参麦注射液组方药材浙麦冬的质量控制方法研究
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摘要
质量控制水平低下是制约中药产业发展的技术瓶颈,为了提高中药质量控制水平,制造出安全、有效、稳定和质量可控的现代中药,需要充分运用现代科学的新理论和新技术,进一步加强对中药质量控制涉及到的三个关键环节—化学物质基础、质量分析方法和生产工艺的研究。
参麦注射液由红参和麦冬2味中药组成,是中药大品种。目前参麦注射液的质量控制仍存在不足之处—对组方药材浙麦冬的质量控制研究较少。鉴于此,本论文以浙麦冬为研究对象,开展其化学物质基础、质量分析方法和生产工艺的研究,希望通过本研究可以全面提高浙麦冬的质量控制水平,并为中药质量控制方法研究提供一个良好的范例。
本论文主要研究内容及学术贡献如下：
1.采用多种色谱技术系统研究浙麦冬的化学成分,分离得到12个化合物。综合运用质谱和核磁共振等波谱技术鉴定了11个化合物的结构,其中化合物8-10为新的高异黄酮类化合物,化合物1为新的甾体皂苷类化合物,化合物5为属内首次分离的甾体皂苷类化合物。
2.以6个甾体皂苷类化合物的多级质谱裂解规律为基础建立浙麦冬甾体皂苷类化学成分的HPLC-MSn定性方法。本方法可以在没有对照品的情况下快速辨识麦冬及其制剂中的甾体皂苷类化合物,初步解决了此类化合物分离难度大、结构鉴定困难的问题。建立浙麦冬药材的HPLC-MS指纹图谱,在HPLC-MSn定性分析方法的基础上并结合文献鉴定或推测了指纹图谱谱峰对应的化合物结构。
3.利用质谱的高灵敏度和高选择性建立同时测定浙麦冬3个甾体皂苷化合物的HPLC-MS定量分析方法,初步解决了麦冬甾体皂苷化合物由于含量低、紫外末端吸收,含量测定困难的问题。本方法相对于以往的分析方法具有样品预处理简单,节省分析时间等优势,可以用来区分不同产地的麦冬类药材和评价工艺中的麦冬中间体。本方法为提高浙麦冬质量控水平提供简单可行的途径。
4.实验室模拟浙麦冬4个操作单元的生产过程,每个操作单元设置12个正常批次和若干异常批次。利用HPLC-MS指纹图谱结合多变量数据分析技术辨识指纹图谱中与异常批次工艺参数变化相关的化合物,确立了单元操作中影响浙麦冬中间体质量的关键工艺参数。本研究为浙麦冬的工艺评价方法提供新的思路。
5.探索性研究了膜分离技术在浙麦冬生产中的应用,考察陶瓷膜和超滤膜对3个甾体皂苷通过率和总固含物清除率的影响,筛选出最佳孔径的陶瓷膜和最佳截留分子量的超滤膜以及合适的膜分离工艺参数,并初步研究了膜芯材料的污染问题。本研究既可以为参麦注射液的超滤工艺提供研究思路,又可以作为新工艺对参麦注射液工艺进行改进；研究活性炭吸附工艺过程中3个甾体皂苷的损失情况,探讨活性炭对3个甾体皂苷的吸附机理。
The quality control level of traditional Chinese medicine (TCM) industry is low and restricts its development. In order to improve the quality control level of TCM, the research on three critical units—hemical material basis, analytical methods and production processes in TCM quality control should be strengthed via modern science and technology.
Shenmai Injection is one of the major TCM composed of Panax ginseng (Hongshen) and Ophiopogon japonicus (Maidong). However, the quality control of Shenmai Injection is still inadequacies because the research on O. japonicus is less. In this thesis, chemical material basis, analytical methods and production processes for O. japonicus were studied. This study will raise the quality control level of O. japonicus and provide a good example for TCM quality control.
The main innovation and academic contributions of this dissertation are summarized as follows:
1. Column chromatography techniques were used to isolate the chemical constituents of O. japonicus grown in Zhejiang province.12compounds were obtained and11of them were identified via MS and NMR spectral analysis. Compounds8-10were new homoisoflavanones, compound1was a new steroidal saponins and compound5was isolated from Ophiopogon genus for the first time.
2. A qualitative method was established by HPLC-MS" according to the MS" fragmentation patterns of6steroidal saponins. Steroidal saponins in O. japonicus and other formulas including O. japonicus could be identified rapidly without reference substance according to this method. It preliminary conquered that steroidal saponins were difficult to isolate and identify. Developed the fingerprint of O. japonicus by HPLC-MS and identified the peaks in fingerprint based on the HPLC-MS" method and literatures.
3. A quantitative method for simultaneous determination of three steroidal saponins was developed via HPLC-MS. It preliminary overcome the disadvantages that steroidal saponins were difficult to determine because of their low content and non chromophores in O. japonicus.The sample pretreatment of this method was simple compared with previous analytical methods. It could be used to distinguish different origin of O. japonicus and evaluated the samples from production process. It provided a fast and feabile way to improve the quality control level of O. japonicus.
4.4operating units of Shenmai Injection production process including12batches under normal operation condition (BNOC) and some batches under abnormal operation condition (BAOC) were prepared in lab scale. The HPLC-MS fingerprint combined with multivariate data analysis techniques were used to distinguish the abnormal compounds in fingerprint caused by the changes of operating parameters in BAOC. It could find the operating parameters affecting the quality of0. japonicus intermediates in the unit operation. This approach supplied a new strategy to evaluate the manufacture process of O. japonicus.
5. The performance of ceremic membranes and ultrafiltration membranes in the pass rate of three steroidal saponins and the clearance of total solid were investigated respectively and observed the best pore size of ceremic membrane and MWCO of ultrafiltration membrane. The suitable operating parameters were obtained by the research on the process of micro filtration and ultrafiltration. Preliminary studied the fouling problem of membrane material. This work not only provided the strategy for the ultrafiltration process of Shenmai Injection but also gave a new production process for O. japonicus. Researched on the loss of steroidal saponins in the activated carbon adsorption process and discussed its adsorption mechanism.
参麦注射液由红参和麦冬2味中药组成,是中药大品种。目前参麦注射液的质量控制仍存在不足之处—对组方药材浙麦冬的质量控制研究较少。鉴于此,本论文以浙麦冬为研究对象,开展其化学物质基础、质量分析方法和生产工艺的研究,希望通过本研究可以全面提高浙麦冬的质量控制水平,并为中药质量控制方法研究提供一个良好的范例。
本论文主要研究内容及学术贡献如下：
1.采用多种色谱技术系统研究浙麦冬的化学成分,分离得到12个化合物。综合运用质谱和核磁共振等波谱技术鉴定了11个化合物的结构,其中化合物8-10为新的高异黄酮类化合物,化合物1为新的甾体皂苷类化合物,化合物5为属内首次分离的甾体皂苷类化合物。
2.以6个甾体皂苷类化合物的多级质谱裂解规律为基础建立浙麦冬甾体皂苷类化学成分的HPLC-MSn定性方法。本方法可以在没有对照品的情况下快速辨识麦冬及其制剂中的甾体皂苷类化合物,初步解决了此类化合物分离难度大、结构鉴定困难的问题。建立浙麦冬药材的HPLC-MS指纹图谱,在HPLC-MSn定性分析方法的基础上并结合文献鉴定或推测了指纹图谱谱峰对应的化合物结构。
3.利用质谱的高灵敏度和高选择性建立同时测定浙麦冬3个甾体皂苷化合物的HPLC-MS定量分析方法,初步解决了麦冬甾体皂苷化合物由于含量低、紫外末端吸收,含量测定困难的问题。本方法相对于以往的分析方法具有样品预处理简单,节省分析时间等优势,可以用来区分不同产地的麦冬类药材和评价工艺中的麦冬中间体。本方法为提高浙麦冬质量控水平提供简单可行的途径。
4.实验室模拟浙麦冬4个操作单元的生产过程,每个操作单元设置12个正常批次和若干异常批次。利用HPLC-MS指纹图谱结合多变量数据分析技术辨识指纹图谱中与异常批次工艺参数变化相关的化合物,确立了单元操作中影响浙麦冬中间体质量的关键工艺参数。本研究为浙麦冬的工艺评价方法提供新的思路。
5.探索性研究了膜分离技术在浙麦冬生产中的应用,考察陶瓷膜和超滤膜对3个甾体皂苷通过率和总固含物清除率的影响,筛选出最佳孔径的陶瓷膜和最佳截留分子量的超滤膜以及合适的膜分离工艺参数,并初步研究了膜芯材料的污染问题。本研究既可以为参麦注射液的超滤工艺提供研究思路,又可以作为新工艺对参麦注射液工艺进行改进；研究活性炭吸附工艺过程中3个甾体皂苷的损失情况,探讨活性炭对3个甾体皂苷的吸附机理。
The quality control level of traditional Chinese medicine (TCM) industry is low and restricts its development. In order to improve the quality control level of TCM, the research on three critical units—hemical material basis, analytical methods and production processes in TCM quality control should be strengthed via modern science and technology.
Shenmai Injection is one of the major TCM composed of Panax ginseng (Hongshen) and Ophiopogon japonicus (Maidong). However, the quality control of Shenmai Injection is still inadequacies because the research on O. japonicus is less. In this thesis, chemical material basis, analytical methods and production processes for O. japonicus were studied. This study will raise the quality control level of O. japonicus and provide a good example for TCM quality control.
The main innovation and academic contributions of this dissertation are summarized as follows:
1. Column chromatography techniques were used to isolate the chemical constituents of O. japonicus grown in Zhejiang province.12compounds were obtained and11of them were identified via MS and NMR spectral analysis. Compounds8-10were new homoisoflavanones, compound1was a new steroidal saponins and compound5was isolated from Ophiopogon genus for the first time.
2. A qualitative method was established by HPLC-MS" according to the MS" fragmentation patterns of6steroidal saponins. Steroidal saponins in O. japonicus and other formulas including O. japonicus could be identified rapidly without reference substance according to this method. It preliminary conquered that steroidal saponins were difficult to isolate and identify. Developed the fingerprint of O. japonicus by HPLC-MS and identified the peaks in fingerprint based on the HPLC-MS" method and literatures.
3. A quantitative method for simultaneous determination of three steroidal saponins was developed via HPLC-MS. It preliminary overcome the disadvantages that steroidal saponins were difficult to determine because of their low content and non chromophores in O. japonicus.The sample pretreatment of this method was simple compared with previous analytical methods. It could be used to distinguish different origin of O. japonicus and evaluated the samples from production process. It provided a fast and feabile way to improve the quality control level of O. japonicus.
4.4operating units of Shenmai Injection production process including12batches under normal operation condition (BNOC) and some batches under abnormal operation condition (BAOC) were prepared in lab scale. The HPLC-MS fingerprint combined with multivariate data analysis techniques were used to distinguish the abnormal compounds in fingerprint caused by the changes of operating parameters in BAOC. It could find the operating parameters affecting the quality of0. japonicus intermediates in the unit operation. This approach supplied a new strategy to evaluate the manufacture process of O. japonicus.
5. The performance of ceremic membranes and ultrafiltration membranes in the pass rate of three steroidal saponins and the clearance of total solid were investigated respectively and observed the best pore size of ceremic membrane and MWCO of ultrafiltration membrane. The suitable operating parameters were obtained by the research on the process of micro filtration and ultrafiltration. Preliminary studied the fouling problem of membrane material. This work not only provided the strategy for the ultrafiltration process of Shenmai Injection but also gave a new production process for O. japonicus. Researched on the loss of steroidal saponins in the activated carbon adsorption process and discussed its adsorption mechanism.
引文
[1]张萍,杨燕,鄢丹,等.多指标成分含量测定与指纹图谱分析在中药制备工艺与质量控制中的应用.中华中医药杂志,2010,25(1)：120-123
[2]曹进,徐燕,王义明,等.中药指纹图谱获取方法验证及实际应用.世界科学技术-中医药现代化,2003,5(5)：31-38
[3]刘清飞,王义明,罗国安.多指标定量指纹图谱用于中药复方缓释制剂体外释放度的评价.中国中药杂志,2009,34(2)：143-147
[4]邱丽丽,容蓉,王薇.色谱及其联用技术在中药指纹图谱研究中的应用.山东中医药大学学报,2009,33(2)：165-168
[5]Wold S, Esbensen K, Geladi P. Principal component analysis. Chemometrics and intelligent laboratory systems,1987,2(1-3):37-52
[6]Geladi P, Kowalski B R. Partial least-squares regression-atutorial. Analytical Chimica Acta, 1986,185,1-17
[7]Kourti T. Process analytical technology beyond real-time analyzers:The role of multivariate analysis. Critical reviews in analytical chemistry,2006,36(3-4):257-278
[8]王北婴,王跃生,王焕魁.我国中药制药工业中亟需推广的高新技术.世界科学技术,2001,2(2)：18-24
[9]余伯阳,徐国钧.中药麦冬的资源利用研究.中草药,1995,26(4)：205-210
[10]程志红.无刺帽柱木及麦冬类药材活性部位的成分研究[博士学位论文]南京.中国药科大学,2003,27-48
[11]江洪波.中药涪麦冬的化学成分研究[硕士学位论文].成都.四川大学,2006,56-57
[12]Tang Y P, Yu B Y, Hu J, et al. Three New Homoisoflavanone Glycosides from the Bulbs of Ornithogalum caudatum. Journal of Natural Product,2002,65(2):218-220
[13]Matteo A, Miehelangelo P, Y X Zhu. Terpenoid glyeosides from Ophiopogon japonicus. Phytochemistry,1990,29(5):1696-1699
[14]陈纪军,朱祯禄,罗士德.慈溪麦冬苷A和B的结构.云南植物研究,2000,22(1)：97-102
[15]Duan, C L, Li, Y J, Li P, et al. Spirostanol Saponins from the Fibrous Roots of Ophiopogon japonicus (Thunb.) Ker-Gawl. Helvetica Chimica Acta,2010,93(2):227-232.
[16]张涛.中药麦冬中甾体皂苷类成分的研究[硕士学位论文].北京,军事医学科学院,2009.19-22
[17]Yoshiaki W, Shuichi S, Yoshiteru L, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners Ⅱ. Chemical and Pharmaceutical Bulletin,1983, 32(10):3986-3495
[18]李兴从,杨永平,李恒,等.大花沿阶草中甾体皂苷成分.云南植物研究,1990,12(S3)：1-3
[19]Yoshiaki W, Yasuaki H, Shuichi S, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners. (part V), Studies on the Constituents of subterranean part of Ophiopogon chekiangennis. K. Kimura et H Migo. Shoyakugaku Zasshi,1990,44(2): 117-121
[20]Yoshiaki W, Shuichi S, Akihiro T, et al. Studies on the Constituents of Ophiopogonis Tuber and Its congeners. Ⅳ. on the structure of Ophiopogonin A, B', C, C and'D. Chemical and Pharmaceutical Bulletin,1977,25(11):3049-3055
[21]肖培根.新编中药志.第1卷.北京.化学工业出版社,2001,481.-487
[22]Rgwat M. S. M., Negi D. S., Panwar. M.S., et al. Spermicidal potential and chemical analysis of Ophiopogonin intermedius(thizomes). Pharmazie,1988,43(5):143-145
[23]杨志,肖蓉,肖悼殷.川产麦冬化学成分的研究.华西药学杂志,1987,2(2)：57-60
[24]Wang J Z, Li M Y, Chen X B. A new C27-steroidal glycoside from Ophiopogon japonicus. Chinese Chemical Letters,2008,19(1):82-84
[25]Dai H F, Zhou J, Ding Z T, et al. Two new steroidal glyeosides from Ophiopogon Chinese Chemical Letters,2000,11(10):901-904
[26]Yoshiaki W, Shuichi S, Yoshiteru I, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners. Ⅲ Studies on the Constituents of subterranean part of ophiopogon ohwii Okuyama and O. jaburan (Kunth) LODD. Chemical and Pharmaceutical Bulletin,1984,32(10):3994-4002
[27]姜宇.麦冬须根的化学成分研究[硕士学位论文].沈阳,沈阳药科大学,2007,20-22
[28]余伯阳,徐国钧,平井康昭,等.湖北麦冬与短葶山麦冬的化学成分研究.中国药科大学学报,1988,3,209
[29]AkihiroT, Mashki K, Junzo S. Studies on the constituents of Ophiopogonis tuber. Ⅲ On the structure of Ophiopogonin D, Chemical and Pharmaceutical Bulletin,1973,21(2):308-311
[30]余伯阳,徐国钧,平井康昭,等.湖北麦冬与短葶山麦冬的化学成分研究Ⅱ.中国药科大学学报,1988,19,296
[31]Hisako K, Seiichi S, Akihiro T, et al. Studies on the constituents of Ophiopogonis Tuber Ⅰ. Isolation of steroidal glycosides from tuber of Ophiopogn japonics Ker-Gawler var.genuinus. Maxim.YakugakuZasshi,1968,88(6):710-714
[32]Zhang T, Zou P, Kang L P, et al. Two novel furostanol saponins from Ophiopogon japonicus. Journal of Asian Natural Products Research,2009,11(9):824-831
[33]Yu B Y, yasuiaki, Janzo S, et al. Comparative studies on the constituenis of Ophiopogonis tuber and its congeners, Ⅵ Studies on the Constituents of subterranean part of Liriope spicata var.prolifer and L. muscari (1). Chemical and Pharmaceutical Bulletin,1990,38(7):1931-1935
[34]Yu B Y, Qiu S X, kyaw Zaw, et al. Steroidal glycosides from the subterranean Parts of Liriope spicata var.prolifer. Phytochemistry,1996,43(1):201-206
[35]Yoshiaki W, Yasuaki H, Shuichi S, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners, (part V), Studies on the Constituents of subterranean part of Ophiopogon chekiangennis. K. Kimura et H Migo. Shoyakugaku Zasshi,1990,44(2): 117-121
[36]Akihiro T, Ryoji K, Amotsu S, et al. Studies on the constituents of Ophiopogonis tuber. Isolation of a novel class of Homoisoflavonoids and determination of their structure (1). Chemical and Pharmaceutical Bulletin,1980,28(5):1477-1487
[37]Norito K, Hiroyuki N, Tadayuki K, et al. Studies on the compounds of Ophiopogon Roots (China).YakugakuZasshi,1983,103(11):1133-1139
[38]Yoshiaki W, Shuichi S, Yoshiteru, et al Comparative studies on the constituents of ophiopogonis tuber and its congeners. IV. Studies on the Homoisoflavonoids of the O. jabura (Kunth) Lodd. Chemical and Pharmaceutical Bulletin,1985,33(12):5358-5363
[39]易进海.麦冬研究进展.华西药学杂志,1993,8(1)：32-38
[40]Hoang A.N., Van S.T, Porzel A., et al. Homoisoflavonoids from Ophiopogon japonicus Ker-Gawler. Phytochemistry,2003,62(7):1153-1158
[41]朱永新,严克东,屠国士.麦冬中高异黄酮的分离和鉴定.药学学报,1987,22(9)：679-684
[42]Akihiro T, Ryoji K, Amotsu S, et al. Studies on the constituents of Ophiopogonis TuberVl. Structures of Homoisoflavonoids.(2), Chemical and Pharmaceutical Bulletin,1980,28(7): 2039-2044
[43]Zhu Y X, Yan K D, Tu G S. Two homoisofiavones from Ophiopogon japonicus. Phytoehemistry,1987,26(10):2873-2874
[44]Cheng Z H, Wu T, S.W.Annie Bligh. cis-Eudesmane Sesquiterpene Glycosides from Liriope muscari and Ophiopogon japonicas. Journal of Natural Product,2004,67(10):1761-1763
[45]Duan C L, Kang Z Y, Lin C R. Two new homoisoflavonoids from the fibrous roots of Ophiopogon japonicus (Thunb.) Ker-Gawl. Journal of Asian Natural Products Research,2009, 11(10):876-879
[46]刘成基,曾诊,马蓓.麦冬化学成分的研究.中草药,1988,19(4)：10-11
[47]程志红,吴弢,余伯阳.短葶山麦冬化学成分的研究.中草药,2005,36(6)：823-826
[48]程志红,李林洲,刘楠,等.中药麦冬脂溶性化学成分的研究.中国药学杂志,2005,40(5)：337-341
[49]梅喆.麦冬须根的化学成分及含量测定研究[硕士学位论文].长春,吉林大学,2006,35-42
[50]吴笑如,徐德生,冯怡,等HPLC-ELSD测定川麦冬须根大孔树脂富集物中麦冬皂苷D、D’的含量.中成药,2006,28(11)：1638-1640
[51]吴涛,程志红,徐珞珊,等HPLC-ELSD法测定湖北麦冬中主要皂苷的含量.中草药,2000,31(3)：175-177
[52]唐晓清,余伯阳,徐德然,等HPLC-ELSD法测定麦冬中甾体皂甙元的含量.中国药科大学学报,2001,32(4)：270-272
[53]Wu L. On-line determination of effect of Ophiopogon japonicus on scavenging superoxide anion radicals by HPLC-DAD coupled with chemiluminescence. Zhongguo Yaoke Daxue Xuebao,2010,41(2):141-145
[54]王志斌,陈有根,李慧HPLC-ELSD法测定麦冬药材及其提取物中果糖的含量.中国中医药信息杂志,2007,14(2)：47-48
[55]胥秀英,郑一敏,傅善权,等.麦冬须根与麦冬块根中麦冬皂苷Ra和慈溪麦冬皂苷A含量比较.中国中药杂志,2009,19(34)：2532-2533
[56]黄宝美,姚程炜,莫金垣,等.高效毛细管电泳安培法测定麦冬中薯蓣皂甙元的含量.中山大学学报(自然科学版),2010,49(2)：65-71
[57]王红英,钱斯日古楞,赵前程,等.3,5-二硝基水杨酸比色法测定麦冬多糖含量.沈阳农业大学学报,2005.36(5)：628-630
[58]Ye G G, Ye M, Guo D A, et al. Determination of Homoisoflavonoids in Ophiopogon japonicus by RP-HPLC. Chromatographia,2005,61(3):121-125
[59]王远,秦民坚,戚近,等.近红外漫反射光谱法测定麦冬的多糖含量.光谱学与光谱分析,2009,29(10)：2677-2680
[60]马军守,别继明,金虹.川麦冬及其须根中多糖和黄酮类有效成分定量分析.西南科技大学学报,2008,23(1)：83-86
[61]林以宁,余伯阳.化学发光法测定麦冬类药材中总酚性成分含量.林以宁,余伯阳.化学发光法测定麦冬类药材中总酚性成分含量.中药材,2006,29(8)：797-799
[62]Ye M, Guo D A, Ye G G. Analysis of Homoisoflavonoids in Ophiopogon japonicus by HPLC-DAD-ESI-MS". Journal of American Society Mass for Spectrometry,2005,16(2): 234-243
[63]沈宏林,向能军,许永高,等GC-MS分析麦冬中脂溶性成分.光谱实验室，25(4)：669-672
[64]谭小燕,罗乔奇,马郑红,等.不同产地麦冬1H-NMR模式识别研究.中草药,2009,40(5)：793-797
[65]Liu L, Lu Y, Shao Q, et al. Binary chromatographic fingerprinting for quality evaluation of Radix Ophiopogonis by high performance liquid chromatography coupled with ultraviolet and evaporative light-scattering detectors. Journal of Separation Science,2007,30(16):2628-2637
[66]董政起,朱静,金光株.麦冬与山麦冬的HPLC指纹图谱比较研究.特产研究,2007,29(1)：20--22
[67]韩凤梅,李紫,蔡敏,等.山麦冬的HPLC/ESI-MS特征指纹图谱研究.中草药,2005,36(9)：1395-1397
[68]林以宁,志田保夫,袁博,等.不同产地麦冬的指纹图谱比较研究.中国药科大学学报,2005,36(6)：538-542
[69]吴永江,崔勤敏,程翼宇.制备工艺对参麦注射液质量的影响.中国中药杂志,2005,30(9)：662-664
[70]钟永红,周南,钱琳.参麦注射液制备工艺研究.中医药导报,2007,13(6)：106-107
[71]崔勤敏.参麦注射液指纹图谱分析与工艺研究.[硕士学位论文].杭州,浙江大学,2005,49-55
[72]胡楚楚.参麦注射液质量评价方法研究.[硕士学位论文].杭州,浙江大学,2006,12-27
[73]张全英,施爱明,袁心慧.参麦注射液中人参皂苷含量测定.中草药,2000,31(10)：750-751
[74]陈闽军,吴永江,程翼宇.高效液相色谱法用于参麦注射液中人参皂苷的含量测定与批次间一致性考察.中国药学杂志.2003,38(8)：623-625.
[75]王志超,潘灏白,刘卫红.超滤法和水醇法制备参麦注射液的实验比较研究.中国药房,1994,5(1)：11-12
[76]Eddy, C. R., Wall, M. E.,& Scott M. K., Catalog of infrared absorption spectra of steroidal sapogenin aetates. Analytical Chemistry,1953,25(2):266-271
[77]天然药物化学.沈阳.吴立军主编.第五版.89-91
[78]Agrawal, P. K, Jain, D. C, Pathak, A. K. et al. NMR Spectroscopy of Steroidal Sapogenins and Steroidal Saponins:an Update. Magnetic Resonance in Chemistry,1995,33(12):923-953
[79]Kuroda M, Mimaki Y, Ori K, et al. Steroidal Glycosides from the Bulbs of Ornithogalum thyrsoides. Journal of Natural Products,2004,67(10):1690-1696
[80]Namikoshi M, Saitoh T. Homoisoflavonoids and Related Compounds. IV. Absolute Configurations of Homoisoflavonoids from Caesalpinia sappan L. Chemical and Pharmaceutical Bulletin.1987,35(9):3597-3602.
[81]Domon B, Costello C. A Systematic Nomenclature for Carbohydrate Fragmentations in FAB-MS/MS Spectra of Glycoconjμgates. Journal Glycoconjugates,1988,5(4):397-409
[82]董洁,郭立玮,陈丹丹,等.0.2μm无机陶瓷膜微滤对黄芩等7种中药主要指标性成分转移率的影响.南京中医药大学学报,2003,19(3)：148-150
[83]陈丹丹,郭立玮,刘爱国,等.0.2μm无机陶瓷膜微滤对生地黄、黄芪水提液物理化学参数影响的初步研究.南京中医药大学学报,2002,18(3)：153-155
[84]潘永兰,郭立玮,黄耀洲,等.pH对无机陶瓷膜微滤中药水提液膜过程的影响.世界科技研究与发展,2008,30(6)：709-710
[85]王天瑶,潘永兰,郭立玮.金银花等中药水提取液溶液环境与陶瓷膜微滤过程稳定通量、阻力分布相关性的研究.时珍国医国药,2008,19(5)：2571-2573
[86]刘陶世,郭立玮,袁铸人,等.无机陶瓷膜微滤技术精制7种根及根茎类中药,水提液的研究.中成药,2001,23(5)：473-476
[87]于庆立,贾丽萍,李英刚,等.用膜分离法提取清肝颗粒的工艺研究.中医药学报,2008,36(2)：51-52
[88]尹楠,李红阳,彭国平,等.超滤法去除中药注射液中的细菌内毒素.中国医药工业杂志,2008,39(12)：927-929
[89]张学著,张萍菊.超滤法提高通宣理肺口服液质量的研究.中国医药工业杂志,2002,33(6)：283-285
[90]彭国平,郭立玮,徐丽华,等.超滤技术应用对中药成分的影响.南京中医药大学学报(自然科学版)：2002,18(6)：339-341
[91]罗吉,钟美好,肖凯军,等.多级膜浓缩鼻炎康提取液的研究.中国中药杂志,2008,33(17)：2108-2112
[92]王丹,龚涛,孙冰,等.胶团超滤法制备丹参注射液.中成药,2003,25(6)：441-443
[93]袁亮,周显宏,肖凯军,等.多级膜浓缩黄芩苷提取液的研究.现代食品科技,2008,24(3)：237-240
[94]叶勇,张永波.复方中药膜分离工艺和节能性研究.时珍国医国药,2008,19(8)：1884-1885
[95]Sim L, Shu L, Jegatheesan V, et al. Effect of operating parameters and cleaning on the performance of ceramic membranes treating partially clarified sugar cane juice. Separation Science and Technology,2009,44(12):3506-3537
[96]Sun Y, Li A M, Zhang Q X, et al. Adsorptive Separation of Tannic Acid from Aqueous Solution by Polymeric Resins. Separation Science and Technology,2008,43(2):389-402
[97]Valderrama C, Cortina J L, Farran A, et al. Kinetics of sorption of polyaromatic hydrocarbo ns onto granular activated carbon and Macronet hyper-cross-linked polymers (MN200). Journal of Colloid and Interface Science,2007,310(1):35-44
[2]曹进,徐燕,王义明,等.中药指纹图谱获取方法验证及实际应用.世界科学技术-中医药现代化,2003,5(5)：31-38
[3]刘清飞,王义明,罗国安.多指标定量指纹图谱用于中药复方缓释制剂体外释放度的评价.中国中药杂志,2009,34(2)：143-147
[4]邱丽丽,容蓉,王薇.色谱及其联用技术在中药指纹图谱研究中的应用.山东中医药大学学报,2009,33(2)：165-168
[5]Wold S, Esbensen K, Geladi P. Principal component analysis. Chemometrics and intelligent laboratory systems,1987,2(1-3):37-52
[6]Geladi P, Kowalski B R. Partial least-squares regression-atutorial. Analytical Chimica Acta, 1986,185,1-17
[7]Kourti T. Process analytical technology beyond real-time analyzers:The role of multivariate analysis. Critical reviews in analytical chemistry,2006,36(3-4):257-278
[8]王北婴,王跃生,王焕魁.我国中药制药工业中亟需推广的高新技术.世界科学技术,2001,2(2)：18-24
[9]余伯阳,徐国钧.中药麦冬的资源利用研究.中草药,1995,26(4)：205-210
[10]程志红.无刺帽柱木及麦冬类药材活性部位的成分研究[博士学位论文]南京.中国药科大学,2003,27-48
[11]江洪波.中药涪麦冬的化学成分研究[硕士学位论文].成都.四川大学,2006,56-57
[12]Tang Y P, Yu B Y, Hu J, et al. Three New Homoisoflavanone Glycosides from the Bulbs of Ornithogalum caudatum. Journal of Natural Product,2002,65(2):218-220
[13]Matteo A, Miehelangelo P, Y X Zhu. Terpenoid glyeosides from Ophiopogon japonicus. Phytochemistry,1990,29(5):1696-1699
[14]陈纪军,朱祯禄,罗士德.慈溪麦冬苷A和B的结构.云南植物研究,2000,22(1)：97-102
[15]Duan, C L, Li, Y J, Li P, et al. Spirostanol Saponins from the Fibrous Roots of Ophiopogon japonicus (Thunb.) Ker-Gawl. Helvetica Chimica Acta,2010,93(2):227-232.
[16]张涛.中药麦冬中甾体皂苷类成分的研究[硕士学位论文].北京,军事医学科学院,2009.19-22
[17]Yoshiaki W, Shuichi S, Yoshiteru L, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners Ⅱ. Chemical and Pharmaceutical Bulletin,1983, 32(10):3986-3495
[18]李兴从,杨永平,李恒,等.大花沿阶草中甾体皂苷成分.云南植物研究,1990,12(S3)：1-3
[19]Yoshiaki W, Yasuaki H, Shuichi S, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners. (part V), Studies on the Constituents of subterranean part of Ophiopogon chekiangennis. K. Kimura et H Migo. Shoyakugaku Zasshi,1990,44(2): 117-121
[20]Yoshiaki W, Shuichi S, Akihiro T, et al. Studies on the Constituents of Ophiopogonis Tuber and Its congeners. Ⅳ. on the structure of Ophiopogonin A, B', C, C and'D. Chemical and Pharmaceutical Bulletin,1977,25(11):3049-3055
[21]肖培根.新编中药志.第1卷.北京.化学工业出版社,2001,481.-487
[22]Rgwat M. S. M., Negi D. S., Panwar. M.S., et al. Spermicidal potential and chemical analysis of Ophiopogonin intermedius(thizomes). Pharmazie,1988,43(5):143-145
[23]杨志,肖蓉,肖悼殷.川产麦冬化学成分的研究.华西药学杂志,1987,2(2)：57-60
[24]Wang J Z, Li M Y, Chen X B. A new C27-steroidal glycoside from Ophiopogon japonicus. Chinese Chemical Letters,2008,19(1):82-84
[25]Dai H F, Zhou J, Ding Z T, et al. Two new steroidal glyeosides from Ophiopogon Chinese Chemical Letters,2000,11(10):901-904
[26]Yoshiaki W, Shuichi S, Yoshiteru I, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners. Ⅲ Studies on the Constituents of subterranean part of ophiopogon ohwii Okuyama and O. jaburan (Kunth) LODD. Chemical and Pharmaceutical Bulletin,1984,32(10):3994-4002
[27]姜宇.麦冬须根的化学成分研究[硕士学位论文].沈阳,沈阳药科大学,2007,20-22
[28]余伯阳,徐国钧,平井康昭,等.湖北麦冬与短葶山麦冬的化学成分研究.中国药科大学学报,1988,3,209
[29]AkihiroT, Mashki K, Junzo S. Studies on the constituents of Ophiopogonis tuber. Ⅲ On the structure of Ophiopogonin D, Chemical and Pharmaceutical Bulletin,1973,21(2):308-311
[30]余伯阳,徐国钧,平井康昭,等.湖北麦冬与短葶山麦冬的化学成分研究Ⅱ.中国药科大学学报,1988,19,296
[31]Hisako K, Seiichi S, Akihiro T, et al. Studies on the constituents of Ophiopogonis Tuber Ⅰ. Isolation of steroidal glycosides from tuber of Ophiopogn japonics Ker-Gawler var.genuinus. Maxim.YakugakuZasshi,1968,88(6):710-714
[32]Zhang T, Zou P, Kang L P, et al. Two novel furostanol saponins from Ophiopogon japonicus. Journal of Asian Natural Products Research,2009,11(9):824-831
[33]Yu B Y, yasuiaki, Janzo S, et al. Comparative studies on the constituenis of Ophiopogonis tuber and its congeners, Ⅵ Studies on the Constituents of subterranean part of Liriope spicata var.prolifer and L. muscari (1). Chemical and Pharmaceutical Bulletin,1990,38(7):1931-1935
[34]Yu B Y, Qiu S X, kyaw Zaw, et al. Steroidal glycosides from the subterranean Parts of Liriope spicata var.prolifer. Phytochemistry,1996,43(1):201-206
[35]Yoshiaki W, Yasuaki H, Shuichi S, et al. Comparative Studies on the Constituents of Ophiopogonis Tuber and Its congeners, (part V), Studies on the Constituents of subterranean part of Ophiopogon chekiangennis. K. Kimura et H Migo. Shoyakugaku Zasshi,1990,44(2): 117-121
[36]Akihiro T, Ryoji K, Amotsu S, et al. Studies on the constituents of Ophiopogonis tuber. Isolation of a novel class of Homoisoflavonoids and determination of their structure (1). Chemical and Pharmaceutical Bulletin,1980,28(5):1477-1487
[37]Norito K, Hiroyuki N, Tadayuki K, et al. Studies on the compounds of Ophiopogon Roots (China).YakugakuZasshi,1983,103(11):1133-1139
[38]Yoshiaki W, Shuichi S, Yoshiteru, et al Comparative studies on the constituents of ophiopogonis tuber and its congeners. IV. Studies on the Homoisoflavonoids of the O. jabura (Kunth) Lodd. Chemical and Pharmaceutical Bulletin,1985,33(12):5358-5363
[39]易进海.麦冬研究进展.华西药学杂志,1993,8(1)：32-38
[40]Hoang A.N., Van S.T, Porzel A., et al. Homoisoflavonoids from Ophiopogon japonicus Ker-Gawler. Phytochemistry,2003,62(7):1153-1158
[41]朱永新,严克东,屠国士.麦冬中高异黄酮的分离和鉴定.药学学报,1987,22(9)：679-684
[42]Akihiro T, Ryoji K, Amotsu S, et al. Studies on the constituents of Ophiopogonis TuberVl. Structures of Homoisoflavonoids.(2), Chemical and Pharmaceutical Bulletin,1980,28(7): 2039-2044
[43]Zhu Y X, Yan K D, Tu G S. Two homoisofiavones from Ophiopogon japonicus. Phytoehemistry,1987,26(10):2873-2874
[44]Cheng Z H, Wu T, S.W.Annie Bligh. cis-Eudesmane Sesquiterpene Glycosides from Liriope muscari and Ophiopogon japonicas. Journal of Natural Product,2004,67(10):1761-1763
[45]Duan C L, Kang Z Y, Lin C R. Two new homoisoflavonoids from the fibrous roots of Ophiopogon japonicus (Thunb.) Ker-Gawl. Journal of Asian Natural Products Research,2009, 11(10):876-879
[46]刘成基,曾诊,马蓓.麦冬化学成分的研究.中草药,1988,19(4)：10-11
[47]程志红,吴弢,余伯阳.短葶山麦冬化学成分的研究.中草药,2005,36(6)：823-826
[48]程志红,李林洲,刘楠,等.中药麦冬脂溶性化学成分的研究.中国药学杂志,2005,40(5)：337-341
[49]梅喆.麦冬须根的化学成分及含量测定研究[硕士学位论文].长春,吉林大学,2006,35-42
[50]吴笑如,徐德生,冯怡,等HPLC-ELSD测定川麦冬须根大孔树脂富集物中麦冬皂苷D、D’的含量.中成药,2006,28(11)：1638-1640
[51]吴涛,程志红,徐珞珊,等HPLC-ELSD法测定湖北麦冬中主要皂苷的含量.中草药,2000,31(3)：175-177
[52]唐晓清,余伯阳,徐德然,等HPLC-ELSD法测定麦冬中甾体皂甙元的含量.中国药科大学学报,2001,32(4)：270-272
[53]Wu L. On-line determination of effect of Ophiopogon japonicus on scavenging superoxide anion radicals by HPLC-DAD coupled with chemiluminescence. Zhongguo Yaoke Daxue Xuebao,2010,41(2):141-145
[54]王志斌,陈有根,李慧HPLC-ELSD法测定麦冬药材及其提取物中果糖的含量.中国中医药信息杂志,2007,14(2)：47-48
[55]胥秀英,郑一敏,傅善权,等.麦冬须根与麦冬块根中麦冬皂苷Ra和慈溪麦冬皂苷A含量比较.中国中药杂志,2009,19(34)：2532-2533
[56]黄宝美,姚程炜,莫金垣,等.高效毛细管电泳安培法测定麦冬中薯蓣皂甙元的含量.中山大学学报(自然科学版),2010,49(2)：65-71
[57]王红英,钱斯日古楞,赵前程,等.3,5-二硝基水杨酸比色法测定麦冬多糖含量.沈阳农业大学学报,2005.36(5)：628-630
[58]Ye G G, Ye M, Guo D A, et al. Determination of Homoisoflavonoids in Ophiopogon japonicus by RP-HPLC. Chromatographia,2005,61(3):121-125
[59]王远,秦民坚,戚近,等.近红外漫反射光谱法测定麦冬的多糖含量.光谱学与光谱分析,2009,29(10)：2677-2680
[60]马军守,别继明,金虹.川麦冬及其须根中多糖和黄酮类有效成分定量分析.西南科技大学学报,2008,23(1)：83-86
[61]林以宁,余伯阳.化学发光法测定麦冬类药材中总酚性成分含量.林以宁,余伯阳.化学发光法测定麦冬类药材中总酚性成分含量.中药材,2006,29(8)：797-799
[62]Ye M, Guo D A, Ye G G. Analysis of Homoisoflavonoids in Ophiopogon japonicus by HPLC-DAD-ESI-MS". Journal of American Society Mass for Spectrometry,2005,16(2): 234-243
[63]沈宏林,向能军,许永高,等GC-MS分析麦冬中脂溶性成分.光谱实验室，25(4)：669-672
[64]谭小燕,罗乔奇,马郑红,等.不同产地麦冬1H-NMR模式识别研究.中草药,2009,40(5)：793-797
[65]Liu L, Lu Y, Shao Q, et al. Binary chromatographic fingerprinting for quality evaluation of Radix Ophiopogonis by high performance liquid chromatography coupled with ultraviolet and evaporative light-scattering detectors. Journal of Separation Science,2007,30(16):2628-2637
[66]董政起,朱静,金光株.麦冬与山麦冬的HPLC指纹图谱比较研究.特产研究,2007,29(1)：20--22
[67]韩凤梅,李紫,蔡敏,等.山麦冬的HPLC/ESI-MS特征指纹图谱研究.中草药,2005,36(9)：1395-1397
[68]林以宁,志田保夫,袁博,等.不同产地麦冬的指纹图谱比较研究.中国药科大学学报,2005,36(6)：538-542
[69]吴永江,崔勤敏,程翼宇.制备工艺对参麦注射液质量的影响.中国中药杂志,2005,30(9)：662-664
[70]钟永红,周南,钱琳.参麦注射液制备工艺研究.中医药导报,2007,13(6)：106-107
[71]崔勤敏.参麦注射液指纹图谱分析与工艺研究.[硕士学位论文].杭州,浙江大学,2005,49-55
[72]胡楚楚.参麦注射液质量评价方法研究.[硕士学位论文].杭州,浙江大学,2006,12-27
[73]张全英,施爱明,袁心慧.参麦注射液中人参皂苷含量测定.中草药,2000,31(10)：750-751
[74]陈闽军,吴永江,程翼宇.高效液相色谱法用于参麦注射液中人参皂苷的含量测定与批次间一致性考察.中国药学杂志.2003,38(8)：623-625.
[75]王志超,潘灏白,刘卫红.超滤法和水醇法制备参麦注射液的实验比较研究.中国药房,1994,5(1)：11-12
[76]Eddy, C. R., Wall, M. E.,& Scott M. K., Catalog of infrared absorption spectra of steroidal sapogenin aetates. Analytical Chemistry,1953,25(2):266-271
[77]天然药物化学.沈阳.吴立军主编.第五版.89-91
[78]Agrawal, P. K, Jain, D. C, Pathak, A. K. et al. NMR Spectroscopy of Steroidal Sapogenins and Steroidal Saponins:an Update. Magnetic Resonance in Chemistry,1995,33(12):923-953
[79]Kuroda M, Mimaki Y, Ori K, et al. Steroidal Glycosides from the Bulbs of Ornithogalum thyrsoides. Journal of Natural Products,2004,67(10):1690-1696
[80]Namikoshi M, Saitoh T. Homoisoflavonoids and Related Compounds. IV. Absolute Configurations of Homoisoflavonoids from Caesalpinia sappan L. Chemical and Pharmaceutical Bulletin.1987,35(9):3597-3602.
[81]Domon B, Costello C. A Systematic Nomenclature for Carbohydrate Fragmentations in FAB-MS/MS Spectra of Glycoconjμgates. Journal Glycoconjugates,1988,5(4):397-409
[82]董洁,郭立玮,陈丹丹,等.0.2μm无机陶瓷膜微滤对黄芩等7种中药主要指标性成分转移率的影响.南京中医药大学学报,2003,19(3)：148-150
[83]陈丹丹,郭立玮,刘爱国,等.0.2μm无机陶瓷膜微滤对生地黄、黄芪水提液物理化学参数影响的初步研究.南京中医药大学学报,2002,18(3)：153-155
[84]潘永兰,郭立玮,黄耀洲,等.pH对无机陶瓷膜微滤中药水提液膜过程的影响.世界科技研究与发展,2008,30(6)：709-710
[85]王天瑶,潘永兰,郭立玮.金银花等中药水提取液溶液环境与陶瓷膜微滤过程稳定通量、阻力分布相关性的研究.时珍国医国药,2008,19(5)：2571-2573
[86]刘陶世,郭立玮,袁铸人,等.无机陶瓷膜微滤技术精制7种根及根茎类中药,水提液的研究.中成药,2001,23(5)：473-476
[87]于庆立,贾丽萍,李英刚,等.用膜分离法提取清肝颗粒的工艺研究.中医药学报,2008,36(2)：51-52
[88]尹楠,李红阳,彭国平,等.超滤法去除中药注射液中的细菌内毒素.中国医药工业杂志,2008,39(12)：927-929
[89]张学著,张萍菊.超滤法提高通宣理肺口服液质量的研究.中国医药工业杂志,2002,33(6)：283-285
[90]彭国平,郭立玮,徐丽华,等.超滤技术应用对中药成分的影响.南京中医药大学学报(自然科学版)：2002,18(6)：339-341
[91]罗吉,钟美好,肖凯军,等.多级膜浓缩鼻炎康提取液的研究.中国中药杂志,2008,33(17)：2108-2112
[92]王丹,龚涛,孙冰,等.胶团超滤法制备丹参注射液.中成药,2003,25(6)：441-443
[93]袁亮,周显宏,肖凯军,等.多级膜浓缩黄芩苷提取液的研究.现代食品科技,2008,24(3)：237-240
[94]叶勇,张永波.复方中药膜分离工艺和节能性研究.时珍国医国药,2008,19(8)：1884-1885
[95]Sim L, Shu L, Jegatheesan V, et al. Effect of operating parameters and cleaning on the performance of ceramic membranes treating partially clarified sugar cane juice. Separation Science and Technology,2009,44(12):3506-3537
[96]Sun Y, Li A M, Zhang Q X, et al. Adsorptive Separation of Tannic Acid from Aqueous Solution by Polymeric Resins. Separation Science and Technology,2008,43(2):389-402
[97]Valderrama C, Cortina J L, Farran A, et al. Kinetics of sorption of polyaromatic hydrocarbo ns onto granular activated carbon and Macronet hyper-cross-linked polymers (MN200). Journal of Colloid and Interface Science,2007,310(1):35-44