~(60)Co-γ射线辐照对中药化学成分影响及辐照中药鉴定的热释光分析法研究
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摘要
60Co-γ射线辐照灭菌法，具有穿透力强、灭菌效果好、成本低廉及对大规模生产实用性高等特点，是一种新型、高效的药品灭菌方法，近年来在中药灭菌中得到越来越广泛的应用。
本论文对中药辐照灭菌进行了系统的研究，主要包括以下几个方面：
1、对样品分别进行了不同剂量的辐照，确定60Co-γ射线辐照具有较好的灭菌效果，将样品储存3个月、6个月、9个月及18个月后，检测样品中的微生物数量。确定60Co-γ射线辐照灭菌能延长中药的保质期，且辐照剂量越大灭菌效果越好。
2、利用高效液相色谱法对人参、西洋参、三七、甘草及五味子进行不同剂量的辐照，考察中药化学成分的变化。确定经过不同剂量60Co-γ射线辐照的中药没有新物质的生成和原有成分的消失，但某些化学物质的含量出现变化。
3、通过铁离子还原能力法(FRAP)和1,1-二苯基-2-三硝基苯肼自由基清除法(DPPH)两种方法，对不同剂量辐照的5种中药抗氧化活性进行了评价。确定辐照对不同中药的抗氧化活性有不同的影响。
4、利用热释光分析法对辐照中药进行检测鉴别，对热释光发光曲线、发光强度和TL比进行研究，确定了热释光法能对混入不同比例辐照样品、储存不同时间的辐照样品及对样品进行盲检得出正确的判定。
本论文的研究结果为辐照对中药化学成分影响的研究、国家辐照灭菌标准的制定奠定了基础。
Herbs, especially Chinese herbs, have been essential in diets, health supplements, andtraditional medicines since ancient times. It is still a common practice in many parts of theworld to collect herbs and store them dried for long periods. In recent years, irradiation oftraditional Chinese herbs with60Co-γrays has been a predominant sterilization methodduring storage and transport against microbial deterioration or insect infestation in order tomaintain hygienic quality, ensure shelf life, and thereby reduce the threat to public health.Unlike other sterilization methods such as fumigation and thermal treatment, these methodscan pose occupational health hazard with environment ally destructive toxic chemicals,irradiation technique is very fast, cost effective, and provides a wild range of disinfectingapplications for dry ingredients.
In this paper, we have studied on the chemical composition and analytical techniquesfor identifying irradiated or non-irradiated, including four aspects:
1、 The study of sterilization effect by60Co-γ ray irradiation
The samples were irradiated with different doses respectively. The results showed that60Co-γ-ray irradiation has a good effect on sterilization efficiency, and the higher dose ofradiation the better sterilization effect. The initial number of colonies in the sample effectimpact on irradiation sterilization, the number of colonies in the initial sample is higher, theirradiation dose for greater sterilization is more.
The samples were stored for3months,6months,9months and18months in order toexamine the effects of irradiation on the microbiological. Irradiation herbs have a muchlonger shelf life. The total bacterial populations of irradiated
2、Effect of irradiation on the chemical composition of traditional Chinese medicine
The main chromatographic fingerprint peaks of ginseng, Panax quinquefolius, Panaxnotoginseng FH chen, licorice and schisandra were established for evaluating the stabilityof chemical compositions of Rhizoma gastrodiae with60Co-γ irradiation to control the dosage of60Co-γ irradiation sterilization by high performance liquid chromatography(HPLC). The results show that radiation dose levels have different effects on ginsenosides.Below8kGy, irradiation has little effect on them. Ginsenoside Re and Rd in irradiated at thedose of10kGy, saponin content began to decrease. After irradiation of Panax quinquefolius,four kinds of saponin content changes and the ginsenoside Rg2content increased. Panaxnotoginseng FH chen after irradiation with two kinds of saponin content change. Irradiationpromoted in licorice of liquiritin and glycyrrhizicacid in chemical reaction, which make itscontent to have the change,and with the increase of irradiation dose chemical componentcontent decreased. The composition structure of same original Schisandra in differentradiation dose was same, possibly due to the lignan compound belongs biphenyl cyclooctene,which helps to increase the stability of the benzene ring of anti-radiation.
Studies on the chemical constituents of different irradiation doses of ginseng, Americanginseng, Panax, licorice and Schisandra by using high performance liquid ChromatographChanges in composition of dried ginseng, American ginseng, Panax, licorice and schisan andin composition of the irradiated samples chemical composition were analyzed. The resultsshowed that low doses of radiation has no significant effect on the chemical composition offive Chinese medicine ingredients and high dose irradiation causes a change in medicine.Dried herbs has a good ability of anti-radiation,almost no change in the chemicalcomposition of the content. The chemical composition of the irradiated medicine afterstorage at different times did not change significantly.
3、Antioxidant activity of irradiated Chinese herbs
The antioxidant activity of different irradiated doses Chinese herbal medicines had beensuccessfully studied by the ferric reducing antioxidant power (FRAP) and radical scavenging(DPPH) methods. The results showed that low doses of radiation has no effect on antioxidantactivity of five kinds of traditional Chinese medicine. When the radiation dose is greater than5kGy, the antioxidant activity of five kinds of Chinese medicine have increased slightly.
4、The identification of irradiated herbs by TL analytical methods
The feasibility of thermoluminescence (TL) to differentiate irradiated Chinese medicinalherbs from non-irradiated was investigated. Thirty different dried Chinese herbs were tested, including root, flower, ramulus, rhizome, cortex, and whole plant samples. Irradiation ofChinese herbs was associated with strong TL peaks at~150-250°C, while TL curves ofnon-irradiated herbs had very low intensities above250°C, which was also confirmed by theTL ratio (non-irradiated, TL1/TL2<0.1). The ability to determine the irradiation dose by theTL method was influenced by the amount and types of minerals in the samples. All levels ofirradiation doses could be detected when between0.1and1.0kGy, except for three herbs at0.1kGy dose. Different blends with small quantities (0.1-10%) of irradiated herbs were alsotested in this study. Samples with powder mixtures containing1%irradiated componentscould be differentiated (TL1/TL2>0.1) except for sterculialychnophora, semen cassia,flosinulae, and anemone root. TL ratios of some herbs indicated irradiation (TL1/TL2>0.1)even if the irradiated components were as low as0.1%. The shape of thermoluminescenceglow curve shape had no obvious change with the storage time and the intensity ofthermoluminescence with the storage time has the tendency to weaken, even if the18months storage time for the irradiated samples, its intensity is far higher than that ofnon-irradiated samples. Sample blind detection results show that irradiation dose, which isused in1kGy after storage for18months, the accuracy rate of TL method is still100%.
本论文对中药辐照灭菌进行了系统的研究，主要包括以下几个方面：
1、对样品分别进行了不同剂量的辐照，确定60Co-γ射线辐照具有较好的灭菌效果，将样品储存3个月、6个月、9个月及18个月后，检测样品中的微生物数量。确定60Co-γ射线辐照灭菌能延长中药的保质期，且辐照剂量越大灭菌效果越好。
2、利用高效液相色谱法对人参、西洋参、三七、甘草及五味子进行不同剂量的辐照，考察中药化学成分的变化。确定经过不同剂量60Co-γ射线辐照的中药没有新物质的生成和原有成分的消失，但某些化学物质的含量出现变化。
3、通过铁离子还原能力法(FRAP)和1,1-二苯基-2-三硝基苯肼自由基清除法(DPPH)两种方法，对不同剂量辐照的5种中药抗氧化活性进行了评价。确定辐照对不同中药的抗氧化活性有不同的影响。
4、利用热释光分析法对辐照中药进行检测鉴别，对热释光发光曲线、发光强度和TL比进行研究，确定了热释光法能对混入不同比例辐照样品、储存不同时间的辐照样品及对样品进行盲检得出正确的判定。
本论文的研究结果为辐照对中药化学成分影响的研究、国家辐照灭菌标准的制定奠定了基础。
Herbs, especially Chinese herbs, have been essential in diets, health supplements, andtraditional medicines since ancient times. It is still a common practice in many parts of theworld to collect herbs and store them dried for long periods. In recent years, irradiation oftraditional Chinese herbs with60Co-γrays has been a predominant sterilization methodduring storage and transport against microbial deterioration or insect infestation in order tomaintain hygienic quality, ensure shelf life, and thereby reduce the threat to public health.Unlike other sterilization methods such as fumigation and thermal treatment, these methodscan pose occupational health hazard with environment ally destructive toxic chemicals,irradiation technique is very fast, cost effective, and provides a wild range of disinfectingapplications for dry ingredients.
In this paper, we have studied on the chemical composition and analytical techniquesfor identifying irradiated or non-irradiated, including four aspects:
1、 The study of sterilization effect by60Co-γ ray irradiation
The samples were irradiated with different doses respectively. The results showed that60Co-γ-ray irradiation has a good effect on sterilization efficiency, and the higher dose ofradiation the better sterilization effect. The initial number of colonies in the sample effectimpact on irradiation sterilization, the number of colonies in the initial sample is higher, theirradiation dose for greater sterilization is more.
The samples were stored for3months,6months,9months and18months in order toexamine the effects of irradiation on the microbiological. Irradiation herbs have a muchlonger shelf life. The total bacterial populations of irradiated
2、Effect of irradiation on the chemical composition of traditional Chinese medicine
The main chromatographic fingerprint peaks of ginseng, Panax quinquefolius, Panaxnotoginseng FH chen, licorice and schisandra were established for evaluating the stabilityof chemical compositions of Rhizoma gastrodiae with60Co-γ irradiation to control the dosage of60Co-γ irradiation sterilization by high performance liquid chromatography(HPLC). The results show that radiation dose levels have different effects on ginsenosides.Below8kGy, irradiation has little effect on them. Ginsenoside Re and Rd in irradiated at thedose of10kGy, saponin content began to decrease. After irradiation of Panax quinquefolius,four kinds of saponin content changes and the ginsenoside Rg2content increased. Panaxnotoginseng FH chen after irradiation with two kinds of saponin content change. Irradiationpromoted in licorice of liquiritin and glycyrrhizicacid in chemical reaction, which make itscontent to have the change,and with the increase of irradiation dose chemical componentcontent decreased. The composition structure of same original Schisandra in differentradiation dose was same, possibly due to the lignan compound belongs biphenyl cyclooctene,which helps to increase the stability of the benzene ring of anti-radiation.
Studies on the chemical constituents of different irradiation doses of ginseng, Americanginseng, Panax, licorice and Schisandra by using high performance liquid ChromatographChanges in composition of dried ginseng, American ginseng, Panax, licorice and schisan andin composition of the irradiated samples chemical composition were analyzed. The resultsshowed that low doses of radiation has no significant effect on the chemical composition offive Chinese medicine ingredients and high dose irradiation causes a change in medicine.Dried herbs has a good ability of anti-radiation,almost no change in the chemicalcomposition of the content. The chemical composition of the irradiated medicine afterstorage at different times did not change significantly.
3、Antioxidant activity of irradiated Chinese herbs
The antioxidant activity of different irradiated doses Chinese herbal medicines had beensuccessfully studied by the ferric reducing antioxidant power (FRAP) and radical scavenging(DPPH) methods. The results showed that low doses of radiation has no effect on antioxidantactivity of five kinds of traditional Chinese medicine. When the radiation dose is greater than5kGy, the antioxidant activity of five kinds of Chinese medicine have increased slightly.
4、The identification of irradiated herbs by TL analytical methods
The feasibility of thermoluminescence (TL) to differentiate irradiated Chinese medicinalherbs from non-irradiated was investigated. Thirty different dried Chinese herbs were tested, including root, flower, ramulus, rhizome, cortex, and whole plant samples. Irradiation ofChinese herbs was associated with strong TL peaks at~150-250°C, while TL curves ofnon-irradiated herbs had very low intensities above250°C, which was also confirmed by theTL ratio (non-irradiated, TL1/TL2<0.1). The ability to determine the irradiation dose by theTL method was influenced by the amount and types of minerals in the samples. All levels ofirradiation doses could be detected when between0.1and1.0kGy, except for three herbs at0.1kGy dose. Different blends with small quantities (0.1-10%) of irradiated herbs were alsotested in this study. Samples with powder mixtures containing1%irradiated componentscould be differentiated (TL1/TL2>0.1) except for sterculialychnophora, semen cassia,flosinulae, and anemone root. TL ratios of some herbs indicated irradiation (TL1/TL2>0.1)even if the irradiated components were as low as0.1%. The shape of thermoluminescenceglow curve shape had no obvious change with the storage time and the intensity ofthermoluminescence with the storage time has the tendency to weaken, even if the18months storage time for the irradiated samples, its intensity is far higher than that ofnon-irradiated samples. Sample blind detection results show that irradiation dose, which isused in1kGy after storage for18months, the accuracy rate of TL method is still100%.
引文
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