不同浸提方式下HPLC测定苹果渣中cAMP的研究
|
摘要
以苹果渣为原料,用蒸馏水浸提法和20%乙醇溶液浸提法分别提取苹果中cAMP,并利用HPLC方法进行苹果渣中cAMP的定性、定量分析。HPLC条件:流动相为甲醇-双蒸水,检测波长为259nm,流速1.0mL/min。采用保留时间法定性,相关系数R2为0.9997;精密度实验中RSD为1.01%;重复性实验中RSD为1.67%;回收率实验中cAMP平均回收率为97.64%,RSD为1.92%。水浸提法测定的苹果渣中cAMP含量为(225.8401±4.9221)μg/g,乙醇溶液浸提法测定苹果渣中cAMP含量为(132.5300±5.4507)μg/g。
The qualitative and quantitative analysis of cAMP using distilled water extraction and 20% ethanol solution extraction to extract cAMP from apple pomace were studied in this paper.The HPLC method was adopted to detect cAMP content in the extract.The methanol and double distilled water was the mobile phase of HPLC.The wavelength was 259nm and the flow rate was 1.0mL/min.In the qualitative test using retention time method,the correlation coefficients(R2) was 0.9997.The relative standard deviations(RSD) were 1.01% and 1.67% in precision experiment and reproducibility experiment,respectively.In the experiment of recovery rate,the average recovery rate of cAMP was 97.64% and RSD was 1.92%.The result of quantitative test showed the cAMP content was(225.8401 ±4.9221)μg/g with distilled water extraction in apple pomace,meanwhile it was(132.5300±5.4507)μg/g with ethanol solution extraction.
The qualitative and quantitative analysis of cAMP using distilled water extraction and 20% ethanol solution extraction to extract cAMP from apple pomace were studied in this paper.The HPLC method was adopted to detect cAMP content in the extract.The methanol and double distilled water was the mobile phase of HPLC.The wavelength was 259nm and the flow rate was 1.0mL/min.In the qualitative test using retention time method,the correlation coefficients(R2) was 0.9997.The relative standard deviations(RSD) were 1.01% and 1.67% in precision experiment and reproducibility experiment,respectively.In the experiment of recovery rate,the average recovery rate of cAMP was 97.64% and RSD was 1.92%.The result of quantitative test showed the cAMP content was(225.8401 ±4.9221)μg/g with distilled water extraction in apple pomace,meanwhile it was(132.5300±5.4507)μg/g with ethanol solution extraction.
引文
[1]唐雪,马海田,邹思湘.脱氢表雄酮(DHEA)对鸡胚原代肝细胞cAMP/PKA信号通路和cAMP反应元件结合蛋白(CREB)的影响与调节[J].农业生物技术学报,2012,20(6):649-656.
[2]Miao J,Fang S,Bae Y,et al.Functional inhibitory cross-talk between constitutive androstane receptor and hepatic nuclear factor-4 in hepatic lipid/glucose metabolism is mediated by competition for binding to the DR1 motif and to the common coactivators,GRIP-1 and PGC-1alpha[J].Journal of Biological Chemistry,2006,281(21):14537-14546.
[3]杨夏,彭生,刘功俭.cAMP/PKA/CREB信号通路及相关调控蛋白PDE-4和ERK对学习记忆的影响[J].医学综述,2011,17(15):2241-2243.
[4]Amrhein N.Evidence against the occurrence of adenosine3’,5’-cyclic monophosphate in higher plants[J].Plarrta,1974,118(3):241-258.
[5]Amrhein N.The current status of cyclic AMP in higher plants[J].Anrnal Review of Plant Physiology,1977(28):123-132.
[6]Hanabusa K,Cyong J,Takahashi M.High-level of cyclic AMP in the jujube plum[J].Planta Medica,1981,42:380-384.
[7]Jyong-Chyul Cyong,Kiyomichi Hanabusa.Cyclic adenosine monophosphate in fruits of zizyphus jujuba[J].Pytochemistry,1980(19):2747-2748.
[8]Jyong-Chyul Cyong,Makoto Takahashi.Identification of guanosine 3’,5’-monophosphate in the fruit of zizyphus jujuba[J].Phytochernistry,1982(21):1871-1874.
[9]张明娟,李薇,庞晓明.枣果中环磷酸腺苷cAMP的提取工艺及含量测定[J].食品与发酵工业,2012,38(5):228-231.
[10]刘孟军,王永蕙.枣和酸枣等14种园艺植物cAMP含量的研究[J].河北农业大学学报,1991,14(4):20-23.
[11]Kaushal Nk,Joshi VK,Sharma RC.Effect of stages of apple pomace collection and treatment on physic-chemical and sensory qualities of pomace leather[J].Journal of Food Science and Technology,2002,39(4):388-393.
[12]Francielo V,Patrcia MA,Fernanda S,et al.Apple pomance:a versatile substrate for biotechnology applications[J].Critical Reviews in Biotechnology,2008,28(1):1-12.
[13]于滨,吴茂玉,朱凤涛,等.苹果渣综合利用研究进展[J].中国果蔬,2012(12):31-34.
[14]张倩,樊军,罗云伟.HPLC测定陕北大枣和新疆大枣中环磷酸腺苷含量的研究[J].药物与分析杂志,2008,28(6):895-897.
[15]史景江,熙中.色谱分析法[M].重庆:重庆大学出版社,1995:182-242.
[16]王立霞,陈锦屏,张娜,等.和田玉枣中cAMP提取工艺的研究[J].食品工业科技,2009,30(10):234-236.
[17]蒲云峰,万英,侯旭杰.HPLC法测定不同品种红枣中cAMP含量[J].食品研究与开发,2011,32(7):109-112.
[18]张爱华,杨宝华.cAMP的构象及其异构化反应的理论研究[J].信阳师范学院学报:自然科学版,2012,25(4):461-464.
[19]王立霞,陈锦屏,张娜,等.和田玉枣中环磷酸腺苷(cAMP)的分离纯化工艺研究[J].食品科学,2008,29(12):250-254.
[20]夏全明,周洋,赵黎明,等.离子交换法提取大枣中环磷酸腺苷(cAMP)的工艺研究[J].生物技术进展,2012,2(4):288-292.
[2]Miao J,Fang S,Bae Y,et al.Functional inhibitory cross-talk between constitutive androstane receptor and hepatic nuclear factor-4 in hepatic lipid/glucose metabolism is mediated by competition for binding to the DR1 motif and to the common coactivators,GRIP-1 and PGC-1alpha[J].Journal of Biological Chemistry,2006,281(21):14537-14546.
[3]杨夏,彭生,刘功俭.cAMP/PKA/CREB信号通路及相关调控蛋白PDE-4和ERK对学习记忆的影响[J].医学综述,2011,17(15):2241-2243.
[4]Amrhein N.Evidence against the occurrence of adenosine3’,5’-cyclic monophosphate in higher plants[J].Plarrta,1974,118(3):241-258.
[5]Amrhein N.The current status of cyclic AMP in higher plants[J].Anrnal Review of Plant Physiology,1977(28):123-132.
[6]Hanabusa K,Cyong J,Takahashi M.High-level of cyclic AMP in the jujube plum[J].Planta Medica,1981,42:380-384.
[7]Jyong-Chyul Cyong,Kiyomichi Hanabusa.Cyclic adenosine monophosphate in fruits of zizyphus jujuba[J].Pytochemistry,1980(19):2747-2748.
[8]Jyong-Chyul Cyong,Makoto Takahashi.Identification of guanosine 3’,5’-monophosphate in the fruit of zizyphus jujuba[J].Phytochernistry,1982(21):1871-1874.
[9]张明娟,李薇,庞晓明.枣果中环磷酸腺苷cAMP的提取工艺及含量测定[J].食品与发酵工业,2012,38(5):228-231.
[10]刘孟军,王永蕙.枣和酸枣等14种园艺植物cAMP含量的研究[J].河北农业大学学报,1991,14(4):20-23.
[11]Kaushal Nk,Joshi VK,Sharma RC.Effect of stages of apple pomace collection and treatment on physic-chemical and sensory qualities of pomace leather[J].Journal of Food Science and Technology,2002,39(4):388-393.
[12]Francielo V,Patrcia MA,Fernanda S,et al.Apple pomance:a versatile substrate for biotechnology applications[J].Critical Reviews in Biotechnology,2008,28(1):1-12.
[13]于滨,吴茂玉,朱凤涛,等.苹果渣综合利用研究进展[J].中国果蔬,2012(12):31-34.
[14]张倩,樊军,罗云伟.HPLC测定陕北大枣和新疆大枣中环磷酸腺苷含量的研究[J].药物与分析杂志,2008,28(6):895-897.
[15]史景江,熙中.色谱分析法[M].重庆:重庆大学出版社,1995:182-242.
[16]王立霞,陈锦屏,张娜,等.和田玉枣中cAMP提取工艺的研究[J].食品工业科技,2009,30(10):234-236.
[17]蒲云峰,万英,侯旭杰.HPLC法测定不同品种红枣中cAMP含量[J].食品研究与开发,2011,32(7):109-112.
[18]张爱华,杨宝华.cAMP的构象及其异构化反应的理论研究[J].信阳师范学院学报:自然科学版,2012,25(4):461-464.
[19]王立霞,陈锦屏,张娜,等.和田玉枣中环磷酸腺苷(cAMP)的分离纯化工艺研究[J].食品科学,2008,29(12):250-254.
[20]夏全明,周洋,赵黎明,等.离子交换法提取大枣中环磷酸腺苷(cAMP)的工艺研究[J].生物技术进展,2012,2(4):288-292.