摘要
搏心通的原药材葱白为百合科植物分葱Allium fisturosum L. var. caespitosum Makio的干燥鳞茎,性辛温,功效发表、通阳、解毒,可治疗多种疾病。
采用超临界CO2萃取技术(SFE )提取搏心通原料药,并对原料药运用气质联用技术进行化学成分分析,检索出了八类成分:有机硫化物、有机胺、杂环化合物、醛、酮、酯、脂肪酸(酯)、长链烷烃。我们还与传统的水蒸气蒸馏法(SDE)进行了比较研究。通过查阅文献资料和药理实验,证明其中的有机硫化物和脂肪酸两类成分为搏心通的有效成分,并针对这两类成分的结构进行详细的分析。
本实验将脂肪酸衍生化成相应的酯(甲基酯)解决了脂肪酸GC中拖尾的问题,然后进行GC-MS检索,分离并鉴定出9种脂肪酸(甲酯),计算出它们的相对百分含量。含量较高的依次是亚油酸甲酯(C19H34O2) 62.44%、棕榈酸甲酯(C17H34O2) 21.34%、亚麻酸甲酯(C19H3202) 9.65%。在对有机硫化物的分析中,本实验从生源途径入手,针对硫化物形成过程,来验证检索结构的准确程度。
由于搏心通软胶囊原料药为油脂类成分,本实验按照药典(附录IX N)上规定对其理化常数进行了测定,从而确定酸值、皂化值、羟值、碘值的有效范围应分别控制在为40~43、177~180、127~130、40~43。
实验采用气相色谱法对脂肪酸类成分的质量进行研究。首先确立最佳的色谱条件,色谱柱为:HP-INNOWAX(30 m×0.53 mm×1 um),程序升温法:120℃保持2 min,以10℃/min的速度升到180℃,保持2 min,以2℃/min速度升到184℃,以1℃/min速度升到190℃;然后采用甲酯化法对样品进行衍生化处理,方法学考察结果表明精密度良好,加样回收率为97%~103%。证明该方法操作简便,结果准确可靠。
运用元素分析仪测定药物总硫,方法学考察结果表明:精密度良好,加样回收率为99%~104%,证明了该方法准确可行。结合化学成分分析的结果,将硫元素全部换算成含量最高的2,5-二甲基噻吩的含量,为8.6%。
The original medicines of boxintong come from Allium fisturosum L. var. caespitosum Makio which is a plant belonging to family of lily, genera of shallot. It has been used as folk medicine to treat many diseases.
The chemical compositions of raw materials of boxintong (SFE) have been studied by GC-MS method. Eight kinds of constituents were got and identified, which are organic sulfide compounds, organic amine, heterocycles, aldehyde, ketone, ester, fatty acid (ester), and long-chain alkanes, compairing with traditional method steam distillation extraction(SDE). Organic sulfide compounds and fatty acid have a lot of effect on pharmacological aspects and extensive curative effects by researchs and reports.
Nine fatty acid compounds were also got and identified by GC-MS. The total content of the fatty acids was 7%. The content of linoleic acid was the highest (62.44%)and palmitic acid was the next (21.34%). In the research of organic sulfide compounds, we analysis their structures by biogenesis pathway.
As fatty oil, we determine physicochemical constant of boxintong with method from the Chinese Pharmacopoeia. So that we could provisions the effective ranges of the raw materials: acid value 40~43, saponification value 177~180,hydroxyl value 127~130, iodine value 40~43.
A capillary gas chromatographic (GC) method equipped with flame ionization detection was developed for the simultaneous analysis of linoleic acid in boxintong. Compounds were separated on an HP-INNOWAX(30 m×0.53 mm×1 um). The polar column was 120℃for 2min programmed to 180℃at 10℃/min, 180℃for 2 min, programmed to 184℃at 2℃/min, 184℃for 1 min, programmed to 190℃at 1℃/min.;detector temperature 250℃. The orthogonal design was adopted to optimize the derivation reaction conditions of fatty oil. The calibration curves were linear in the range of 0.107~0.321 ug (R2=0.9991) . The average recovery of linoleic acid was 97% ~103% (RSD=1.3%). The assay method was simple, rapid, reliable and reproducible and provided a quantitative basis for the quality assessment of boxintong.
Determine the total sulfide element of drugs by elemental Analyzers at the first time. The result shows that its precision was under 3%, and the recoveries ranges were of 99%~104%. The proposed method is rapid and suitable for the determination of total sulfide element. Then all sulfide elements will be converted to organic sulfide 2,5 - dimethyl thiophene the highest content. It is 8.6%.
引文
[1].李为民,金波,冯毅凡.中药现代化与超临界流体萃取技术[M].北京:中国医药科技出版社,2002:117.
[2].郑希林,李瑞华.国内用超临界萃取技术研究中药近况[J].时珍国医国药,2000,1(8)
[3].陈新,崔健,张英华.超临界流体萃取法在提取天然植物中有效成分的应用[J].长春中医学院学报,1997,13(9):55.
[4].袁珂,殷明文.气相色谱-质谱法分析含羞草挥发油的化学成分[J].质谱学报,2006,27(1):50-52.
[5].凌育赵,曾满枝,陈祖佑.严志云沙姜挥发油化学成分的GC/MS分析[J].河南工业大学学报(自然科学版),2006,27(1):75-77.
[6]. Christine Robles, Suzanne Garzino. Essential Oil Composition of Cistus Albidus Leaves[J].Phytochemistry,1998,48(8):1341-1345.
[7]. Stella Kokkini, Regina Karousou, et al. Autumn essential oil of greek oregano[J]. Phytochemistry,1997,44(5):883-886.
[8]. B.R. Rajeswara Raoa, P.N. Kaul , K.V. Syamasundar , S. Rameshb. Chemical profiles of primary and secondary essential oils of palmarosa (Cymbopogon martinii (Roxb.)Wats var. motia Burk.)[J]. Industrial Crops and Products,2005,21:121–127.
[9]. Noel G. Porter , Alistair L. Wilkins. Chemical, physical and antimicrobial properties of essential oils of Leptospermum scoparium and Kunzea ericoides[J]. Phytochemistry, 1998,50: 407-415.
[10]. Philip J. Marriotta , Robert Shelliea, Charles Cornwell. Gas chromatographic technologies for the analysis of essential oils[J]. Journal of Chromatography A, 2001,936: 1–22.
[11]. Vincenzo De Feoa, Francesco De Simonea, Felice Senatore. Potential allelochemicals from the essential oil of Ruta graveolens[J]. Phytochemistry,2002,61: 573–578.
[12].邹忠梅,于德泉,丛浦珠.葱属植物化学及药理研究进展[J].药学学报,1999,34(5):395-400.
[13].吴雁,彭军鹏,姚利强,等.葱属植物挥发油研究[J].沈陌芍学院学报,1993,10(1):45-47.
[14].彭军鹏,乔艳秋,肖克岳,等.葱属植物挥发油研究Ⅲ薤(Allium chinense G.Don)挥发油成分的研究.中国药物化学杂志,1994,4(4):282-284.
[15].娄红祥,左春旭,李铣.大蒜化学成分的研究进展[M].国外医药·植物药分册,1991,6(2):55-59.
[16].陈能熠,伍睿,陈丽,等.大蒜研究进展[J].天然产物研究与开发,2002,12(2):67-74.
[17].张卿,高尔.薤白的研究进展[J].中国中药杂志,2003,28(2):105-107.
[18].姜永,王乃利,姚新生.中药薤自的研究进展[J].中国中药杂志,1999,12(5):74-79.
[19]. Nyunt Phay, Takako Higashiyama, Masahisa Tsuji, et al. An antifungal compound from roots of Welsh onion[J]. Phytochemistry ,1999,52: 271-274.
[20]. May-Chien Kuo, Chi-Tang Ho. Volatile Constituents of the Distilled Oils of Welsh Onions (Alliumfistulosum L. Variety Maichuon) and Scallions (Allium fistulosum L.Variety Caespitosum)[J]. J. Agric. FoodChem,1992,40:111-117.
[21].田莉,杨秀伟,陶海燕.大蒜化学成分的气一质联用分析[J].天然产物研究与开发,2005,17(5):533-538.
[22].王建,贾斌.大蒜油中大蒜素等含硫化合物的气相色谱一质谱法测定[J].中国饲料,2001,23:29.
[23].刘广军.GC/MS分析大蒜中的挥发油[J].聊城大学学报(自然科学版),2004,17(3):39-42.
[24].李静,唐纪琳,尹建元,等.气质联机法测定西洋参花蕾及人参花蕾中脂肪酸成分[J].人参研究,1999,11(2):31-32.
[25].马爱华,周晓东,乐巍.10种果实类中药中脂肪酸的分离鉴定和含量测定[J].时珍国药研究,1998,9(1):53.
[26]. Ghita Studsgaard Nielsen, Lone Melchior Larsen, Leif Poll. Formation of Volatile Compounds in Model Experiments with Crude Leek (Allium ampeloprasum Var. Lancelot) Enzyme Extract and Linoleic Acid or Linolenic Acid[J]. J. Agric. Food Chem.,2004, 52:2315-2321.
[27].林文群,阵忠.白花益母草种子的化学成分分析[J].天然产物研究,2001,13(4):40-43.
[28].付松,徐先顺,向奋飞.保健油脂中多不饱和脂肪酸的GC一MS分析[J].分析测试学报,2006,24:339-342.
[29].葛发欢,史庆龙,沈永康,等.菜籽油脚中天然VE脂肪油的超临界CO2萃取分离及其GC-MS分析[J].中药材,1997,20(4):189-191.
[30].陈炳华,刘剑秋,林文群,等.海边月见草种子油中脂肪酸组成的分析[J].福建师范太学学报(自然科学版),2001,17(1):75-78.
[31].李嵘,张媛,王喆之.苹果种籽中脂肪酸组分分析[J].植物生理学通讯,2006,42(5):955-957.
[32].徐水祥,周淡宜,徐飞鹤,等.甲鱼油中多不饱和脂肪酸的提取与纯化研究[J].浙江省医学科学院学报,2005,61:44-46.
[33].林秀英,刘传华,吕承坤,等.生、炒益智仁脂肪的气相色谱测定[J].山东医药工业,1997,16(2):13-14.
[34].师治贤,刘梅,杨月琴,等.糖茶蔗种子脂肪酸含量分析[J].西北植物学报,2005,25(8): 1669-1671.
[35].王辉,李景明,张雪,等.洋葱中风味前体物质的气相色谱研究[J].食品工业科技,2006,27(6):174-176.
[36].于新蕊,丛月珠.大蒜的化学成分及其药理作用研究进展.中草药,1994,25(3):158-161.
[37].王辉,李景明,马钊,等.洋葱中含硫化合物的生理功效[J].食品工业科技,2005,26(5):187-189.
[38].江苏新医学院.中药大辞典[M].上海:上海人民出版社,1977.
[39]. Kameoka H, Iida H, Hashimoto S, et a1. Phytochemistry,1984,23:155.
[40]. Law Son L D, et al. HPLC法测定大蒜匀浆中的大蒜辣素及其他硫代亚磺酸酯含量[J].1992,7(5):209.
[41]. Angusti KT, Mathew PT . Experientia,1974,30:468.
[42]. Block E, Ahma S, Jain MK, et al . J Am Chem Soc,1984,104:8295.
[43]. Bernhard RA. Phytochemistry,1970,9:2019.
[44].贾江滨,刘静,谭亚非.大蒜化学成分研究进展[J].广东药学,1999,9(1):1-5.
[45]. Lawson LD, Wang ZY, Hughes BG. J Nat Prod,1991,54:436.
[46]. Mackenzie IA, Ferns DA. Phytochemistry,1977,16:763.
[47].何洪巨,唐晓伟,宋曙辉,等.韭葱挥发性物质的气相色谱[J].质谱分析质谱学报,2004,25:63-64.
[48].刘松忠,陈清,何洪巨,等.葱、蒜挥发性物质形成及影响因素研究进展[J].中国蔬菜,2005(增刊):35-38.
[49]. Schwimmer S,Weston W. Enzymatic development of pyruvic acid in onion as a measure of pungency. Agr Food Chem,1961,9:301-304.
[50]. Randle W M. Quantifying onion flavor compounds responding to sulfur fertility-sulfur increases levels of alk(en)yl cysteine sulfoxides and biosynthetic intermediates.Amer Soc HortSci,1995,120(6):1075 -1 081.
[51]. Coolong T W , Randle W M. Sulfur and nitrogen availability interact to affect the flavor biosynthetic pathway in onion.Amer Soc Hort Sci,2003,128(5):776-783.
[52]. Arthur Stoll, et al. Helv. Chim Acta.,1984,31:189;1432.
[53].许真,严永哲,卢钢,等.葱属蔬菜植物风味前体物质的合成途径及调节机制[J].细胞生物学杂志,2007,29:508-512.
[54]. Lancaster JE et a1. In:Rabinowitch H et a1.(eds.)Alliums and Allied Crops,Vol.III.Boca Raton,FL:CRC Press,1990,33.
[55]. Lawson LD.In:Koch HP et a1.(eds.)Garlic :The Science and Therapeutic Application of AIlium sativum L.and Related Species,2nd ed.,W illiares and W ilkins,Baltimore,USA,1996,37.
[56]. Eric Block, David Putman, Shu-Hai ZhaoAllium. Chemistry:GC-MS Analysis of Thiosulfinates and Related Compounds from Onion, Leek, Scallion, Shallot, Chive, and Chinese Chive[J]. Agric. Food Chem. 1992, 40, 2431-2436.
[57].钟国清.油脂碘值的测定方法研究[J].粮油食品科技,2004,12(1):29-30.
[58].洪滨,刘会洲.国内EPA及DHA研究现状和发展趋势[ J ].化工冶金, 1996, 17 (1) : 82– 841.
[59].吴冀华,裘爱冰.气相色谱法分析共轭亚油酸异构体[ J ].中国油脂, 2002, 27 (1) : 65– 671.
[60].陈亚飞,程速远,孙宇,等.毛细管气相色谱法测定海狗油胶丸中ω-3多不饱和脂肪酸的含量[J].药物分析杂志,2004,24(5):499-501.
[61].曹玉铭,王英爽,郭建春,等.气相色谱法测定黑加仑油中γ—亚麻酸、α一亚麻酸、亚油酸含量的研究[J].中国新医药,2004 3(3):52-53.
[62].邹月利,赵李霞,李冬梅.亚麻籽中不饱和脂肪酸含量的测定[J].生命科学仪器,2005,3(3):42-43.
[63].中华人民共和国国家标准GB/T 17376一1998.动植物油脂,脂肪酸甲醋制备.
[64].中华人民共和国国家标准GB/T 17377一1998.动植物油脂,脂肪酸甲醋的气相色谱分析.
[65]. Yeonhwa Park, Karen J. Albright, Zu Y. Cai, and Michael W. Pariza Comparison of Methylation Procedures for Conjugated Linoleic Acid and Artifact Formation by Commercial(Trimethylsilyl)diazomethane J. Agric. Food Chem. 2001, 49, 1158-1164
[66].可成友,吴晓芳.不饱和脂肪酸的气相色谱法同时测定[J].中国卫生检验杂志,2005,15(5):528-531.
[67].冯家力,潘振球,陈波.奶粉中EPA和DHA的衍生气相色谱分析[J].中华预防医学杂志, 1997, 31 (4) : 239– 2401.
[68].杨勇杰,姜瑞芝,陈英红,等.高其品猴头多糖中脂肪酸成分分析及含量测定[J].中成药,2005,27 (10):1196-1198.
[69].政玉梅,陈炳卿,杨艳梅,等.气相色谱法测定酸奶和奶油中c9,t11-共轭亚油酸的含量.中国卫生检验杂志,2002,12(1):6-7.
[70].马往校,段敏,孙新涛,等.定硫法测定大蒜中大蒜素含量及影响因素[J].天然产物研究与开发,2002 ,14 (6):22-23.
[71].黄志伟,黎中良,张宏志.玉林大蒜中大蒜素含量测定的研究[J].实验科学与技术,2006,3:22-23.
[72].张维勤.大蒜辣素的测定方法[J].山东农业科学,1988,5:49-52.
[73].甄志,李宇,陈鸿伟.Vario EL III元素分析仪测硫方法分析[J].电力科学与工程,2002,4:43-45.
[74].梁晓斌,奇文瑞.EA1110元素分析仪应用中存在的问题及对策[J].河南化工,2006,23(5):33-34.
[75].董德恩,李拓宏.用Vario.EL元素分析仪测定煤中碳等元素[J].同煤科技,2004,4:36,38.
[76].成力华.PE2400Ⅱ元素分析仪在石油添加剂硫含量测定中的应用[J].甘肃化工,1996,2:43-46.
[77].夏朝彬,黄芳.德国Heraeus CHN—O—S元素分析仪在油样分析中的应用[J].石油与天然气化工,1994,23(4):255-257.
[78].刘力,徐淑英.1106元素分析仪测硫方法的改进[J].分析仪器,1991,2:64-66.
[79].成力华.PE 2400Ⅱ元素分析仪在石油添加剂硫含量测定中的应用[J].甘肃化工,1996,2:43-46.
[80].邓广勇.再谈CHNS元素分析仪测定硫含量的问题[J].分析仪器,2000,4:50-52.
[1].杭晓敏,唐涌濂,柳向龙.多不饱和脂肪酸的研究进展[J].生物工程进展,2001,21(4):18-21.
[2].王利华,霍贵成.ω-3不饱和脂肪酸的生物学作用[J].东北农业大学学报,2001,32(5):100-104.
[3]. Derberg, J. Nutr, Rev. 44:125-1 34,1986
[4]. Lands, W. E.M..In Health Effects of Polyunsaturated Fatty Acids in Seafood,. Academic P, 1986, pp:319-352
[5]. Yongmanitchai, Wichien, Ward, et al. Omega-3 fatty acids: alternative sources of production[J]. Pro. Biochem,1989,24(4):117-125.
[6]. Ratledge, C. in, Technological Advances in Improved and Altemative Sources of Lipid, edited by Katuda, Y., Blackre Academic,1994, pp, 235-291.
[7]. Ratledge Colin. Single cell oils - have they a biotechnological future[J]? Trends in Biotechnology,1993,11(7):278-84.
[8]. Molina Gnma, E., Danchez Perez, J. A. Appl. Microbiol. Biotechnol, 4l,23-27,1994.
[9].尤新.功能性发酵食品[M].北京:中国轻工业出版社,2000.
[10].马玉平,李德海,王忠萍.多不饱和脂肪酸的生物合成[J].中国乳品工业,2002,30(2):27-30.
[11].宋思扬,楼士林.生物技术概念[M].北京:科学出版社,1999.
[12].刘世杰,宋代军.多不饱和脂肪酸的营养研究进展[J].饲料博览,2002,2:11-13.
[13].顾关云,王欢,蒋昱.国外医药.植物药分册,2005,20(6):231-236.
[14].刘育葵.月见草与γ-亚麻酸[J].宁夏医学杂志,1995,17(5):314.
[15].赵占英.月见草不同品系种子油的成分测定[J].中国野生植物资源,2000,16(4):29.
[16].曹毅,赵冬梅.月见草油的制备、功能和应用[J].粮油食品科技,1999,7(3):17—20.
[17].仲玉梅.营养素:γ-亚麻酸[J].食品工业科技,1990,(2):32—36.
[18].朗维,刘柏.复方月见草油降低花样滑冰等项目女运动员体脂含量提高心脏泵血功能的研究[J].冰雪运动,1996,(4):35—39.
[19].汪开治.妇科良药月见草[J].植物杂志,2002,(6):14.
[20].朱健,赵玲爱,朱鸣.一种颇有开发价值的药用油料植物-月见草[J].陕西林业科技,1999,(1):27-28.
[21]. Fukushima, Michihiro,Ohhashi, et al. Effects of diets enriched in n-6 or n-3 fatty acids on cholesterol metabolism in older rats chronically fed a cholesterol-enriched diet[J].Lipids,2001,36(3):261-266.
[22]. De La Cruz J P; Martin-Romero M; Carmona J A; Villalobos M A; Sanchez de la Cuesta F Effect of evening primrose oil on platelet aggregation in rabbits fed an atherogenic diet. Thromb Res,1997,87(1):141.
[23]. Villalobos M. A., De La Cruz J. P., Martin-Romero M., et al. Effect of dietary supplementation with evening primrose oil on vascular thrombogenesis in hyperlipemic rabbits[J]. Thrombosis and Haemostasis,1998,80(4):696-701.
[24]. Snuth. W. L. and Borgeat. P., in Biochemistry of Lipid and Membranes, edited by Vance. D.E. and Vance .J. E., Benjamin Cuinmings, Redwood City. CA. USA,1985:325-360.
[25]. Grammatikos S I,Subbaiah P V,Victor T A,et al. Diverse effects of essential (n-6 and n-3) fatty acids on cultured cells[J]. Cytotechnology ,1994,15(1-3):31-50.
[26]. Marahall. A. and Van Elswyk. M. E., Nutr. Today 30,207-213,1995.
[27]. Arthur Sto11. Hely.Chilift.AcIa.,I948,31(189):1432
[28]. Schwimmer S,Weston W.Enzymatic development of pyruvic acid in onion as a measure of pungency.Agr Food Chem,1961,9:301-304.
[29]. Randle W M.Quantifying onion flavor compounds responding to sulfur fertility-sulfur increases levels of alk(en)yl eysteine sulfoxides and biosynthetic intermediates. Amer Soc Hort Sci,1995,120(6):1075 -1081.
[30]. Colong T W, Randle W M. Sulfur and nitrogen availability interact to affect the flavor biosynthetic pathway in onion. Amer Soc Hort Sci,2003,128(5):776-783.
[31]. Freeman, George G.; Whenham, et a1. Volatile components of Allium species in terms of S-alk(en)yl-L-cysteine sulfoxides present as flavor precursors. J.Sci Food Agric,1975,26(12):1869-86.
[33]. Bianchini F, Vainio H. Allium vegetables and organsulfur compounds: do they help prevent cancer?[J]. Environ Health Perspect,2001,109(9):893-902.
[34].上海第二制药厂.中草药通迅,1976,(10):8有效的活性化合物将会被发现
[35]. Karrer W, et al. Constitution and Vorkonmmen der organosschen Planzenstoffe (exclusive Alla oids).1976
[36].丁智慧.云南植物研究,1988,10(2):223.
[37]. Stoll, A.; Seebeck, E. The isolation of sinigrin as the true, crystallized mother substance of horse-radish oil[J]. Helvetica Chimica Acta,1948,31:1432-4.
[40]. Randle W M .Increasing nitrogen concentration in hydroponic solutions affects onion flavor and bulb quality.Amer Sac Hort Sci,2000,125(2):254-259.
[42].张银华.除臭大蒜口服液的GC/MS分析[J].天然产物研究与开发, 1997,(2):53-57.
[43]. Mackenzie,Ian A,Ferns, et al. The composition of volatiles from different parts of Allium tuberosum plants[J]. Phytochemistry,1977,16:763-4.
[44]. a Tahara, Satoshi; Okamura, Hiroyuki; Miura, Yuzo; Mizutani, Junya. Hydantoin derivatives of S-alk(en)yl-L-cysteine-S-oxides. Part IV. -Elimination reaction of antimicrobial L-5-alk(en) ylthiomethylhydantoin-S-oxides. Agric boil chem.,1979,43(10):2017. b Tahara, Satoshi; Mizutani, Junya. Hydantoin derivatives of S-alk(en)yl-L-cysteine-S-oxides. Part V. L-5-Alk(en)ylthiomethylhydantoin-( )-S-oxides: non-enzymical precursors of fresh flavors of Allium plants. Agric boil chem.,1979,43(10):202l.
[46].鲁信元.从大蒜中提取蒜油[J].中国调味品,1990,7:18-1.
[47]. Block E, Gulati H, Putman D, Sha D, You N, Shu—Hai, Z.Allium chemistry:Synthesis of 1-[alk(en)yl sulfinyl] propyl alk(en)yl disulfides (cepaenes),antithrombotic flavorants from homogenates of onion (Allium cepa)[J]. J Agric Food Chem,1997,45: 4414-4422.
[48].嚋忠梅,于德泉,丛浦珠.葱属植物化学及药理研究进展[J].药学学报,1999,34(5):395-400.
[49]. Lukes T M. Thin-layer chromatography of cysteine derivatives of onion flavor compounds and the lacrimatory[J].J Food Sci,1971, 36:662.
[50]. Virtanen A I,Matikkala E J. The isolation of S-methlcysteine sulfoxide and S—n-propyl cysteine sulfoxide from onion (Allium Cepa) and the antibiotic activity of crushed onion[J]. Acta ChemStand,1959,13:1898.
[51]. Kohhof T M, Stricks W, Kapoor R C.Equilibrium constants of exchange reactions of cystine with glutathione and with thioglyeolic acid both in the oxidized and reduced state[J]. J Am Chem Soc,1955,77:4733.
[52]. Kamini E N,Syeda A. Antimicrobial activity of garlic and onion extracts [J]. Pharmazie,1983,38(11):747-748.
[53]. Kim J. Anti-bacterial action of onion extracts against oral pathogenic bacteria [J]. J Nihon Univ Sch Dent,1997,39( 3):l36-141.
[54]. Naganawa R, Iwata N, Ishikawa K, et al. Inhibition of microbial growth by ajoene, a sulfur-containing derived from garlic [J]. Appl Environ Microbiol, 1996, 62(11):4238-4242.
[55]. Mathew P T, Augusti K T. Isolation of hypo—and hyperglycemic agents from Allium Cepa Linn[J]. Indian J Exp Biol,1973,1 1:573.
[56]. Augnsti K T. Chromatographic identification of certain sulfoxides of cysteine present in onion (Allium Cepa Linn) extract[J]. Curr Sci,1976,45:863.
[57]. Katzen H M, Tietz F. Studies on the specificitv and mechanism of action of hepatic glutathione insulin transhydrogenase [J]. J Biol Chem,1966,241:3561.
[58]. Black S. Reduction of sulfoxide and disulfides, in Methods in Enzymology[M].New York:Academic Press,l962,992.
[59]. Augusfi KT. Gas chromatographic analysis of onion principles and a study on their hypoglycemic action[J] . Indian J Exp Biol,1976,14:110.
[60]. Augnsti K T, Roy V C M, Semple M. Effect of allyl propyl disulphide isolated from onion (Allium Cepa L) on glucose tolerance of alloxan diabetic rabbits[J]. Experimentia,1974,30:l l9.
[61]. Augnsti K T, Benaim M E. Effect of essentia1 oil of onion(allyl propyl disulpbide) on blood glucose, free fatty acid and insulin levels of normal subjects[J]. Clin Chim Acta, 1975, 60:l21.
[62]. Wilcox B F, Joseph P K, Augusti K T. Effects of allyl propyl disulphide isolated from Allium Cepa Linn on high-fat fed rats . Indian J Exp Biol, 1973(11):239.
[63]. Sheela C G,Augusti K T. Antidiabetic effects of S—allyl cystenine Sulphoxide isolated fromgarlic and onion[J]. India Journal of Experimental Biology,1992,30(6):523-526.
[64]. Sheela C,Kumud K,Augusti K. Anti—diabetic effects of onion and garlic sulfoxide amino acids in rats [J]. Planta Med,1995,61:356-357.
[65]. Ariga T, Tsuj K, Seki T, et al. Antihrombotic and antineoplastic effects of phyto-organosulfur compounds [J]. Biofactors, 2000, 13(1-4):251-255.
[66].严常开,曾繁典.大蒜的主要化学成分及其药理作用的研究进展[J].中国新药杂志,2004,13(8):688-691.
[67]. Block B E. The organosulfur chemistry of the genus Allium implications for the organic chemistry of sulfur [J].Angew Chem Int Eds Engl,1992,31:1135-1178.
[68]. MacDonald JA, Langler RF. Structure-activity relationships for selected sulfur-rich antithrombotic compounds[J]. Biochem Biophys Res Commun, 2000,273(2):421-424.
[69]. Itokawa Y, Inoue K, Sasagawa S, Fujiwara M. Effect of S-methyl cysteine sulphoxide, S-allyl cysteine sulfoxide and related sulfur containing amino acids on lipid metabolism of experimental hypercholesterolemic rats[J]. J Nutr,1973,103:88.
[70]. Hatono S, Jimenez A, Wargovich M. Chemopreventive effect of S-allylcysteine and the relationship to the detoxification enzyme glutathione S-transferase[J]. Carcinogenesis, 1996,17(5): 104l- 1044.
[71]. Steerenberg P A, et a1. Protection of UV—induced suppression of skin contact hypeI.sensitivity: A common feature of flavonoids after oral administration [J]. Photochemistry and Photobiology ,1998, 67(4):456-461.
[72]. Franca B,Harri V. Allium vegetables and organosulfur compounds: do they help prevent cancer [J]. Environmental Health Perspectives,2001,109(9):893-901.
[73]. Hatono S, Jimenez A, Wargovich M. Chemopreventive effect of S-allylcysteine and the relationship to the detoxification enzyme glutathione S-transferase[J]. Carcinogenesis, 1996,17(5): 104l-1044.
[74]. Steerenberg P A. Protection of UV—induced suppression of skin contact hypersensitivity: A common feature of flavonoids after oral administration [J]. Photochemistry andPhotobiology ,1998,67(4):456-461.
[75]. Franca B, Harri V. Allium vegetables and organosulfur compounds: do they help prevent cancer [J]. Environmental Health Perspectives,2001,109(9):893-901.
[76].Bose C, Guo J, Zimniak L, et al. Critical role of allyl groups and disulfide chain in induction of Pi class glutathione transferase in mouse tissues in vivo by diallyl disulfide, naturally occurring hemopreventive agent in garlic[J].Carcinogenesis, 2002,23(10):1661-1665.
[77].中华人民共和国药典(二部)[Z].北京:化学工业出版社,2000.
[78].赵永凤,张胜义.示波电位滴定法测定含硫药物[J].理化检验:化学分册,1994,30(6):353-355.
[79].林战英,许东航,李新,等.含硫药物的氧瓶燃烧一电位滴定法测定[J].现代应用药学,1994,11(5):41.
[80].施先义.碘量法测定邻甲基苯硫脲[J].理化检验:化学分册.2003,39(3):168-169.
[81].张银华.除臭大蒜口服液的GC/MS分析[J].天然产物研究与开发,1997,9(2):53-57.
[82].云南植物研究,1988,10(2):223.
[84].吴丽明,邱光清.大蒜精降血脂和毒性实验研究[J].中成药,1997,19(12),30-31.
[85]. Martin-Lagos, R. Artacho; Serrano, M. F. Olea; Lopez, M. D. Ruiz.Determination of organic sulfur compounds in garlic extracts by gas chromatography and mass spectrometry[J].Food Chem 1995,53(1):9l-3.
[86]. Vernin Gaston,Metzger Jacques,Fraisse Daniel,et al.GC-MS (EI, PCI, NCI) computer analysis of volatile sulfur compounds in garlic essential oils[J]. Application of the mass fragmentometry SIM technique. Planta Med 1986,52:96-101.
[87]. Block, Eric; Iyer, Rajeshwari; Grisoni, Serge; et al .Lipoxygenase inhibitors from the essential oil of garlic. Markovnikov addition of the allyldithio radical to olefins[J]. J Am Chem Soc 1988,110:7813.
[88].Djordjevic Mirjana V.,Gay Scott L., Bush Lowell P, et al.Tobacco-specific nitrosamine accumulation and distribution in flue-cured tobacco alkaloid isolines[J]. Agric.Food Chem,1989.37:752-6.
[89]. Yu Tung Hsi,Wu Chung May,Liou Yoh Cherng. Effects of pH adjustment and subsequent heattreatment on the formation of volatile compounds of garlic[J].Journal of Food Sci,54(3):632-5.
[90]. Vernin, Gaston; Metzger, Jacques; Fraisse, Daniel; Scharff, Claude. GC-MS (EI, PCI, NCI) computer analysis of volatile sulfur compounds in garlic essential oils.Application of the mass frag mentometry SIM technique.Planta Med, 1986,52:96.
[91]. Beiner K, Popp P, Wermrich R, et a1. Determination of volatile sulfur compounds in water samples by GC-MS with selective preconcentration[J]. J Chromagr A,2002,968(1):171-176.
[92].第二次全国质谱学会议资料选编.北京:原子能出版社,1982:115.
[93].分析测试学报,1990,9(3):11.
[96]. Laakso I., Seppanen-Laakso T,Hiltunen R,et al. Volatile garlic odor components: gas phases and adsorbed exhaled air analyzed by headspace gas chromatography-mass spectrometry[J]. Planta Med,1989,55(3):257-61.
[97]. Nielsen A T, Jonsson S. Trace determination of volatile sulfur compounds by solid-phase microextraction and GC—MAS[J]. Analyst, 2002, 127: 1045—1049.
[98]. Giacomo Mauriello, Rosanna Marino, Mauriz]o D’Auria, et a1. De termination of volatile compounds from truffles via SPME-GC-MS [J]. J Chromatogr Sci, 2004, 42(6): 299-355.
[99].GryglewiczG, Gryglewicz S. Determination of elemental sulfur in coal by gas chromatography-mas spectrometry [J]. Anal Bioanal Chem,2001,370(1):60-63.
[100]. Moreira N, Guedes de Hnho P, Vasconcelos I. Method for analysis of heavy sulphur compounds using gas chromatography with flame photometric detection[J]. Anal Chim Acta,2004, 513(1):183-189.
[101]. Chambers L, Duffy M L. Determination of total and sepciated sulfur content in petrochemical samples using a pulsed flam photometric detector [J]. J Chromatogr Sci,2003,41(10):528-534.
[102]. Moehizuki Emiko. J-Chromatogr, 1988,455-271.
[103].食品科学1996,23(1):107.
[104].中草药,1994,25(10):517-518.
[105].赵淑杰,李玲玲.复方滴鼻剂制备工艺的优选及大蒜素稳定性的考察中国中药杂志,1994,19(4):227-229.
[106]. Lawson Larry D,Wood Steven G,Hughes Bronwyn G. HPLC analysis of allicin and other thiosulfinates in garlic clove homogenates[J]. Planta Medica,1991,57(3),263-70.
[107]. El-Tanbouly, Nebal D.; Abdel-Megid, Roufaydah M. HPLC determination of alliin in garlic and garlic products and their antimicrobial activity.J Pharm Sci,1994,3(3A):120.
[108]. Usui Teruyuki,Suzuki Seiji. Isolation and identification of antiallergic substances from garlic (Allium sativum L.)[J]. Natural Medicines,1996,50(2):135-137.
[109]. Davis Larry E,Shen Jinkun,Royer Robert E. In vitro synergism of concentrated Allium sativum extract and amphotericin B against Cryptococcus neoformans[J]. Planta Medica,1994,60(6):546-9.
[110]. F. J Kappenberg, et a1. Pharm. Zig. Wiss, 1990,l35:189.
[111]. Fujiwara Motonori,Yoshimura Masao,Tsuno Sadako. Allithiamine, a newly found derivative of vitamin B1. III. Allicin homologs in Allium plants[J]. Journal of Biochemistry (Tokyo, Japan),1955, 42: 591-601.
[113]. Lancaster Jane E,Kelly Kathleen E. Quantitative analysis of the S-alk(en)yl-L-cysteine sulfoxides in onion (Allium cepa L.)[J]. Journal of the Science of Food and Agriculture,1983,34(11):1229-35.
[114]. Nigalna Ekkad,Calvin 0 Huber. A coulometer for determination of residual sulfite following the dechlorination of water[J]. Anal Chim Acta,1996,332(23):155-160.
[115]. Ibrahim I, Cemal Y, Humeyra B. Construction and response characteristics of a sulfite/hydrogen sulfite-selective all-solid state contact electrode based on the 4-methylpiper -idinedithiocarbamite complex of mecury (Ⅱ)[J]. Analyst,1996,121:1873-1876.
[116]. Xu Y J, Lu C Y, Chen K, et a1. A conductance sensor for dissolved sulphur dioxide using a senes piezoelectric crystal device[J]. Talanta,1996,43(8):1297-1303.
[117]. Yang J H, Yang P H, Meng G Y. A fully solid state SOx(x= 2,3)gas sensor utilizing Ag—β一alumina as solid electrolyte[J]. Sens Aauators,1996,31B(3):209-212.
[118]. Chiou C Y, Chou T C. Ampemmetric sulfur dioxide sensors using the gold deposited gas-diffusion electrode [ J ]. Electroandysis,1996,8(12):1179-1182.
[119]. Mexiner F, Jaeschke W. Detection of low atmospheric sulfur dioxide concentration with achemiluminescence technique[J]. Int J Environ Anal Chem,1981,10(1):51-67.
[120]. Lin Jin—Ming,Hobo To~hiyuk1. Flow-injection analysis with chemiluminescent detection of sulphite using Na2CO3一NaHCO3-Cu2+ system[J]. Anal Chim Acta,1996,323(1/3):69-74.
[121]. Huang Yu—Liang, Kim Jong Min, Schmid R D. Determination of sulphite in wine through flow-injection analysis based on the suppression of luminol chemiluminescence [J].Anal Acta,1992,266(2):317-323.
[122].朱国辉,朱庆枝,许金钩.荧光光度法测定大气中痕量二氧化硫[J].分析化学,1999,27(11):1303-1305.
[123].陈巧珍,胡克季,三浦恭之.间接荧光检测离子色谱法分析维生素C、亚硫酸根和硫代硫酸根[J].色谱,1999,17(5):480-482.
[124].马往校,段敏,孙新涛,等.定硫法测定大蒜中大蒜辣素含量及影响因素[J].天然产物研究与开发,2002,14(6):22-23.
[125].张维勤.大蒜辣素的测定方法[J].山东农业科学,1988,5:49-52.
[126].朱英贵,王伦,赵丹华,等.氢氧化钠化学发光法测定痕量硫[J].分析实验室,2003,22(4):72-74.
[127]. E.Block. The organosulfur chemistry of the genus Allium -Implications for the organic chemistryofsulfru[J]. Angew.Chem,1992,31(9):l135-l178.
[128].周重洋,陈海波,罗燕,等.用生物检定法测定大蒜油中大蒜辣素和大蒜新素的含量[J].中草药,1997,28(1):18-20.