4496-4499
80. Takeshita Y, Sato Y, Nishi S. A study of feasibility of extracting metals from CCA-treated woods by using supercritical carbondioxide. Waste Manage Res, 2000, 11(2): 94-100
81. Toews K L, Shroll R M, Wai C M, Smart N G. pH-defining equilibrium between water and supercritical CO2. Influence on SFE of organics and metal chelates. Anal Chem, 1995, 67 (22): 4040-4043
82. Tomioka O, Enokida Y, Yamamoto I. Solvent extraction of lanthanides from their xides with TBP in supercritical carbon dioxide. Nucl Sci Technol, 1998, 35(7): 515-516
83. Tomioka O, Enokida Y, Yamamoto I, Takahash T. Cleaning of materials contaminated with metal oxides through supercritical fluid extraction with CO2 containing TBP. Progress in Nuclear Energy, 2000, 37(1-4): 417-422
84. Trofimov T I, Samsonov M D, Kulyako Y M, Myasoedov B F. Comptes Rendus Chimie, 2004, 7(12): 1209-1213
85. Wai C M, Lin Y, Brauer R, Wang S, Beckert W F. Supercritical fluid extraction of organic and inorganic mercury from solid materials. Talanta, 1993, 40(9): 1325-1330
86. Wai C M, Wang S, Liu Y, Lopez-Avila V, Beckert W F. Evaluation of dithiocarbamates and β-diketones as chelating agents in supercritical fluid extraction of Cd, Pb and Hg from solid samples. Tanlanta,1996, 43(12): 2083-2091
87. Wai C M, Wang S.Supercritical fluid extraction: metal as complexes. J Chromatogr A, 1997, 785(1-2): 369-383
88. Wang J, Marshall W D. Recovery of metals from aqueous media by extraction with supercritical carbon dioxide. Anal Chem, 1994,66: 1658-1663
89. Wang S, Elshani S, Wai C M. Selective extraction of mercury with ionizable crown ethers in supercritical carbon dioxide. Anal Chem, 1995, 67(5): 919-923
90. Wenclawiak B W, Hees T, Zoller C E, Kabus H P. Rhodium and palladium P-diketonate determination with on-line supercritical fluid extraction-high performance liquid chromatography via solid phase extraction. Anal Chem, 1997, 358(4):471-474
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76. Shu X, Gao Z, Yang X. Supercritical fluid extraction of sapogenins from tubers of Smilax china. Fitoterapia, 2004, 75: 656-661
77. Smart N G, Carleson T E, Elshani S, Wang S, Wai C M. Extraction of toxic metals using supercritical fluid carbon dioxide containing organophosphorus reagents. Ind Eng Chem Res, 1997,36(5): 1819-1826
78. Tai C Y, You G S, Chen S L. Kinetics study on supercritical fluid extraction of zinc(II) ion from aqueous solutions. Supercrit Fluids, 2000, 18(3): 201-212
79. Takeshita Y, Sato Y, Nishi S. Study of extraction of metals from CCA-treated wood with supercritical CO2 containing acetylacetone: extraction of Cu by continuous addition of acetylacetone. Ind Eng Chem Res, 2000, 39(12): 4496-4499
80. Takeshita Y, Sato Y, Nishi S. A study of feasibility of extracting metals from CCA-treated woods by using supercritical carbondioxide. Waste Manage Res, 2000, 11(2): 94-100
81. Toews K L, Shroll R M, Wai C M, Smart N G. pH-defining equilibrium between water and supercritical CO2. Influence on SFE of organics and metal chelates. Anal Chem, 1995, 67 (22): 4040-4043
82. Tomioka O, Enokida Y, Yamamoto I. Solvent extraction of lanthanides from their xides with TBP in supercritical carbon dioxide. Nucl Sci Technol, 1998, 35(7): 515-516
83. Tomioka O, Enokida Y, Yamamoto I, Takahash T. Cleaning of materials contaminated with metal oxides through supercritical fluid extraction with CO2 containing TBP. Progress in Nuclear Energy, 2000, 37(1-4): 417-422
84. Trofimov T I, Samsonov M D, Kulyako Y M, Myasoedov B F. Comptes Rendus Chimie, 2004, 7(12): 1209-1213
85. Wai C M, Lin Y, Brauer R, Wang S, Beckert W F. Supercritical fluid extraction of organic and inorganic mercury from solid materials. Talanta, 1993, 40(9): 1325-1330
86. Wai C M, Wang S, Liu Y, Lopez-Avila V, Beckert W F. Evaluation of dithiocarbamates and β-diketones as chelating agents in supercritical fluid extraction of Cd, Pb and Hg from solid samples. Tanlanta,1996, 43(12): 2083-2091
87. Wai C M, Wang S.Supercritical fluid extraction: metal as complexes. J Chromatogr A, 1997, 785(1-2): 369-383
88. Wang J, Marshall W D. Recovery of metals from aqueous media by extraction with supercritical carbon dioxide. Anal Chem, 1994,66: 1658-1663
89. Wang S, Elshani S, Wai C M. Selective extraction of mercury with ionizable crown ethers in supercritical carbon dioxide. Anal Chem, 1995, 67(5): 919-923
90. Wenclawiak B W, Hees T, Zoller C E, Kabus H P. Rhodium and palladium P-diketonate determination with on-line supercritical fluid extraction-high performance liquid chromatography via solid phase extraction. Anal Chem, 1997, 358(4):471-474