[Objectives] To optimize the ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves,and provide raw materials and technical support for development and use of mangiferin relate...[Objectives] To optimize the ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves,and provide raw materials and technical support for development and use of mangiferin related products. [Methods]Five steps( material crushing→ ethyl acetate impurity removing → concentrated extract washing → extracting with methanol → crystallization and precipitation) were used.The single factor experiment and L9( 34) orthogonal experiment was carried out to optimize the process parameters including extraction time,ultrasonic power,extraction times,and extraction temperature.[Results] The optimum process of ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves was as follows: the mango leaves were crushed and sieved; 3 m L/g of ethyl acetate was added,sealed and soaked for 4 h,ultrasonically shaken for 20 min( 50℃,350 W),filtered at room temperature,filtered with 100 mesh sieve,and extracted three times; added 100% methanol to the residue at 3 m L/g,extract by ultrasonic vibration for 20 min( 350 W,55℃)for four times,filtered with 100 mesh sieve when it was still hot; mixed the extract of each time,condensed by vacuum decompression to get the extract; added 100% methanol at 4 m L/g,mixed and washed for 5 min at room temperature,placed for 10 min,filtered with 100 mesh sieve,washed 3 times repeatedly,and dried the filter residue at 60℃ to obtain the crude mangiferin; added 100% methanol at 4 m L/g,mixed and washed at 50℃ for 5 min,placed at 6℃ for 8 h,dried the filter residue at 60℃,and repeatedly crystallized two times. According to the above process,crude and pure mangiferin products could be obtained,the purity of mangiferin of the crude product was higher than 64. 00%,the total recovery rate was 83. 90%,and the purity of mangiferin of the pure product was higher than 98. 00%,and the total recovery rate was about 66. 40%. [Conclusions] The optimized ethyl acetate impurity removal method is easy in operation,low in cost,and high in efficiency for extracting and isolating mangiferin,and can be applied for actual production of mangiferin.展开更多
[Objectives]To explore the effects of ethyl acetate extract of Phyllanthus reticulatus leaves on autophagy-related proteins Beclin-1,ATG5 and LC3 by immunohistochemistry,and to preliminarily explore their effects on a...[Objectives]To explore the effects of ethyl acetate extract of Phyllanthus reticulatus leaves on autophagy-related proteins Beclin-1,ATG5 and LC3 by immunohistochemistry,and to preliminarily explore their effects on autophagy.[Methods]BEL-7404 Hepatocellular Carcinoma(HCC)nude mice model was established,and blank group(same volume of pure water),positive control group(20 mg/kg fluorouracil),high dose drug group(600 mg/kg),and medium dose drug group(300 mg/kg),and low dose drug group(150 mg/kg)were set up.After 2 weeks of intragastric administration,the nude mice were sacrificed,and the tumor tissues were taken out,processed by immunohistochemistry,and then made into paraffin sections.Photos were taken under an optical microscope(10×40),and evaluation and analysis were performed with the aid of the Image-Pro Plus 6.0 image analysis software.Differences were calculated using SPSS 20.0 software.The effects of drugs on autophagy-related proteins LC3,Beclin-1 and ATG5 were observed.[Results]Compared with the blank group,the medium and high dose groups of ethyl acetate extract of P.reticulatus leaves had the effect of promoting the increase of autophagy-related proteins LC3,Beclin-1 and ATG5(P<0.05).However,there was no significant difference between the low dose group of ethyl acetate extract of P.reticulatus leaves and the blank group(P>0.05).[Conclusions]The ethyl acetate extract of P.reticulatus leaves has a promoting effect on autophagy-related proteins LC3,Beclin-1,and ATG5.展开更多
目的:建立番石榴叶提取物中金丝桃苷、异槲皮苷、瑞诺苷、槲皮素-3-O-β-D-吡喃阿拉伯糖苷、槲皮素-3-O-α-L-呋喃阿拉伯糖苷的多成分的含量测定方法。方法:色谱柱:S y n c r o n i s C18色谱柱(4.6mm×250mm,5μm),以乙腈-0.2%磷...目的:建立番石榴叶提取物中金丝桃苷、异槲皮苷、瑞诺苷、槲皮素-3-O-β-D-吡喃阿拉伯糖苷、槲皮素-3-O-α-L-呋喃阿拉伯糖苷的多成分的含量测定方法。方法:色谱柱:S y n c r o n i s C18色谱柱(4.6mm×250mm,5μm),以乙腈-0.2%磷酸水梯度洗脱,流速1.0mL/min,检测波长254nm,柱温40℃。结果:金丝桃苷、异槲皮苷、瑞诺苷、槲皮素-3-O-β-D-吡喃阿拉伯糖苷、槲皮素-3-O-α-L-呋喃阿拉伯糖苷的线性范围为0.0318-0.2544μg(r=0.9998)、0.0575-0.4600μg(r=0.9998)、0.1860-1.4880μg(r=0.9998)、0.1482-1.1856μg(r=0.9998)、0.2012-1.6816μg(r=0.9998),平均加样回收率分别为100.7(RSD=2.6%),100.5(RSD=0.8%),103.4(RSD=2.6%),98.1(RSD=1.7%),100.7(RSD=1.2%),精密度、重复性、稳定性均良好。结论:所建立的方法准确可靠,重复性好,可用于番石榴叶提取物的含量测定。展开更多
基金Supported by Key Technological Innovation Project of Sichuan Province,China(2016XM120)
文摘[Objectives] To optimize the ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves,and provide raw materials and technical support for development and use of mangiferin related products. [Methods]Five steps( material crushing→ ethyl acetate impurity removing → concentrated extract washing → extracting with methanol → crystallization and precipitation) were used.The single factor experiment and L9( 34) orthogonal experiment was carried out to optimize the process parameters including extraction time,ultrasonic power,extraction times,and extraction temperature.[Results] The optimum process of ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves was as follows: the mango leaves were crushed and sieved; 3 m L/g of ethyl acetate was added,sealed and soaked for 4 h,ultrasonically shaken for 20 min( 50℃,350 W),filtered at room temperature,filtered with 100 mesh sieve,and extracted three times; added 100% methanol to the residue at 3 m L/g,extract by ultrasonic vibration for 20 min( 350 W,55℃)for four times,filtered with 100 mesh sieve when it was still hot; mixed the extract of each time,condensed by vacuum decompression to get the extract; added 100% methanol at 4 m L/g,mixed and washed for 5 min at room temperature,placed for 10 min,filtered with 100 mesh sieve,washed 3 times repeatedly,and dried the filter residue at 60℃ to obtain the crude mangiferin; added 100% methanol at 4 m L/g,mixed and washed at 50℃ for 5 min,placed at 6℃ for 8 h,dried the filter residue at 60℃,and repeatedly crystallized two times. According to the above process,crude and pure mangiferin products could be obtained,the purity of mangiferin of the crude product was higher than 64. 00%,the total recovery rate was 83. 90%,and the purity of mangiferin of the pure product was higher than 98. 00%,and the total recovery rate was about 66. 40%. [Conclusions] The optimized ethyl acetate impurity removal method is easy in operation,low in cost,and high in efficiency for extracting and isolating mangiferin,and can be applied for actual production of mangiferin.
基金Supported by the 2018 Basic Ability Improvement Project of Young and Middle-aged Teachers in Guangxi Universities (2018KY0300xp018034)+2 种基金the Education Department of Guangxi Zhuang Autonomous Region and Guangxi University of Traditional Chinese Medicine (2019XK089)Guangxi Science and Technology Base and Talent Project (GuiKe AD20238058)the Key Laboratory of TCM Pharmacology of Guangxi University of Traditional Chinese Medicine
文摘[Objectives]To explore the effects of ethyl acetate extract of Phyllanthus reticulatus leaves on autophagy-related proteins Beclin-1,ATG5 and LC3 by immunohistochemistry,and to preliminarily explore their effects on autophagy.[Methods]BEL-7404 Hepatocellular Carcinoma(HCC)nude mice model was established,and blank group(same volume of pure water),positive control group(20 mg/kg fluorouracil),high dose drug group(600 mg/kg),and medium dose drug group(300 mg/kg),and low dose drug group(150 mg/kg)were set up.After 2 weeks of intragastric administration,the nude mice were sacrificed,and the tumor tissues were taken out,processed by immunohistochemistry,and then made into paraffin sections.Photos were taken under an optical microscope(10×40),and evaluation and analysis were performed with the aid of the Image-Pro Plus 6.0 image analysis software.Differences were calculated using SPSS 20.0 software.The effects of drugs on autophagy-related proteins LC3,Beclin-1 and ATG5 were observed.[Results]Compared with the blank group,the medium and high dose groups of ethyl acetate extract of P.reticulatus leaves had the effect of promoting the increase of autophagy-related proteins LC3,Beclin-1 and ATG5(P<0.05).However,there was no significant difference between the low dose group of ethyl acetate extract of P.reticulatus leaves and the blank group(P>0.05).[Conclusions]The ethyl acetate extract of P.reticulatus leaves has a promoting effect on autophagy-related proteins LC3,Beclin-1,and ATG5.
文摘目的:建立番石榴叶提取物中金丝桃苷、异槲皮苷、瑞诺苷、槲皮素-3-O-β-D-吡喃阿拉伯糖苷、槲皮素-3-O-α-L-呋喃阿拉伯糖苷的多成分的含量测定方法。方法:色谱柱:S y n c r o n i s C18色谱柱(4.6mm×250mm,5μm),以乙腈-0.2%磷酸水梯度洗脱,流速1.0mL/min,检测波长254nm,柱温40℃。结果:金丝桃苷、异槲皮苷、瑞诺苷、槲皮素-3-O-β-D-吡喃阿拉伯糖苷、槲皮素-3-O-α-L-呋喃阿拉伯糖苷的线性范围为0.0318-0.2544μg(r=0.9998)、0.0575-0.4600μg(r=0.9998)、0.1860-1.4880μg(r=0.9998)、0.1482-1.1856μg(r=0.9998)、0.2012-1.6816μg(r=0.9998),平均加样回收率分别为100.7(RSD=2.6%),100.5(RSD=0.8%),103.4(RSD=2.6%),98.1(RSD=1.7%),100.7(RSD=1.2%),精密度、重复性、稳定性均良好。结论:所建立的方法准确可靠,重复性好,可用于番石榴叶提取物的含量测定。
