The Content Analysis of Gastrodin and Gastrodigenin Obtained by Different Processing Methods

The Content Analysis of Gastrodin and Gastrodigenin Obtained by Different Processing Methods Abstract The purpose of this article was to investigate the effects of different processing methods on the content of gastrodin and gastrodigenin in Gastrodia elata Blume, so as to obtain the best processing method of G. elata. The contents of gastrodin and gastrodigenin in the samples were determined by HPLC method in turn. The results showed that the content of gastrodin and gastrodigenin were the highest in the sample obtained by extraction with 50% ethanol after boiling. Therefore, according to the two indexes of gastrodin and gastrodigenin, alcohol extraction after boiling was the best processing method, which could provide the reference for production. Introduction Gastrodia elata Blume is the subterranean dried tuber of perennial herbaceous plant in Orchidaceae (1). The gastrodin and gastrodigenin in G. elata are the main active ingredients (2), and they have specific pharmacological activities, which allow their use as medicines (3). Gastrodin has sedative, anticonvulsant, analgesic and hypnotic effects (4). Gastrodigenin has the same analgesic effect as gastrodin (5). At present, the best processing method of G. elata is based on the determination of gastrodin content (6, 7). In this study, fresh G. elata Blume was used as raw material. It was processed by water extraction after boiling, alcohol extraction after boiling, water extraction after steaming, alcohol extraction after steaming, water extraction after baking and alcohol extraction after baking, respectively. The content of gastrodin and gastrodigenin was determined by HPLC. The aim was to observe the dynamic balance of gastrodin and gastrodigenin. At the same time, it would provide a scientific and effective method for the processing of G. elata. Experimental General The samples were analyzed by Hitachi high performance liquid chromatography L-2000. Gastrodia elata (fresh) was collected from Chengkou county of Chongqing city, China. The sampling time was on 28 March 2017. Acetonitrile (MeCN) was of chromatographic-purity grade. Purified water was made at our lab. Gastrodin control, gastrodigenin control and other reagents were of analytical-reagent grade. Chromatographic condition Chromatographic column: Lachrom 891-5050; mobile phase: acetonitrile–0.05% phosphoric acid solution (3:97); detection wavelength: 220 nm; flow rate: 1.0 mL/min; column temperature: 40°C; sample size: 20 μL. The processing method of G. elata The same volume of fresh G. elata was collected, which was evenly divided into three portions. The first portion of fresh G. elata was put into boiling water and cooked thoroughly, then sliced and dried at 60°C in oven for next use. The second portion of fresh G. elata was put into the steamer and steamed thoroughly, then sliced and dried at 60°C in oven for next use. The third portion of fresh G. elata was put into the oven and heated thoroughly at 110°C, then sliced and dried at 60°C in oven for next use. Preparation of sample solution To take and crush a certain amount of the above processed sample in turn, add 50% ethanol or pure water (50 mL) and weigh. To be refluxed for 3 h and cooled. To weigh and supplement lost weight with extraction solvent. To shake and filtrate. The filtrate (20 mL) was concentrated to dryness, and 20% acetonitrile water solution was added. To be dissolved in 50 mL volumetric flask and diluted to the scale. To shake and be filtrated with microporous membrane (0.45 μm). So, the test sample solution was obtained. Results In the samples obtained by different processing methods, the content determination results of gastrodin and gastrodigenin were shown in Table I. Table I. The Content Determination Results of Gastrodin and Gastrodigenin in Samples Obtained by Different Processing Methods Sample source  Test item  Measured content (μg)  Sample size (g)  Measured content (g)/sample size (g)  Water extraction after boiling  Gastrodin  114.3359412 ± 0.0000628  0.0221  0.00517 ± 0.00063    Gastrodigenin  70.37750841 ± 0.00007323    0.00318 ± 0.00057  Alcohol extraction after boiling  Gastrodin  156.1118825 ± 0.0000195  0.023  0.00679 ± 0.00017    Gastrodigenin  258.7135519 ± 0.0000332    0.01125 ± 0.00092  Water extraction after steaming  Gastrodin  93.68121212 ± 0.0000715  0.0208  0.0045 ± 0.00068    Gastrodigenin  52.71591707 ± 0.00000746    0.00253 ± 0.00055  Alcohol extraction after steaming  Gastrodin  142.8957821 ± 0.0000162  0.0213  0.00671 ± 0.00026    Gastrodigenin  132.5645811 ± 0.0000276    0.00622 ± 0.00059  Water extraction after baking  Gastrodin  32.21564738 ± 0.00002738  0.0216  0.00149 ± 0.00085    Gastrodigenin  78.5830962 ± 0.00001835    0.00364 ± 0.00093  Alcohol extraction after baking  Gastrodin  32.10239363 ± 0.00001126  0.0298  0.00108 ± 0.00026    Gastrodigenin  325.148749 ± 0.000062    0.01091 ± 0.00073  Sample source  Test item  Measured content (μg)  Sample size (g)  Measured content (g)/sample size (g)  Water extraction after boiling  Gastrodin  114.3359412 ± 0.0000628  0.0221  0.00517 ± 0.00063    Gastrodigenin  70.37750841 ± 0.00007323    0.00318 ± 0.00057  Alcohol extraction after boiling  Gastrodin  156.1118825 ± 0.0000195  0.023  0.00679 ± 0.00017    Gastrodigenin  258.7135519 ± 0.0000332    0.01125 ± 0.00092  Water extraction after steaming  Gastrodin  93.68121212 ± 0.0000715  0.0208  0.0045 ± 0.00068    Gastrodigenin  52.71591707 ± 0.00000746    0.00253 ± 0.00055  Alcohol extraction after steaming  Gastrodin  142.8957821 ± 0.0000162  0.0213  0.00671 ± 0.00026    Gastrodigenin  132.5645811 ± 0.0000276    0.00622 ± 0.00059  Water extraction after baking  Gastrodin  32.21564738 ± 0.00002738  0.0216  0.00149 ± 0.00085    Gastrodigenin  78.5830962 ± 0.00001835    0.00364 ± 0.00093  Alcohol extraction after baking  Gastrodin  32.10239363 ± 0.00001126  0.0298  0.00108 ± 0.00026    Gastrodigenin  325.148749 ± 0.000062    0.01091 ± 0.00073  P < 0.05. As could be seen from Table I, the contents of gastrodin and gastrodigenin were the highest in the sample obtained by 50% ethanol extraction after boiling from G. elata, which were 0.679 and 1.125%, respectively. The content of gastrodin was the lowest in the sample obtained by 50% alcohol extraction after baking, which was ~0.108%. The content of gastrodigenin was the lowest in the sample obtained by water extraction after steaming, which was ~0.253%. Considering two indices of gastrodin and gastrodigenin (gastrodin as the main), 50% ethanol extraction after boiling was the best processing method. The 50% alcohol extraction after baking was the worst processing method for G. elata, but it is a standardized extraction method at present. This could provide reference for the processing of G. elata. Discussion The experimental results showed that the processing method had important influence on the content of gastrodin and gastrodigenin. The content of gastrodin in G. elata is not stable. Sampling at the same place and using the same processing method, the content of gastrodin measured from different individuals of G. elata is not the same, especially the different processing methods on the content of gastrodin have great influence. In order to avoid the influence of the former on the content of gastrodin and gastrodigenin, the samples were taken at the same place, and G. elata was equally divided into three groups according to their size, which ensured the ingredient of G. elata in each group was consistent. So, the results obtained in this article had a high degree of confidence when we discussed the effects of different processing methods on the content of gastrodin and gastrodigenin. In addition, boiling, steaming, baking and other initial processing methods can make the enzyme inactivated in G. elata. So, G. elata is easy to conserve. It is OK to cook thoroughly, steam thoroughly, or bake thoroughly the sample. If the initial processing time is too long, it will lead to the loss of gastrodin and gastrodigenin. Conclusion Based on the Design of Experiment (DoE) approach established by the previous researchers, the processing method of G. elata was further optimized, just like the works of Nobel Prize Winner Too Youyou on isolation of artemisinine from plants. By determining the contents of gastrodin and gastrodigenin in different samples, it indicated that 50% alcohol extraction after boiling was a reference method for processing G. elata. Funding The Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry. The work was also supported by Chongqing Key Research Project of Basic Science & Frontier Technology (No. cstc2017jcyjBX0012), Scientific and Technological Research Program of Chongqing Municipal Education Commission (Grant no. KJ1400523), Foundation Project of Chongqing Normal University (No. 14XYY020), Chongqing General Research Project of Basic Science & Frontier Technology (No. cstc2015jcyjA10054), Chongqing Normal University Postgraduate’s Research and Innovation Project (No. YKC17004), Open Foundation Project of University Engineering Research Center for Bioactive Substances in Chongqing (No. AS201614), Chongqing University Students’ Training Project of Innovation & Undertaking (No. 201610637076), China. References 1 Jeon, J.S., Kim, J., Park, S., Ryou, C., Kim, C.Y.; Preparative purification of plasmin activity stimulating phenolic derivatives from Gastrodia elata using centrifugal partition chromatography; Biomedical Chromatography , ( 2016); 30( 6): 976– 982. Google Scholar CrossRef Search ADS PubMed  2 Zhang, W., Sheng, Y.X., Zhang, J.L.; Determination and pharmacokinetics of gastrodin and p-hydroxybenzylalcohol after oral administration of Gastrodia elata Bl. extract in rats by high-performance liquid chromatography-electrospray ionization mass spectrometric method; Phytomedicine: International Journal of Phytotherapy and Phytopharmacology , ( 2008); 15( 10): 844– 850. Google Scholar CrossRef Search ADS PubMed  3 Liu, Y., Huang, G.; The chemical composition, pharmacological effects, clinical applications and market analysis of Gastrodia elata; Pharmaceutical Chemistry Journal , ( 2017); 51( 3): 211– 215. Google Scholar CrossRef Search ADS   4 Sun, W., Miao, B., Wang, X.C., Duan, J.H., Ye, X., Han, W.J., et al.  .; Gastrodin inhibits allodynia and hyperalgesia in painful diabetic neuropathy rats by decreasing excitability of nociceptive primary sensory neurons; PLoS One , ( 2012); 7( 6): e39647. Google Scholar CrossRef Search ADS PubMed  5 Guo, Z.G., Jia, X.P., Su, X.J., Li, P., Hao, J.H.; Gastrodin attenuates vincristine-induced mechanical hyperalgesia through serotonin 5-HT1A receptors; Bangladesh Journal of Pharmacology , ( 2013); 8( 4): 414– 419. Google Scholar CrossRef Search ADS   6 Li, H.B., Chen, F.; Preparative isolation and purification of gastrodin from the Chinese medicinal plant Gastrodia elata by high-speed counter-current chromatography; Journal of Chromatography A , ( 2004); 1052( 1–2): 229– 232. Google Scholar CrossRef Search ADS PubMed  7 Ong, E.S., Heng, M.Y., Tan, S.N., Hong, Y.J., Koh, H., Teo, C.C., et al.  .; Determination of gastrodin and vanillyl alcohol in Gastrodia elata Blume by pressurized liquid extraction at room temperature; Journal of Separation Science , ( 2007); 30: 2130– 2137. Google Scholar CrossRef Search ADS PubMed  © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Chromatographic Science Oxford University Press

The Content Analysis of Gastrodin and Gastrodigenin Obtained by Different Processing Methods

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Abstract

Abstract The purpose of this article was to investigate the effects of different processing methods on the content of gastrodin and gastrodigenin in Gastrodia elata Blume, so as to obtain the best processing method of G. elata. The contents of gastrodin and gastrodigenin in the samples were determined by HPLC method in turn. The results showed that the content of gastrodin and gastrodigenin were the highest in the sample obtained by extraction with 50% ethanol after boiling. Therefore, according to the two indexes of gastrodin and gastrodigenin, alcohol extraction after boiling was the best processing method, which could provide the reference for production. Introduction Gastrodia elata Blume is the subterranean dried tuber of perennial herbaceous plant in Orchidaceae (1). The gastrodin and gastrodigenin in G. elata are the main active ingredients (2), and they have specific pharmacological activities, which allow their use as medicines (3). Gastrodin has sedative, anticonvulsant, analgesic and hypnotic effects (4). Gastrodigenin has the same analgesic effect as gastrodin (5). At present, the best processing method of G. elata is based on the determination of gastrodin content (6, 7). In this study, fresh G. elata Blume was used as raw material. It was processed by water extraction after boiling, alcohol extraction after boiling, water extraction after steaming, alcohol extraction after steaming, water extraction after baking and alcohol extraction after baking, respectively. The content of gastrodin and gastrodigenin was determined by HPLC. The aim was to observe the dynamic balance of gastrodin and gastrodigenin. At the same time, it would provide a scientific and effective method for the processing of G. elata. Experimental General The samples were analyzed by Hitachi high performance liquid chromatography L-2000. Gastrodia elata (fresh) was collected from Chengkou county of Chongqing city, China. The sampling time was on 28 March 2017. Acetonitrile (MeCN) was of chromatographic-purity grade. Purified water was made at our lab. Gastrodin control, gastrodigenin control and other reagents were of analytical-reagent grade. Chromatographic condition Chromatographic column: Lachrom 891-5050; mobile phase: acetonitrile–0.05% phosphoric acid solution (3:97); detection wavelength: 220 nm; flow rate: 1.0 mL/min; column temperature: 40°C; sample size: 20 μL. The processing method of G. elata The same volume of fresh G. elata was collected, which was evenly divided into three portions. The first portion of fresh G. elata was put into boiling water and cooked thoroughly, then sliced and dried at 60°C in oven for next use. The second portion of fresh G. elata was put into the steamer and steamed thoroughly, then sliced and dried at 60°C in oven for next use. The third portion of fresh G. elata was put into the oven and heated thoroughly at 110°C, then sliced and dried at 60°C in oven for next use. Preparation of sample solution To take and crush a certain amount of the above processed sample in turn, add 50% ethanol or pure water (50 mL) and weigh. To be refluxed for 3 h and cooled. To weigh and supplement lost weight with extraction solvent. To shake and filtrate. The filtrate (20 mL) was concentrated to dryness, and 20% acetonitrile water solution was added. To be dissolved in 50 mL volumetric flask and diluted to the scale. To shake and be filtrated with microporous membrane (0.45 μm). So, the test sample solution was obtained. Results In the samples obtained by different processing methods, the content determination results of gastrodin and gastrodigenin were shown in Table I. Table I. The Content Determination Results of Gastrodin and Gastrodigenin in Samples Obtained by Different Processing Methods Sample source  Test item  Measured content (μg)  Sample size (g)  Measured content (g)/sample size (g)  Water extraction after boiling  Gastrodin  114.3359412 ± 0.0000628  0.0221  0.00517 ± 0.00063    Gastrodigenin  70.37750841 ± 0.00007323    0.00318 ± 0.00057  Alcohol extraction after boiling  Gastrodin  156.1118825 ± 0.0000195  0.023  0.00679 ± 0.00017    Gastrodigenin  258.7135519 ± 0.0000332    0.01125 ± 0.00092  Water extraction after steaming  Gastrodin  93.68121212 ± 0.0000715  0.0208  0.0045 ± 0.00068    Gastrodigenin  52.71591707 ± 0.00000746    0.00253 ± 0.00055  Alcohol extraction after steaming  Gastrodin  142.8957821 ± 0.0000162  0.0213  0.00671 ± 0.00026    Gastrodigenin  132.5645811 ± 0.0000276    0.00622 ± 0.00059  Water extraction after baking  Gastrodin  32.21564738 ± 0.00002738  0.0216  0.00149 ± 0.00085    Gastrodigenin  78.5830962 ± 0.00001835    0.00364 ± 0.00093  Alcohol extraction after baking  Gastrodin  32.10239363 ± 0.00001126  0.0298  0.00108 ± 0.00026    Gastrodigenin  325.148749 ± 0.000062    0.01091 ± 0.00073  Sample source  Test item  Measured content (μg)  Sample size (g)  Measured content (g)/sample size (g)  Water extraction after boiling  Gastrodin  114.3359412 ± 0.0000628  0.0221  0.00517 ± 0.00063    Gastrodigenin  70.37750841 ± 0.00007323    0.00318 ± 0.00057  Alcohol extraction after boiling  Gastrodin  156.1118825 ± 0.0000195  0.023  0.00679 ± 0.00017    Gastrodigenin  258.7135519 ± 0.0000332    0.01125 ± 0.00092  Water extraction after steaming  Gastrodin  93.68121212 ± 0.0000715  0.0208  0.0045 ± 0.00068    Gastrodigenin  52.71591707 ± 0.00000746    0.00253 ± 0.00055  Alcohol extraction after steaming  Gastrodin  142.8957821 ± 0.0000162  0.0213  0.00671 ± 0.00026    Gastrodigenin  132.5645811 ± 0.0000276    0.00622 ± 0.00059  Water extraction after baking  Gastrodin  32.21564738 ± 0.00002738  0.0216  0.00149 ± 0.00085    Gastrodigenin  78.5830962 ± 0.00001835    0.00364 ± 0.00093  Alcohol extraction after baking  Gastrodin  32.10239363 ± 0.00001126  0.0298  0.00108 ± 0.00026    Gastrodigenin  325.148749 ± 0.000062    0.01091 ± 0.00073  P < 0.05. As could be seen from Table I, the contents of gastrodin and gastrodigenin were the highest in the sample obtained by 50% ethanol extraction after boiling from G. elata, which were 0.679 and 1.125%, respectively. The content of gastrodin was the lowest in the sample obtained by 50% alcohol extraction after baking, which was ~0.108%. The content of gastrodigenin was the lowest in the sample obtained by water extraction after steaming, which was ~0.253%. Considering two indices of gastrodin and gastrodigenin (gastrodin as the main), 50% ethanol extraction after boiling was the best processing method. The 50% alcohol extraction after baking was the worst processing method for G. elata, but it is a standardized extraction method at present. This could provide reference for the processing of G. elata. Discussion The experimental results showed that the processing method had important influence on the content of gastrodin and gastrodigenin. The content of gastrodin in G. elata is not stable. Sampling at the same place and using the same processing method, the content of gastrodin measured from different individuals of G. elata is not the same, especially the different processing methods on the content of gastrodin have great influence. In order to avoid the influence of the former on the content of gastrodin and gastrodigenin, the samples were taken at the same place, and G. elata was equally divided into three groups according to their size, which ensured the ingredient of G. elata in each group was consistent. So, the results obtained in this article had a high degree of confidence when we discussed the effects of different processing methods on the content of gastrodin and gastrodigenin. In addition, boiling, steaming, baking and other initial processing methods can make the enzyme inactivated in G. elata. So, G. elata is easy to conserve. It is OK to cook thoroughly, steam thoroughly, or bake thoroughly the sample. If the initial processing time is too long, it will lead to the loss of gastrodin and gastrodigenin. Conclusion Based on the Design of Experiment (DoE) approach established by the previous researchers, the processing method of G. elata was further optimized, just like the works of Nobel Prize Winner Too Youyou on isolation of artemisinine from plants. By determining the contents of gastrodin and gastrodigenin in different samples, it indicated that 50% alcohol extraction after boiling was a reference method for processing G. elata. Funding The Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry. The work was also supported by Chongqing Key Research Project of Basic Science & Frontier Technology (No. cstc2017jcyjBX0012), Scientific and Technological Research Program of Chongqing Municipal Education Commission (Grant no. KJ1400523), Foundation Project of Chongqing Normal University (No. 14XYY020), Chongqing General Research Project of Basic Science & Frontier Technology (No. cstc2015jcyjA10054), Chongqing Normal University Postgraduate’s Research and Innovation Project (No. YKC17004), Open Foundation Project of University Engineering Research Center for Bioactive Substances in Chongqing (No. AS201614), Chongqing University Students’ Training Project of Innovation & Undertaking (No. 201610637076), China. References 1 Jeon, J.S., Kim, J., Park, S., Ryou, C., Kim, C.Y.; Preparative purification of plasmin activity stimulating phenolic derivatives from Gastrodia elata using centrifugal partition chromatography; Biomedical Chromatography , ( 2016); 30( 6): 976– 982. Google Scholar CrossRef Search ADS PubMed  2 Zhang, W., Sheng, Y.X., Zhang, J.L.; Determination and pharmacokinetics of gastrodin and p-hydroxybenzylalcohol after oral administration of Gastrodia elata Bl. extract in rats by high-performance liquid chromatography-electrospray ionization mass spectrometric method; Phytomedicine: International Journal of Phytotherapy and Phytopharmacology , ( 2008); 15( 10): 844– 850. Google Scholar CrossRef Search ADS PubMed  3 Liu, Y., Huang, G.; The chemical composition, pharmacological effects, clinical applications and market analysis of Gastrodia elata; Pharmaceutical Chemistry Journal , ( 2017); 51( 3): 211– 215. Google Scholar CrossRef Search ADS   4 Sun, W., Miao, B., Wang, X.C., Duan, J.H., Ye, X., Han, W.J., et al.  .; Gastrodin inhibits allodynia and hyperalgesia in painful diabetic neuropathy rats by decreasing excitability of nociceptive primary sensory neurons; PLoS One , ( 2012); 7( 6): e39647. Google Scholar CrossRef Search ADS PubMed  5 Guo, Z.G., Jia, X.P., Su, X.J., Li, P., Hao, J.H.; Gastrodin attenuates vincristine-induced mechanical hyperalgesia through serotonin 5-HT1A receptors; Bangladesh Journal of Pharmacology , ( 2013); 8( 4): 414– 419. Google Scholar CrossRef Search ADS   6 Li, H.B., Chen, F.; Preparative isolation and purification of gastrodin from the Chinese medicinal plant Gastrodia elata by high-speed counter-current chromatography; Journal of Chromatography A , ( 2004); 1052( 1–2): 229– 232. Google Scholar CrossRef Search ADS PubMed  7 Ong, E.S., Heng, M.Y., Tan, S.N., Hong, Y.J., Koh, H., Teo, C.C., et al.  .; Determination of gastrodin and vanillyl alcohol in Gastrodia elata Blume by pressurized liquid extraction at room temperature; Journal of Separation Science , ( 2007); 30: 2130– 2137. Google Scholar CrossRef Search ADS PubMed  © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

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Published: Jan 1, 2018

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