最早的中药辅料炮制品:西汉海昏侯墓出土的木质漆盒内样品鉴定与分析
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摘要
中药炮制技艺是国家级非物质文化遗产,体现了中医用药特点.有关炮制药物的文字记载最早可追溯到《黄帝内经》,但是古代对炮制工艺的记载文字简短,使复原传统技艺、实现"遵古炮制"存在一定困难.因此古代炮制药物的考古发现将为中药炮制的起源研究提供有力的实证.西汉海昏侯墓园主墓(M1)中出土了由木质漆盒盛装的样品,通过核磁及三维重建、显微分析,发现该样品是由外部辅料层和内部植物层构成.结合显微特征、质谱分析,推测该样品中为玄参科地黄属Rehmannia植物的根,且可能经过了热水等处理.《神农本草经》记载地黄具有"主折跌,绝筋,伤中.逐血痹,填骨髓,长肌肉"等功效,这与古代文史资料记载墓主生前患有严重的风湿病相符.样品的辅料层内含有淀粉粒与蔗糖,这可能与炮制"矫味矫臭、利于服用"的作用有关. M1木质漆盒内遗存样品是迄今报道的我国古代最早的中药辅料炮制品,其发现和鉴定为深入了解我国古代药物炮制与应用历史奠定了基础.
The processing of traditional Chinese medicine is an integral part of Chinese medical heritage. The recent discovery of the earliest processed drugs may provide strong evidence for the origins of traditional Chinese medicine processing. The remains were found in wooden lacquer boxes, unearthed from Haihunhou tomb in Nanchang, Jiangxi Province. By analyzing the appearance of the unearthed samples, it was found that the outer layers of the samples were composed of an auxiliary material and the inside was composed of plant material. As can be seen from the three-dimensional image taken using nuclear magnetic resonance screening,the sample had a hollow irregular rod structure. The outer layers were composed of a substance with weak signal strength, and the inside was similar to the fiber structure of plants with high water contents. Microscopic analysis showed that periderm, secondary xylem, and broad wood rays could be seen in the transverse section of the plants inside the samples. It can be further speculated that the plants inside the samples were derived from the roots or stems of dicotyledonous herbs. Additionally, the putative roots or stems of the dicotyledonous herbs had secondary structures and contained one to three clustered, red-brown cells. No starch granules, calcium oxalate crystals, or stone cells were observed in the transverse section. According to the microscopic database of the National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, it is speculated that the plants inside the samples were derived from the roots of Rehmannia species. A comparative study of the microscopic characteristics of wild Rehmannia chingii roots and the unearthed samples was carried out. It was found that both displayed periderm, broad secondary phloem, wood rays, similar vessel morphologies, and red-brown cells. The unearthed samples and R.chingii roots were analyzed by electrospray ionization mass spectrometry(ESI-MS) in negative ion mode. The results showed that the mass spectrum peak of m/z 623 in the unearthed samples agreed with that in the roots of R. chingii, and was putatively identified as acteoside or forsythiaside. Moreover, the peak at m/z 179 in the unearthed samples was identified as caffeic acid based on the molecular weight using high resolution tandem mass spectrometry(MS/MS) data, and was usually identified as a hydrolysate of acteoside or forsythiaside. There were about two to three outer layers of excipient on the samples. They were distributed unevenly, thicker at the top and thinner at the bottom. The plants and excipient layers were observed by microscope,and starch grains were found in the cross section and powder of the excipient layers, while no starch grains were found in the cross section or powder of the plant material inside. However, the reaction between the plant tissues and iodine-potassium iodide test solution produced the characteristic blue color. This phenomenon was related to the gelatinization of starch granules. Under a polarized light microscope, a large number of crystals were seen in the excipient layers. An aqueous extract of the excipient layers was analyzed by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOFMS), and an ion fragment peak at m/z 341.11 was detected, indicating a disaccharide. When compared to the reference substances glucose, sucrose, fructose, and maltose, the retention times and mass spectra were very similar to that of sucrose. According to ancient literary and historical records, the tomb owner suffered from severe rheumatism before his death, which is consistent with the efficacy of R. chingii. It is further speculated that the samples in the wooden lacquer boxes are the remains of processed ancient traditional Chinese medicine products. The origin of the excipients in the processed medicines might be related to improving the taste, which could provide a scientific basis for further understanding the processing and application of ancient traditional Chinese medicines.
The processing of traditional Chinese medicine is an integral part of Chinese medical heritage. The recent discovery of the earliest processed drugs may provide strong evidence for the origins of traditional Chinese medicine processing. The remains were found in wooden lacquer boxes, unearthed from Haihunhou tomb in Nanchang, Jiangxi Province. By analyzing the appearance of the unearthed samples, it was found that the outer layers of the samples were composed of an auxiliary material and the inside was composed of plant material. As can be seen from the three-dimensional image taken using nuclear magnetic resonance screening,the sample had a hollow irregular rod structure. The outer layers were composed of a substance with weak signal strength, and the inside was similar to the fiber structure of plants with high water contents. Microscopic analysis showed that periderm, secondary xylem, and broad wood rays could be seen in the transverse section of the plants inside the samples. It can be further speculated that the plants inside the samples were derived from the roots or stems of dicotyledonous herbs. Additionally, the putative roots or stems of the dicotyledonous herbs had secondary structures and contained one to three clustered, red-brown cells. No starch granules, calcium oxalate crystals, or stone cells were observed in the transverse section. According to the microscopic database of the National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, it is speculated that the plants inside the samples were derived from the roots of Rehmannia species. A comparative study of the microscopic characteristics of wild Rehmannia chingii roots and the unearthed samples was carried out. It was found that both displayed periderm, broad secondary phloem, wood rays, similar vessel morphologies, and red-brown cells. The unearthed samples and R.chingii roots were analyzed by electrospray ionization mass spectrometry(ESI-MS) in negative ion mode. The results showed that the mass spectrum peak of m/z 623 in the unearthed samples agreed with that in the roots of R. chingii, and was putatively identified as acteoside or forsythiaside. Moreover, the peak at m/z 179 in the unearthed samples was identified as caffeic acid based on the molecular weight using high resolution tandem mass spectrometry(MS/MS) data, and was usually identified as a hydrolysate of acteoside or forsythiaside. There were about two to three outer layers of excipient on the samples. They were distributed unevenly, thicker at the top and thinner at the bottom. The plants and excipient layers were observed by microscope,and starch grains were found in the cross section and powder of the excipient layers, while no starch grains were found in the cross section or powder of the plant material inside. However, the reaction between the plant tissues and iodine-potassium iodide test solution produced the characteristic blue color. This phenomenon was related to the gelatinization of starch granules. Under a polarized light microscope, a large number of crystals were seen in the excipient layers. An aqueous extract of the excipient layers was analyzed by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOFMS), and an ion fragment peak at m/z 341.11 was detected, indicating a disaccharide. When compared to the reference substances glucose, sucrose, fructose, and maltose, the retention times and mass spectra were very similar to that of sucrose. According to ancient literary and historical records, the tomb owner suffered from severe rheumatism before his death, which is consistent with the efficacy of R. chingii. It is further speculated that the samples in the wooden lacquer boxes are the remains of processed ancient traditional Chinese medicine products. The origin of the excipients in the processed medicines might be related to improving the taste, which could provide a scientific basis for further understanding the processing and application of ancient traditional Chinese medicines.
引文
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2 Li W.Huangdi Neijing(in Chinese).Shenyang:Liaoning Ethnic Publishing House,1999[李文.黄帝内经.沈阳:辽宁民族出版社,1999]
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4 Zhang Z J.Annotation of Shang Han Lun(in Chinese).Changsha:Hunan Science and Technology Press,1982[张仲景.伤寒论校注.长沙:湖南科学技术出版社,1982]
5 He R.Annotation of Jinkui Yaolue(in Chinese).Beijing:People’s Medical Publishing House,1990[何任.《金匮要略》校注.北京:人民卫生出版社,1990]
6 Huang L Q,Sun L Y,Zhang X B,et al.Brief introduction to progress of national census of Chinese medicine resources(pilot)(in Chinese).China JChin Mater Med,2017,42:4256-4261[黄璐琦,孙丽英,张小波,等.全国中药资源普查(试点)工作进展情况简介.中国中药杂志,2017,42:4256-4261]
7 Jiang T X.Flora Dictionary(in Chinese).(Photocopy Edition).Volume I.Shanghai:Shanghai Literature&Art Publishing House,1998[蒋廷锡.草木典.(影印版).上册.上海:上海文艺出版社,1998]
8 Compilation Committee of Chinese flora of the Chinese Academy of Sciences.Flora Reipublicae Popularis Sinicae(in Chinese).Beijing:Science Press,1979.67[中国科学院中国植物志编辑委员会.中国植物志.北京:科学出版社,1979.67]
9 Tang S W.Da guan Ben cao(in Chinese).Shang Z J collated.Hefei:Anhui Science&Technology Press,2002.180-181[唐慎微.大观本草.尚志钧点校.合肥:安徽科学技术出版社,2002.180-181]
10 Liu M Q,Wang X Q,Chen S Q,et al.Histochemistry of Rehmannia glutinosa(in Chinese).J Chin Med Mater,2013,36:1771-1773[刘孟奇,王小巧,陈随清,等.地黄的组织化学研究.中药材,2013,36:1771-1773]
11 Wu Z C,Luo G M,Huo Y C.Extraction,isolation and TLC quantification of secretions from Rehmannia glutinosa secretory cells(in Chinese).Chin J Pharm Anal(Suppl),1998,18:248-250[吴子超,罗干明,霍永昌.地黄分泌细胞分泌物的提取、分离和薄层扫描定量.药物分析杂志(增刊),1998,18:248-250]
12 Zhang B Y,Jiang Z Z,Wang Y F,et al.Analysis of chemical constituents in fresh,dried and prepared Rehmannniae Radix by UPLC/ESI-Q-TOFMS(in Chinese).Chin Trad Patent Med,2016,38:1104-1108[张波泳,江振作,王跃飞,等.UPLC/ESI-Q-TOF MS法分析鲜地黄、生地黄、熟地黄的化学成分.中成药,2016,38:1104-1108]
13 Meng X Y.Studies on chemistry composition,standard of quality and anti-senescence activities of Rehmannia chingii Li(in Chinese).Master Dissertation.Yinchuan:Ningxia Medical University,2015[蒙雄裕.天目地黄化学成分、治疗标准及其抗衰老活性研究.硕士学位论文.银川:宁夏医科大学,2015]
14 Li S L,Song J Z,Qiao C F,et al.A novel strategy to rapidly explore potential chemical markers for the discrimination between raw and processed Radix Rehmanniae by UHPLC-TOFMS with multivariate statistical analysis.J Pharm Biomed Anal,2010,51:812-823
15 Qi M,Xiong A,Li P,et al.Identification of acteoside and its major metabolites in rat urine by ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.J Chromatogr B,2013,940:77-85
16 Zeng G,Xiao H,Liu J,et al.Identification of phenolic constituents in RadixSalvia miltiorrhizae by liquid chromatography/electrospray ionization mass spectrometry.Rapid Commun Mass Spectrom,2006,20:499-506
17 Geng D D,Dong Q,Tan L,et al.On-line screening and identification of free adical scavenging compounds in Salvia miltiorrhiza Bunge and Salvia prattii Hemsl.by HPLC-DAD-ESI/MSn-DPPH(in Chinese).J Instrum Anal,2015,34:314-320[耿丹丹,董琦,谭亮,等.HPLC-DAD-ESI/MSnDPPH在线筛选与鉴别丹参和康定鼠尾草中抗氧化活性成分.分析测试学报,2015,34:314-320]
18 Pharmacopoeia of the People’s Republic of China.Pharmacopoeia Commission of the People’s Republic of China(in Chinese).Beijing:China Medical Science Press,2015.124[国家药典委员会.中华人民共和国药典.一部.北京:中国医药科技出版社,2015.124]
19 Kang T G.Authentication of traditional Chinese medicine(in Chinese).Beijing:China Press of Traditional Chinese Medicine,2016.152-153[康廷国.中药鉴定学.北京:中国中医药出版社,2016.152-153]
20 Hu Z H.The relationship between the structure,development and medicinal components of medicinal plants(in Chinese).Shanghai:Shanghai Science and Technology Press,2014.352-354[胡正海.药用植物的结构、发育与药用成分的关系.上海:上海科学技术出版社,2014.352-354]
21 Cheng Y L,Mei L T,Kuo H T.Effect of amylose content on the rheological property of rice starch.Cereal Chem,1996,73:415-420
22 Ban G.History of the Han Dynasty(in Chinese).Beijing:Chinese Publishing House,2016[班固.汉书.北京:中华书局,2016]
23 Xu S.Origin of Chinese Characters(in Chinese).Beijing:Chinese Publishing House,1985[许慎.说文解字.北京:中华书局,1985]
24 Unknown.Plain Questions of Yellow Emperor’s Canon of Medicine(in Chinese).Beijing:China Medical Science and Technology Press,2018[佚名.黄帝内经·素问.北京:中国医药科技出版社,2018]
25 Sun X Y.The Shen Nong Ben Cao Jin(in Chinese).Beijing:People’s Medical Publishing House,1963.13[孙星衍辑.神农本草经.北京:人民卫生出版社,1963.13]
26 Lei X.Lei Gong Pao Zhi Lun(in Chinese).Hefei:Anhui Science and Technology Publishing House,1991.24[雷敩.雷公炮炙论.合肥:安徽科学技术出版社,1991.24]
27 Jiangxi Institute of Archaeology,Nanchang museum,Nanchang Xinjian District museum.Haihunhou Han Tomb(in Chinese).Archaeol,2016,(7):45-62[江西省文物考古研究所,南昌市博物馆,南昌市新建区博物馆.南昌市西汉海昏侯墓.考古,2016,(7):45-62]
28 Ji Q L.The History of Sucrose(in Chinese).Beijing:China Customs Publishing House,2009[季羡林.蔗糖史.北京:中国海关出版社,2009]
29 Zhang L.A Variorum of Prescriptions of Fifty-two Diseases of Mawangdui Han Tomb(in Chinese).Beijing:Ancient Books Publishing House of Traditional Chinese Medicine,2017[张雷.马王堆汉墓帛书《五十二病方》集注.北京:中医古籍出版社,2017]
30 Zhang Z J.Typhus Various Illnesses to Discuss(Guilin Guben)(in Chinese).Beijing:China Press of Traditional Chinese Medicine,2014[张仲景.伤寒杂病论(桂林古本).北京:中国中医药出版社,2014]
31 Li S Z.Compendium of Materia Medica(in Chinese).Volume I.Beijing:Huaxia Publishing House,1998.706[李时珍.本草纲目.上册.北京:华夏出版社,1998.706]
2 Li W.Huangdi Neijing(in Chinese).Shenyang:Liaoning Ethnic Publishing House,1999[李文.黄帝内经.沈阳:辽宁民族出版社,1999]
3 Shang Z Z.Notes on Medicine in“Fifty-two Patient Prescriptions”(in Chinese).Wuhu:Scientific Research Center of Wannan Medical College,1985[尚志钧.《五十二病方》药物注释.芜湖:皖南医学院科研科,1985]
4 Zhang Z J.Annotation of Shang Han Lun(in Chinese).Changsha:Hunan Science and Technology Press,1982[张仲景.伤寒论校注.长沙:湖南科学技术出版社,1982]
5 He R.Annotation of Jinkui Yaolue(in Chinese).Beijing:People’s Medical Publishing House,1990[何任.《金匮要略》校注.北京:人民卫生出版社,1990]
6 Huang L Q,Sun L Y,Zhang X B,et al.Brief introduction to progress of national census of Chinese medicine resources(pilot)(in Chinese).China JChin Mater Med,2017,42:4256-4261[黄璐琦,孙丽英,张小波,等.全国中药资源普查(试点)工作进展情况简介.中国中药杂志,2017,42:4256-4261]
7 Jiang T X.Flora Dictionary(in Chinese).(Photocopy Edition).Volume I.Shanghai:Shanghai Literature&Art Publishing House,1998[蒋廷锡.草木典.(影印版).上册.上海:上海文艺出版社,1998]
8 Compilation Committee of Chinese flora of the Chinese Academy of Sciences.Flora Reipublicae Popularis Sinicae(in Chinese).Beijing:Science Press,1979.67[中国科学院中国植物志编辑委员会.中国植物志.北京:科学出版社,1979.67]
9 Tang S W.Da guan Ben cao(in Chinese).Shang Z J collated.Hefei:Anhui Science&Technology Press,2002.180-181[唐慎微.大观本草.尚志钧点校.合肥:安徽科学技术出版社,2002.180-181]
10 Liu M Q,Wang X Q,Chen S Q,et al.Histochemistry of Rehmannia glutinosa(in Chinese).J Chin Med Mater,2013,36:1771-1773[刘孟奇,王小巧,陈随清,等.地黄的组织化学研究.中药材,2013,36:1771-1773]
11 Wu Z C,Luo G M,Huo Y C.Extraction,isolation and TLC quantification of secretions from Rehmannia glutinosa secretory cells(in Chinese).Chin J Pharm Anal(Suppl),1998,18:248-250[吴子超,罗干明,霍永昌.地黄分泌细胞分泌物的提取、分离和薄层扫描定量.药物分析杂志(增刊),1998,18:248-250]
12 Zhang B Y,Jiang Z Z,Wang Y F,et al.Analysis of chemical constituents in fresh,dried and prepared Rehmannniae Radix by UPLC/ESI-Q-TOFMS(in Chinese).Chin Trad Patent Med,2016,38:1104-1108[张波泳,江振作,王跃飞,等.UPLC/ESI-Q-TOF MS法分析鲜地黄、生地黄、熟地黄的化学成分.中成药,2016,38:1104-1108]
13 Meng X Y.Studies on chemistry composition,standard of quality and anti-senescence activities of Rehmannia chingii Li(in Chinese).Master Dissertation.Yinchuan:Ningxia Medical University,2015[蒙雄裕.天目地黄化学成分、治疗标准及其抗衰老活性研究.硕士学位论文.银川:宁夏医科大学,2015]
14 Li S L,Song J Z,Qiao C F,et al.A novel strategy to rapidly explore potential chemical markers for the discrimination between raw and processed Radix Rehmanniae by UHPLC-TOFMS with multivariate statistical analysis.J Pharm Biomed Anal,2010,51:812-823
15 Qi M,Xiong A,Li P,et al.Identification of acteoside and its major metabolites in rat urine by ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.J Chromatogr B,2013,940:77-85
16 Zeng G,Xiao H,Liu J,et al.Identification of phenolic constituents in RadixSalvia miltiorrhizae by liquid chromatography/electrospray ionization mass spectrometry.Rapid Commun Mass Spectrom,2006,20:499-506
17 Geng D D,Dong Q,Tan L,et al.On-line screening and identification of free adical scavenging compounds in Salvia miltiorrhiza Bunge and Salvia prattii Hemsl.by HPLC-DAD-ESI/MSn-DPPH(in Chinese).J Instrum Anal,2015,34:314-320[耿丹丹,董琦,谭亮,等.HPLC-DAD-ESI/MSnDPPH在线筛选与鉴别丹参和康定鼠尾草中抗氧化活性成分.分析测试学报,2015,34:314-320]
18 Pharmacopoeia of the People’s Republic of China.Pharmacopoeia Commission of the People’s Republic of China(in Chinese).Beijing:China Medical Science Press,2015.124[国家药典委员会.中华人民共和国药典.一部.北京:中国医药科技出版社,2015.124]
19 Kang T G.Authentication of traditional Chinese medicine(in Chinese).Beijing:China Press of Traditional Chinese Medicine,2016.152-153[康廷国.中药鉴定学.北京:中国中医药出版社,2016.152-153]
20 Hu Z H.The relationship between the structure,development and medicinal components of medicinal plants(in Chinese).Shanghai:Shanghai Science and Technology Press,2014.352-354[胡正海.药用植物的结构、发育与药用成分的关系.上海:上海科学技术出版社,2014.352-354]
21 Cheng Y L,Mei L T,Kuo H T.Effect of amylose content on the rheological property of rice starch.Cereal Chem,1996,73:415-420
22 Ban G.History of the Han Dynasty(in Chinese).Beijing:Chinese Publishing House,2016[班固.汉书.北京:中华书局,2016]
23 Xu S.Origin of Chinese Characters(in Chinese).Beijing:Chinese Publishing House,1985[许慎.说文解字.北京:中华书局,1985]
24 Unknown.Plain Questions of Yellow Emperor’s Canon of Medicine(in Chinese).Beijing:China Medical Science and Technology Press,2018[佚名.黄帝内经·素问.北京:中国医药科技出版社,2018]
25 Sun X Y.The Shen Nong Ben Cao Jin(in Chinese).Beijing:People’s Medical Publishing House,1963.13[孙星衍辑.神农本草经.北京:人民卫生出版社,1963.13]
26 Lei X.Lei Gong Pao Zhi Lun(in Chinese).Hefei:Anhui Science and Technology Publishing House,1991.24[雷敩.雷公炮炙论.合肥:安徽科学技术出版社,1991.24]
27 Jiangxi Institute of Archaeology,Nanchang museum,Nanchang Xinjian District museum.Haihunhou Han Tomb(in Chinese).Archaeol,2016,(7):45-62[江西省文物考古研究所,南昌市博物馆,南昌市新建区博物馆.南昌市西汉海昏侯墓.考古,2016,(7):45-62]
28 Ji Q L.The History of Sucrose(in Chinese).Beijing:China Customs Publishing House,2009[季羡林.蔗糖史.北京:中国海关出版社,2009]
29 Zhang L.A Variorum of Prescriptions of Fifty-two Diseases of Mawangdui Han Tomb(in Chinese).Beijing:Ancient Books Publishing House of Traditional Chinese Medicine,2017[张雷.马王堆汉墓帛书《五十二病方》集注.北京:中医古籍出版社,2017]
30 Zhang Z J.Typhus Various Illnesses to Discuss(Guilin Guben)(in Chinese).Beijing:China Press of Traditional Chinese Medicine,2014[张仲景.伤寒杂病论(桂林古本).北京:中国中医药出版社,2014]
31 Li S Z.Compendium of Materia Medica(in Chinese).Volume I.Beijing:Huaxia Publishing House,1998.706[李时珍.本草纲目.上册.北京:华夏出版社,1998.706]