蜂王浆中四环素族药物残留测定方法研究
|
摘要
蜂王浆是由哺育蜂头部咽下腺和上颚腺共同分泌的粘性物质,是蜂王幼虫正常发育的必需食物。蜂王浆作为保健和功能食品,具有多方面的功效,长期食用有益于人体健康。中国是蜂王浆的主要生产和出口国家,主要出口到欧洲,日本和美国。随着蜂王浆产品消费的增加,蜂王浆中的药物残留逐渐引起了人们的关注。此外,随着中国蜂王浆出口量的增加,欧盟,美国和日本等国为了保护本国的贸易利益,设置了许多技术壁垒。这些国家和组织制订了苛刻的药物残留限量,建立相应的检测方法来限制中国蜂王浆产品出口。常规的测定方法很难满足这些国家和组织的限量要求。目前,我国并没有相应的检测蜂王浆中四环素的标准方法。为了保证我国蜂王浆产品的正常出口,建立简单快速测定蜂王浆中的四环素类药物残留的方法是非常必要和有意义的。
尽管已经报道了许多测定四环素类药物的方法,但这些方法并不适合测定蜂王浆中的四环素族残留,因为蜂王浆基质比其他食品更复杂,需要建立新的测定方法来监控蜂王浆中四环素族残留。在本研究中,我们制订了LC/MS法测定蜂王浆中的四环素残留。蜂王浆样品中的四环素药物残留用0.1mol/L Na2EDTA-Mcllvaine缓冲溶液提取,经过Oasis HLB柱净化,然后采用LC/MS进行测定。本方法的加标回收率在65.0%～86.8%之间,相对标准偏差在5.53%～11.30%之间,对蜂王浆中土霉素、四环素的检出限小于10 ng/g,金霉素、多烯环素的检出限小于20 ng/g。方法的各项指标符合兽药残留的要求。方法经过超高效液相色谱串联质谱的验证,证明方法有很好的适用性。方法经过不同实验室的复核,各项参数符合实验室间方法比对的各项要求,方法经过审定,形成相应的国家标准。
应用本研究制订的标准方法对我国蜂王浆的主要生产区的一些王浆样品进行检测,抽检的样品中有四环素类药物检出,主要检出的药物为土霉素,金霉素,说明我国蜂王浆产品中四环素族残留存在潜在威胁。
Royal jelly (RJ) is a viscous substance secreted by the hypopharyngeal and mandibular glands of nurse worker bees (Apis mellifera L.) that constitutes the essential food for the larvae of the queen bee.In modern diets, the use of RJ has significantly increased because of the trend toward use of healthy, organic, nutraceutical, and functional foods, and dietary supplements. The peoples’s republic of China is the world’s largest producer and exporter of royal jelly. Most of the Chinese RJ production is distributed in Europe, Japan, and the United States. As a result of the increasing interest in RJ in human health, the concern of drugs residues in royal jelly is also increasing. In addition, with export quantity of RJ continuously increasing, EU, USA and Japan made lots of TBs (technological barriers) in order to protect native trade benefits. Some tough MRL (maximum residue limit) and relative testing methods was established by EU, USA and Japan. It is difficult for common methods to analysis tetracyclines in RJ. Nowdays, there is no standard methods available for determination of tetracyclines in RJ in our country. Development of simple and rapid methods for determination of tetracyclines in RJ is therefore necessary and would be valuable for export trade of RJ.
Although a number of methods have been reported for determination of tetracyclines in different tissues and products, these methods can not be suitable to monitoring tetracyclines in RJ because of the matrix of RJ is more complex than others’s. In this study, we developed a LC/MS method for determination of tetracyclines in RJ. Tetetracycline antibiotic residues were extracted from royal jelly samples using a buffer containing 0.1mol/L Na2EDTA-Mcllvaine.The sample extracts were further purified and enriched by Oasis HLB.The recoveries of four tetracyclines ranged from 65.0 to 86.8% in fortified blanks, the relative standard deviations were between 5.53 % and 11.30 %. The method detection limits of oxytetracycline and teracycline were <10μg/kg, chlortetracycline and doxycycline were <20μg/kg, respectively. These results indicated the method meet the demand of animal drugs residue analysis. The developed method was validated by ultra-performance liquid-MS-MS and other laboratories. After examination the method developed national determination method standard.
Some samples collected from the main RJ produce areas were detected using the developed method. Some positive samples were found among which oxytetracycline and chlortracycline were main detectable drugs, which means it has a potential threat in RJ consumption.
尽管已经报道了许多测定四环素类药物的方法,但这些方法并不适合测定蜂王浆中的四环素族残留,因为蜂王浆基质比其他食品更复杂,需要建立新的测定方法来监控蜂王浆中四环素族残留。在本研究中,我们制订了LC/MS法测定蜂王浆中的四环素残留。蜂王浆样品中的四环素药物残留用0.1mol/L Na2EDTA-Mcllvaine缓冲溶液提取,经过Oasis HLB柱净化,然后采用LC/MS进行测定。本方法的加标回收率在65.0%～86.8%之间,相对标准偏差在5.53%～11.30%之间,对蜂王浆中土霉素、四环素的检出限小于10 ng/g,金霉素、多烯环素的检出限小于20 ng/g。方法的各项指标符合兽药残留的要求。方法经过超高效液相色谱串联质谱的验证,证明方法有很好的适用性。方法经过不同实验室的复核,各项参数符合实验室间方法比对的各项要求,方法经过审定,形成相应的国家标准。
应用本研究制订的标准方法对我国蜂王浆的主要生产区的一些王浆样品进行检测,抽检的样品中有四环素类药物检出,主要检出的药物为土霉素,金霉素,说明我国蜂王浆产品中四环素族残留存在潜在威胁。
Royal jelly (RJ) is a viscous substance secreted by the hypopharyngeal and mandibular glands of nurse worker bees (Apis mellifera L.) that constitutes the essential food for the larvae of the queen bee.In modern diets, the use of RJ has significantly increased because of the trend toward use of healthy, organic, nutraceutical, and functional foods, and dietary supplements. The peoples’s republic of China is the world’s largest producer and exporter of royal jelly. Most of the Chinese RJ production is distributed in Europe, Japan, and the United States. As a result of the increasing interest in RJ in human health, the concern of drugs residues in royal jelly is also increasing. In addition, with export quantity of RJ continuously increasing, EU, USA and Japan made lots of TBs (technological barriers) in order to protect native trade benefits. Some tough MRL (maximum residue limit) and relative testing methods was established by EU, USA and Japan. It is difficult for common methods to analysis tetracyclines in RJ. Nowdays, there is no standard methods available for determination of tetracyclines in RJ in our country. Development of simple and rapid methods for determination of tetracyclines in RJ is therefore necessary and would be valuable for export trade of RJ.
Although a number of methods have been reported for determination of tetracyclines in different tissues and products, these methods can not be suitable to monitoring tetracyclines in RJ because of the matrix of RJ is more complex than others’s. In this study, we developed a LC/MS method for determination of tetracyclines in RJ. Tetetracycline antibiotic residues were extracted from royal jelly samples using a buffer containing 0.1mol/L Na2EDTA-Mcllvaine.The sample extracts were further purified and enriched by Oasis HLB.The recoveries of four tetracyclines ranged from 65.0 to 86.8% in fortified blanks, the relative standard deviations were between 5.53 % and 11.30 %. The method detection limits of oxytetracycline and teracycline were <10μg/kg, chlortetracycline and doxycycline were <20μg/kg, respectively. These results indicated the method meet the demand of animal drugs residue analysis. The developed method was validated by ultra-performance liquid-MS-MS and other laboratories. After examination the method developed national determination method standard.
Some samples collected from the main RJ produce areas were detected using the developed method. Some positive samples were found among which oxytetracycline and chlortracycline were main detectable drugs, which means it has a potential threat in RJ consumption.
引文
1. 安 清聪,张 曦,李琦 华 .动物组 织 中 四环 素 类 抗生 素 残 留的 HPLC 检 测研究[J]. 动物医学进展,2005,26(6):92～94.
2. 薄海波,陈立仁.固相萃取法提取、高效液相色谱- 紫外检测法同时测定蜂王浆中土霉素、四环素、金霉素残留[ J ]. 中国卫生检验杂志, 2004,14 (6) : 699～ 700.
3. 蔡志斌,张英, 刘丽.固相萃取- 高效液相色谱法同时测定牛奶中 3 种四环素类抗生素残留量.中国卫生检验杂志,2007,17(2): 270~273.
4. 陈天豹,邓文汉,卢菀华.毛细管电泳法检测蜂蜜中残留的抗生素[J].色谱,2001, 19(1):91~93.
5. 戴 栗 洁 .土 霉 素 生 物 检 定 条 件 的 改 进 [J].锦 州 医 学 院 学 报 , 2001, 22(5): 34~37.
6. 冯淇辉,戎耀方,朱模忠.兽医临床药理学,科学出版社,1985.
7. 韩 胜明,何 薇莉,胡 长安,钱建华,李桂芬,冯 骞,陈广 全. 四 环素在 蜂群中的使用与蜂王浆中的残留. 中国养蜂,2003,54(4):34～35.
8. 黄志勇,蔡洪基,黄高凌.水产品中四环素类抗生素残留量的高效液相色谱测定方法[J].福建分析测试,2005,14(1):2093～2095.
9. 贾薇,孙璐,史向国.液相色谱- 质谱联用法测定牛奶中 4 种四环素类药物残留[J].沈阳药科大学学报,2002,19(2):96~100.
10. 蒋宏健,袁汉成.痕量四环素药物残余物液质联用分析及鉴定[J].现代仪器,2002, (1):35~36.
11. 李俊所,邱月明,王超.兽药残留分析,上海科学技术出版社,2002,369.
12. 李伟兵,蔡明明,宋依霖. 冻干 RJ 胶囊对肿瘤患者的免疫调节作用:附 30 例临床报告[J ] . 江苏中医,2000,21(2) :9~10.
13. 李亚琳,张俊升,王钦晖.酶联免疫法测定牛奶中四环素类药物的残留量[J].云南畜牧兽医,2005,2:27~30.
14. 李颖,李峰.HPLC 法同时测定禽兽组织中多种抗生素及克伦特罗残留的研究[ J ]中国卫生检验杂志, 2005, 15 (11) : 1321~13221.
15. 林 修光 , 寇 运同 ,贾 臻 .用 标 准 纸 片 法 快 速 测 定 鸡 肉 中 四 环 素 族 残 留 量 [J].口岸卫生控制,2001,6(3):19～20.
16. 林志彬. 70 年代以来 RJ 研究的进展[J ] . 蜜蜂杂志,1982,2:33~34.
17. 陆莉,林志彬. 蜂王浆的药理作用及相关活性成分的研究进展. 医药导报,2004, 23: 887~889.
18. 罗晓燕,林玉娜,刘莉治.SPE - HPLC 法同时测定水产品中四种抗生素残留的研究[ J ].现代预防医学,2005,32 (3) : 198~199.
19. 吕海涛,陈峰,姜悦.高效液相色谱法测定四环素类抗生药物中土霉素、强力霉素、四环素和金霉素.理化检验—化学分册,2007,43( 1):18~20.
20. 庞国芳,曹彦忠,张进杰.高效液相色谱法同时测定禽肉中土霉素、四环素、金霉素、强力霉素残留的研究[J].分析测试学报,2005,24(4):61～63.
21. 钱 英 .薄 层 色 谱 法 测 家 禽 肌 肉 组 织 中 的 四 环 素 和 土 霉 素 [J].职 业 与 健 康 ,2000, 16(12):37~40.
22. 秦皇岛出入境检验检疫局. GB/T 18932.23-2003-蜂蜜中土霉素、四环素 、金霉素、强力霉素残留量的测定方法液相色谱-串联质谱法. 北京:中华人民共和国质量监督检验总局,2003.
23. 秦皇岛出入境检验检疫局. GB/T 18932.4,2002-蜂蜜中土霉素、四环素 、金霉素、多烯环素残留量的测定方法-液相色谱法. 北京:中华人民共和国质量监督检验总局,2002.
24. 沈 洪刚 ,程 玲 玲 ,方 智 .酶 联 免 疫 法 快 速 测 定 猪 肉 中 四 环 素 族 残 留 量 [J].检验检疫科学,2000,10(2):42~43.
25. 石根勇,吕中明,陈新霞. RJ 冻干粉免疫调节作用的研究[J ] . 中国卫生检验杂志,2001 ,11(3) :306~307.
26. 宋 鹏 , 孙 彦 琴 , 潘 大 军 .牛 奶 中 四 环 素 类 抗 生 素 残 留 的 检 测 .河 南 畜 牧 兽 医综合版,2007,28 (5):41.
27. 谭 峰 .四 环 素 和 喹 诺 酮 类 药 物 分 析 方 法 研 究 [D].西 北 大 学 硕 士 学 位 论 文 ,2001, 5: 25~30.
28. 张国平,刘丛强,刘虹.固相萃取-色谱测定水、沉积物及土壤中氯霉素和 3种四环素类抗生素. 分析化学,31(2):315~319.
29. 张敬,戴秋萍,刘艺敏,等. RJ 冻干粉对小鼠免疫功能的影响[J ] . 上海铁道大学学报(医学版) ,2000 ,21 (9) :16~17.
30. 张敬,戴秋萍,刘艺敏等. RJ 冻干粉对小鼠肿瘤的抑制作用[J ] . 同济大学学报医学版,2001,22(5) :13~14.
31. 张兰,林子俺,谢增鸿.毛细管电泳用于水产品中五种抗生素的同时测定[J].分析测试技术与仪器,2004,10(1): 18~23.
32. 张素霞,李俊锁,钱传范.牛奶中四环素类药物多残留分析方法研究-MSPD- HPLC- UV[J].畜牧兽医学报, 2002, 33(1):51～ 54.
33. 张 子群,李 维,金宁. 应 用酶联 免疫技术检测动物 源性食品 中四环素类 抗生素残留的研究[J]. 黑龙江畜牧兽医, 2003,3:31～33.
34. 中华人民共和国国家进出口商品检验局. SN 0205,1993-出口蜂蜜中四环素族抗生素残留量检验方法杯碟法. 北京:中华人民共和国商业部,1993.
35. 中华人民共和国卫生部中国国家标准化管理委员会. GB/T5009.95-2003-蜂蜜中四环素族抗生素残留量的测定-微生物管碟法. 北京:中国国家标准化管理委员会,2003.
36. BlanchflowerWJ, McCracken RJ, Haggan AS. Confirmatory assay for the determination of tetracycline, oxytetracycline, chlortetracycline and its isomers in muscle and kindney using liquid chromatography - mass spectrometry [ J ].J Chromatogr B, 1997, 692 ( 1 ) : 351 ~361.
37. Canadian Food Inspection Agency .TetracycLine Residues in honey (hpLc-msd method ) ACC-042-v1.1, Canada, 2004.
38. Carson M C,Breslyn W. Simultaneous determination of multiple tetracycline residues in milk by metal chelate affinify chromatography [J].Journal of AOAC Internation at, 1996,79(l):2.
39. Casado-Terrones S, Segura-Carretero A, Busi S, Dinelli G, Fernández-Gutiérrez A. Determination of tetracycline residues in honey by CZE with ultraviolet absorbance detection.Electrophoresis,2007 , 28(16):2882~2887.
40. Catharina Y W A , wenhong L. A. bridging study between Liquid chromatography and microbial inhibition assay method for determining amoxicillin residues in catfish muscle [J].Journal of AOAC International, 1998, 81(1):3.
41. Choi KJ, Kim SG, Kim CW, Kim SH. Determination of antibiotic compounds in water by on-line SPE-LC/MSD.Chemosphere. 2007,66(6):977~984.
42. De Ruyck H, De Ridder H. Determination of tetracycline antibiotics in cow's milk by liquid chromatography/tandem mass spectrometry.Rapid Commun Mass Spectrom. 2007, 21(9):1511~1520.
43. De Wasch K, Okerman L, Croubels S.Detection of residues of tetracycline antibiotics in pork and chicken meat: correlation between results of screening and confirmatory tests [J].The Analyst, 1998, 123(12): 2737~2741.
44. Fujii A. Pharmacological effect of royal jelly [J]. Honeybee Science, 1995 ,16(1) :97 ~104.
45. Fujiwara S, Imai J, Yaeshima T, et al. A potent antibacterial protein in royal jelly. Purification and determination of the primary structure of royalisin[ J ] . J Biol Chem, 1990, 265(19):11333~11337.
46. Furusawa R. Liquid chromatographic determination of oxytetracycline in milk.J ChromatogrA, 1999, 839 (2): 247~251.
47. Gilliam, M, Argauer R. J. Oxytetracycline residues in surplus honey, brood nest honey and larvae after medication of colonies of honey bees, Apis mellifera, with antibiotic extender patties, sugar dusts, and syrup sprays. Enviro. Entomol 1981,10: 479~482.
48. Giulio S. Determination of sugars in royal jelly by HPLC. Apidologie 2006, 37: 84~90.
49. Gratacós-Cubarsí M, Fernandez-García A, Picouet P, Valero-Pamplona A, García-Regueiro JA, Castellari M. Formation of tetracycline degradation products in chicken and pig meat under different thermal processing conditions. J Agric Food Chem, 2007, 55(11):4610~4616.
50. Heggie, William, Galindro , eta1.Process for preparation of 4 - (des - dimethylamino) - tetracyclines. US.Patent 2000, 6143161.
51. Hisao O, Yoshitomo I, Norihisa K, Keiichi U, Masuo Y, Ken-Ichi H, Makoto S. Improvement of chemical analysis of antibiotics XII. Simultaneous analysis of seven tetracyclines in honey. J ChromatogrA 1987, 400: 253～ 261.
52. Hisao O, Yuko I, Hiroshi M. Chromatographic analysis of tetracycline antibiotics in foods. J ChromatogrA 2000, 393: 285-296.
53. Kamakura M, Mitant N, Fukuda T, etal. Antifatigue effect offresh roual jelly in mice. J Nutr Sci Vitaminol (Tokyo), 2001,47(6) :394 ~401.
54. Lee H J,Le M H, Ryu P D,et a1.Enzyme- linked immunosorbent assay for screening the plasma residues of tetracycline antibiotics in pigs [R].The Journal of veterinary medical science/the Japanese Society of Veterinary Science, 2001.63(5): 553~556.
55. LongAR, Hsieh LC, Malbrough MS, et al. Matrix solid - phase dispersion (MSPD) isolation and liquid chromatographic determination of oxytetracycline, tetracycline, and chlortetracycline in milk [ J ] 1J Assoc Off Anal Chem, 1990, 73 (3) : 379 ~ 384.
56. Marc C, Mario S, Siska C, et a1.Quantitative multi -residue analysis of tetracyclines and their 4- epimers in pig tissues by high - performance liquid chromatography combined with positive- ion electro spray ionization mass spectrometry [J].Analytica Chimica Acta.2003, 492 (1 - 2): 199~213.
57. Mascher A, LavagnoLi S, CuratoLo M . Determination of residual oxytetracycline in honey with an immunoassay kit. ApidoLogie,1996, 27: 229~233.
58. Matsui T, Yukiyoshi A, Doi S , et al . Gastrointestinal enzyme production of bioactive peptides from royal jelly protein and their antihypertensive ability in SHR[J ] . J Nultr Bioch 2002,13 (2) : 80 ~ 86.
59. Nagai T, Inoue, R. Preparation and the functional properties of water extract and alkaline extract of royal jelly. Food Chemistry 2004, 84, 181~186.
60. Nagai T, Sakai M, Inoue R. et al. Antioxidative activities of some commercially honeys, royal jelly and propolis [J]. Food Chemistry, 2001,75(2) :237~240.
61. Naoto Furusawa. An organic solvent - free method for determining oxytetracycline in COW ′ S milk [J].Solutions for separation scientists- LC/GC asia pacific, 2003, 6(3): 23～ 28.
62. Okamoto I, Taniguchi Y, Kunikata T, et al. Major royal jelly protein 3 modulates immune responses in vitro and in vivo[J]. Life Science 2003 ,73(16) :2029~2045.
63. Reybroeck W, Ooghe S, De Brabander H, Daeseleire E. Validation of the tetrasensor honey test kit for the screening of tetracyclines in honey J Agric Food Chem, 2007, 55(21):8359~8366.
64. Samanidou VF, Nikolaidou KI, Papadoyannis IN. Development and validation of an HPLC confirmatory method for the determination of seven tetracycline antibiotics residues in milk according to the European Union Decision 2002/657/EC.J Sep Sci. 2007 (15):2430~2439.
65. Santiago Valverde R, Sánchez Pérez I, Franceschelli F, Martínez Galera M, Gil García MD. Determination of photoirradiated tetracyclines in water by high-performance liquid chromatography with chemiluminescence detection based reaction of rhodamine B with cerium (IV).J Chromatogr A, 2007,1167(1):85~94.
66. Scheer M. Studies on the antimicrobial agents against five pathogens isolated in calves in the Netherlands [J].VetQuart.1900.12( 4) : 212～ 219.
67. Schneider MJ, Braden SE, Reyes-Herrera I, Donoghue DJ.Simultaneous determination of fluoroquinolones and tetracyclines in chicken muscle using HPLC with fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2007,846(1-2):8~13.
68. Schneider MJ, Darwish AM, Freeman DW. Simultaneous multiresidue determination of tetracyclines and fluoroquinolones in catfish muscle using high performance liquid chromatography with fluorescence detection.Anal Chim Acta, 2007,586(1-2):269~274.
69. Smyrniotakis CG, Archontaki HA. C18columns for the simultaneousdetermination of oxytetracycline and its related substances by reversed-phase high performance liquid chromatography and UV detection.J Pharm Biomed Anal. 2007, 43(2):506~514.
70. Spisso BF, de Oliveira e Jesus AL, de Araújo MA Jr, Monteiro MA.Validation of a high-performance liquid chromatographic method with fluorescence detection for the simultaneous determination of tetracyclines residues in bovine milk.Anal Chim Acta, 2007, 581(1):108~117.
71. Stead SL, Caldow M, Sharma A, Ashwin HM, Sharman M, de-Rijk A, Stark J. New method for the rapid identification of tetracycline residues in foods of animal origin - using the PremiTest in combination with a metal ion chelation assay.Food Addit Contam,2007, J24(6):583-589.
72. Sun XY, Chen H, Gao H, Guo XQ. Screening of tetracycline residues in fish muscles by CCD camera-based solid-surface fluorescence. J Agric Food Chem, 2006, 54(26):9687~9695.
73. Sver L, Orsolic N, Tadic Z, et al. A royal jelly as a new potential immunomodulator in rats and mice[J] . Comp Immun Microbiol Infect Dis , 1996 ,19(1) :31~38.
74. Tamura T. Royal jelly from the standpoint of clinical pharmacology[J] . Honeybee Science 1985, 6(1) :117~124.
75. Traviesa-Alvarez JM, Costa-Fernández JM, Pereiro R, Sanz-Medel A. Direct screening of tetracyclines in water and bovine milk using room temperature phosphorescence detection.Anal Chim Acta, 2007,589(1):51~58.
76. Ueno R, Sangrungruang K, Miyakawa M.A simplifiedmethod for the determination of several fish drugs in edible fish and shrimp by high - performance liquid chromatography [J].Food Research Intemationa1.1999,32 (9):629～ 633.
77. Vega D, Agüí L, González-Cortés A, Yá?ez-Sede?o P, Pingarrón JM. Voltammetry and amperometric detection of tetracyclines at multi-wall carbon nanotube modified electrodes.Anal Bioanal Chem,2007 ,389(3):951~858.
78. Wang G Y, Lin ZB. Effects of 102hydroxy222decenoie acid on phagocytosis and cyktokines production of peritoneal macrophages in vitro [J ] . Acta Pharmacol Sin, 1997, 18 (2):180 ~182.
79. White CR. Optimization of a liquid chromatographic method for determination of oxytetracycline, tetracycline, and chlortetracycline in milk.J Assoc of Anal Chem Int, 1993, 76 (3): 549 ~554.
80. Ye Z, Weinberg HS, Meyer MT. Trace analysis of trimethoprim andsulfonamide, macrolide, quinolone, and tetracycline antibiotics in chlorinated drinking water using liquid chromatography electrospray tandem mass spectrometry.Anal Chem. 2007, 79(3):1135~1144.
81. Yoshitomo I, Hisao O, Norihisa K, Masuo Y, Ken-Ichi H, Makoto S. Improvement of chemical analysis of antibiotics : XIII. Systematic simultaneous analysis of residual tetracyclines in animal tissues using thin-layer and high-performance liquid chromatography J ChromatogrA 1987, 411: 313～ 323.
2. 薄海波,陈立仁.固相萃取法提取、高效液相色谱- 紫外检测法同时测定蜂王浆中土霉素、四环素、金霉素残留[ J ]. 中国卫生检验杂志, 2004,14 (6) : 699～ 700.
3. 蔡志斌,张英, 刘丽.固相萃取- 高效液相色谱法同时测定牛奶中 3 种四环素类抗生素残留量.中国卫生检验杂志,2007,17(2): 270~273.
4. 陈天豹,邓文汉,卢菀华.毛细管电泳法检测蜂蜜中残留的抗生素[J].色谱,2001, 19(1):91~93.
5. 戴 栗 洁 .土 霉 素 生 物 检 定 条 件 的 改 进 [J].锦 州 医 学 院 学 报 , 2001, 22(5): 34~37.
6. 冯淇辉,戎耀方,朱模忠.兽医临床药理学,科学出版社,1985.
7. 韩 胜明,何 薇莉,胡 长安,钱建华,李桂芬,冯 骞,陈广 全. 四 环素在 蜂群中的使用与蜂王浆中的残留. 中国养蜂,2003,54(4):34～35.
8. 黄志勇,蔡洪基,黄高凌.水产品中四环素类抗生素残留量的高效液相色谱测定方法[J].福建分析测试,2005,14(1):2093～2095.
9. 贾薇,孙璐,史向国.液相色谱- 质谱联用法测定牛奶中 4 种四环素类药物残留[J].沈阳药科大学学报,2002,19(2):96~100.
10. 蒋宏健,袁汉成.痕量四环素药物残余物液质联用分析及鉴定[J].现代仪器,2002, (1):35~36.
11. 李俊所,邱月明,王超.兽药残留分析,上海科学技术出版社,2002,369.
12. 李伟兵,蔡明明,宋依霖. 冻干 RJ 胶囊对肿瘤患者的免疫调节作用:附 30 例临床报告[J ] . 江苏中医,2000,21(2) :9~10.
13. 李亚琳,张俊升,王钦晖.酶联免疫法测定牛奶中四环素类药物的残留量[J].云南畜牧兽医,2005,2:27~30.
14. 李颖,李峰.HPLC 法同时测定禽兽组织中多种抗生素及克伦特罗残留的研究[ J ]中国卫生检验杂志, 2005, 15 (11) : 1321~13221.
15. 林 修光 , 寇 运同 ,贾 臻 .用 标 准 纸 片 法 快 速 测 定 鸡 肉 中 四 环 素 族 残 留 量 [J].口岸卫生控制,2001,6(3):19～20.
16. 林志彬. 70 年代以来 RJ 研究的进展[J ] . 蜜蜂杂志,1982,2:33~34.
17. 陆莉,林志彬. 蜂王浆的药理作用及相关活性成分的研究进展. 医药导报,2004, 23: 887~889.
18. 罗晓燕,林玉娜,刘莉治.SPE - HPLC 法同时测定水产品中四种抗生素残留的研究[ J ].现代预防医学,2005,32 (3) : 198~199.
19. 吕海涛,陈峰,姜悦.高效液相色谱法测定四环素类抗生药物中土霉素、强力霉素、四环素和金霉素.理化检验—化学分册,2007,43( 1):18~20.
20. 庞国芳,曹彦忠,张进杰.高效液相色谱法同时测定禽肉中土霉素、四环素、金霉素、强力霉素残留的研究[J].分析测试学报,2005,24(4):61～63.
21. 钱 英 .薄 层 色 谱 法 测 家 禽 肌 肉 组 织 中 的 四 环 素 和 土 霉 素 [J].职 业 与 健 康 ,2000, 16(12):37~40.
22. 秦皇岛出入境检验检疫局. GB/T 18932.23-2003-蜂蜜中土霉素、四环素 、金霉素、强力霉素残留量的测定方法液相色谱-串联质谱法. 北京:中华人民共和国质量监督检验总局,2003.
23. 秦皇岛出入境检验检疫局. GB/T 18932.4,2002-蜂蜜中土霉素、四环素 、金霉素、多烯环素残留量的测定方法-液相色谱法. 北京:中华人民共和国质量监督检验总局,2002.
24. 沈 洪刚 ,程 玲 玲 ,方 智 .酶 联 免 疫 法 快 速 测 定 猪 肉 中 四 环 素 族 残 留 量 [J].检验检疫科学,2000,10(2):42~43.
25. 石根勇,吕中明,陈新霞. RJ 冻干粉免疫调节作用的研究[J ] . 中国卫生检验杂志,2001 ,11(3) :306~307.
26. 宋 鹏 , 孙 彦 琴 , 潘 大 军 .牛 奶 中 四 环 素 类 抗 生 素 残 留 的 检 测 .河 南 畜 牧 兽 医综合版,2007,28 (5):41.
27. 谭 峰 .四 环 素 和 喹 诺 酮 类 药 物 分 析 方 法 研 究 [D].西 北 大 学 硕 士 学 位 论 文 ,2001, 5: 25~30.
28. 张国平,刘丛强,刘虹.固相萃取-色谱测定水、沉积物及土壤中氯霉素和 3种四环素类抗生素. 分析化学,31(2):315~319.
29. 张敬,戴秋萍,刘艺敏,等. RJ 冻干粉对小鼠免疫功能的影响[J ] . 上海铁道大学学报(医学版) ,2000 ,21 (9) :16~17.
30. 张敬,戴秋萍,刘艺敏等. RJ 冻干粉对小鼠肿瘤的抑制作用[J ] . 同济大学学报医学版,2001,22(5) :13~14.
31. 张兰,林子俺,谢增鸿.毛细管电泳用于水产品中五种抗生素的同时测定[J].分析测试技术与仪器,2004,10(1): 18~23.
32. 张素霞,李俊锁,钱传范.牛奶中四环素类药物多残留分析方法研究-MSPD- HPLC- UV[J].畜牧兽医学报, 2002, 33(1):51～ 54.
33. 张 子群,李 维,金宁. 应 用酶联 免疫技术检测动物 源性食品 中四环素类 抗生素残留的研究[J]. 黑龙江畜牧兽医, 2003,3:31～33.
34. 中华人民共和国国家进出口商品检验局. SN 0205,1993-出口蜂蜜中四环素族抗生素残留量检验方法杯碟法. 北京:中华人民共和国商业部,1993.
35. 中华人民共和国卫生部中国国家标准化管理委员会. GB/T5009.95-2003-蜂蜜中四环素族抗生素残留量的测定-微生物管碟法. 北京:中国国家标准化管理委员会,2003.
36. BlanchflowerWJ, McCracken RJ, Haggan AS. Confirmatory assay for the determination of tetracycline, oxytetracycline, chlortetracycline and its isomers in muscle and kindney using liquid chromatography - mass spectrometry [ J ].J Chromatogr B, 1997, 692 ( 1 ) : 351 ~361.
37. Canadian Food Inspection Agency .TetracycLine Residues in honey (hpLc-msd method ) ACC-042-v1.1, Canada, 2004.
38. Carson M C,Breslyn W. Simultaneous determination of multiple tetracycline residues in milk by metal chelate affinify chromatography [J].Journal of AOAC Internation at, 1996,79(l):2.
39. Casado-Terrones S, Segura-Carretero A, Busi S, Dinelli G, Fernández-Gutiérrez A. Determination of tetracycline residues in honey by CZE with ultraviolet absorbance detection.Electrophoresis,2007 , 28(16):2882~2887.
40. Catharina Y W A , wenhong L. A. bridging study between Liquid chromatography and microbial inhibition assay method for determining amoxicillin residues in catfish muscle [J].Journal of AOAC International, 1998, 81(1):3.
41. Choi KJ, Kim SG, Kim CW, Kim SH. Determination of antibiotic compounds in water by on-line SPE-LC/MSD.Chemosphere. 2007,66(6):977~984.
42. De Ruyck H, De Ridder H. Determination of tetracycline antibiotics in cow's milk by liquid chromatography/tandem mass spectrometry.Rapid Commun Mass Spectrom. 2007, 21(9):1511~1520.
43. De Wasch K, Okerman L, Croubels S.Detection of residues of tetracycline antibiotics in pork and chicken meat: correlation between results of screening and confirmatory tests [J].The Analyst, 1998, 123(12): 2737~2741.
44. Fujii A. Pharmacological effect of royal jelly [J]. Honeybee Science, 1995 ,16(1) :97 ~104.
45. Fujiwara S, Imai J, Yaeshima T, et al. A potent antibacterial protein in royal jelly. Purification and determination of the primary structure of royalisin[ J ] . J Biol Chem, 1990, 265(19):11333~11337.
46. Furusawa R. Liquid chromatographic determination of oxytetracycline in milk.J ChromatogrA, 1999, 839 (2): 247~251.
47. Gilliam, M, Argauer R. J. Oxytetracycline residues in surplus honey, brood nest honey and larvae after medication of colonies of honey bees, Apis mellifera, with antibiotic extender patties, sugar dusts, and syrup sprays. Enviro. Entomol 1981,10: 479~482.
48. Giulio S. Determination of sugars in royal jelly by HPLC. Apidologie 2006, 37: 84~90.
49. Gratacós-Cubarsí M, Fernandez-García A, Picouet P, Valero-Pamplona A, García-Regueiro JA, Castellari M. Formation of tetracycline degradation products in chicken and pig meat under different thermal processing conditions. J Agric Food Chem, 2007, 55(11):4610~4616.
50. Heggie, William, Galindro , eta1.Process for preparation of 4 - (des - dimethylamino) - tetracyclines. US.Patent 2000, 6143161.
51. Hisao O, Yoshitomo I, Norihisa K, Keiichi U, Masuo Y, Ken-Ichi H, Makoto S. Improvement of chemical analysis of antibiotics XII. Simultaneous analysis of seven tetracyclines in honey. J ChromatogrA 1987, 400: 253～ 261.
52. Hisao O, Yuko I, Hiroshi M. Chromatographic analysis of tetracycline antibiotics in foods. J ChromatogrA 2000, 393: 285-296.
53. Kamakura M, Mitant N, Fukuda T, etal. Antifatigue effect offresh roual jelly in mice. J Nutr Sci Vitaminol (Tokyo), 2001,47(6) :394 ~401.
54. Lee H J,Le M H, Ryu P D,et a1.Enzyme- linked immunosorbent assay for screening the plasma residues of tetracycline antibiotics in pigs [R].The Journal of veterinary medical science/the Japanese Society of Veterinary Science, 2001.63(5): 553~556.
55. LongAR, Hsieh LC, Malbrough MS, et al. Matrix solid - phase dispersion (MSPD) isolation and liquid chromatographic determination of oxytetracycline, tetracycline, and chlortetracycline in milk [ J ] 1J Assoc Off Anal Chem, 1990, 73 (3) : 379 ~ 384.
56. Marc C, Mario S, Siska C, et a1.Quantitative multi -residue analysis of tetracyclines and their 4- epimers in pig tissues by high - performance liquid chromatography combined with positive- ion electro spray ionization mass spectrometry [J].Analytica Chimica Acta.2003, 492 (1 - 2): 199~213.
57. Mascher A, LavagnoLi S, CuratoLo M . Determination of residual oxytetracycline in honey with an immunoassay kit. ApidoLogie,1996, 27: 229~233.
58. Matsui T, Yukiyoshi A, Doi S , et al . Gastrointestinal enzyme production of bioactive peptides from royal jelly protein and their antihypertensive ability in SHR[J ] . J Nultr Bioch 2002,13 (2) : 80 ~ 86.
59. Nagai T, Inoue, R. Preparation and the functional properties of water extract and alkaline extract of royal jelly. Food Chemistry 2004, 84, 181~186.
60. Nagai T, Sakai M, Inoue R. et al. Antioxidative activities of some commercially honeys, royal jelly and propolis [J]. Food Chemistry, 2001,75(2) :237~240.
61. Naoto Furusawa. An organic solvent - free method for determining oxytetracycline in COW ′ S milk [J].Solutions for separation scientists- LC/GC asia pacific, 2003, 6(3): 23～ 28.
62. Okamoto I, Taniguchi Y, Kunikata T, et al. Major royal jelly protein 3 modulates immune responses in vitro and in vivo[J]. Life Science 2003 ,73(16) :2029~2045.
63. Reybroeck W, Ooghe S, De Brabander H, Daeseleire E. Validation of the tetrasensor honey test kit for the screening of tetracyclines in honey J Agric Food Chem, 2007, 55(21):8359~8366.
64. Samanidou VF, Nikolaidou KI, Papadoyannis IN. Development and validation of an HPLC confirmatory method for the determination of seven tetracycline antibiotics residues in milk according to the European Union Decision 2002/657/EC.J Sep Sci. 2007 (15):2430~2439.
65. Santiago Valverde R, Sánchez Pérez I, Franceschelli F, Martínez Galera M, Gil García MD. Determination of photoirradiated tetracyclines in water by high-performance liquid chromatography with chemiluminescence detection based reaction of rhodamine B with cerium (IV).J Chromatogr A, 2007,1167(1):85~94.
66. Scheer M. Studies on the antimicrobial agents against five pathogens isolated in calves in the Netherlands [J].VetQuart.1900.12( 4) : 212～ 219.
67. Schneider MJ, Braden SE, Reyes-Herrera I, Donoghue DJ.Simultaneous determination of fluoroquinolones and tetracyclines in chicken muscle using HPLC with fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2007,846(1-2):8~13.
68. Schneider MJ, Darwish AM, Freeman DW. Simultaneous multiresidue determination of tetracyclines and fluoroquinolones in catfish muscle using high performance liquid chromatography with fluorescence detection.Anal Chim Acta, 2007,586(1-2):269~274.
69. Smyrniotakis CG, Archontaki HA. C18columns for the simultaneousdetermination of oxytetracycline and its related substances by reversed-phase high performance liquid chromatography and UV detection.J Pharm Biomed Anal. 2007, 43(2):506~514.
70. Spisso BF, de Oliveira e Jesus AL, de Araújo MA Jr, Monteiro MA.Validation of a high-performance liquid chromatographic method with fluorescence detection for the simultaneous determination of tetracyclines residues in bovine milk.Anal Chim Acta, 2007, 581(1):108~117.
71. Stead SL, Caldow M, Sharma A, Ashwin HM, Sharman M, de-Rijk A, Stark J. New method for the rapid identification of tetracycline residues in foods of animal origin - using the PremiTest in combination with a metal ion chelation assay.Food Addit Contam,2007, J24(6):583-589.
72. Sun XY, Chen H, Gao H, Guo XQ. Screening of tetracycline residues in fish muscles by CCD camera-based solid-surface fluorescence. J Agric Food Chem, 2006, 54(26):9687~9695.
73. Sver L, Orsolic N, Tadic Z, et al. A royal jelly as a new potential immunomodulator in rats and mice[J] . Comp Immun Microbiol Infect Dis , 1996 ,19(1) :31~38.
74. Tamura T. Royal jelly from the standpoint of clinical pharmacology[J] . Honeybee Science 1985, 6(1) :117~124.
75. Traviesa-Alvarez JM, Costa-Fernández JM, Pereiro R, Sanz-Medel A. Direct screening of tetracyclines in water and bovine milk using room temperature phosphorescence detection.Anal Chim Acta, 2007,589(1):51~58.
76. Ueno R, Sangrungruang K, Miyakawa M.A simplifiedmethod for the determination of several fish drugs in edible fish and shrimp by high - performance liquid chromatography [J].Food Research Intemationa1.1999,32 (9):629～ 633.
77. Vega D, Agüí L, González-Cortés A, Yá?ez-Sede?o P, Pingarrón JM. Voltammetry and amperometric detection of tetracyclines at multi-wall carbon nanotube modified electrodes.Anal Bioanal Chem,2007 ,389(3):951~858.
78. Wang G Y, Lin ZB. Effects of 102hydroxy222decenoie acid on phagocytosis and cyktokines production of peritoneal macrophages in vitro [J ] . Acta Pharmacol Sin, 1997, 18 (2):180 ~182.
79. White CR. Optimization of a liquid chromatographic method for determination of oxytetracycline, tetracycline, and chlortetracycline in milk.J Assoc of Anal Chem Int, 1993, 76 (3): 549 ~554.
80. Ye Z, Weinberg HS, Meyer MT. Trace analysis of trimethoprim andsulfonamide, macrolide, quinolone, and tetracycline antibiotics in chlorinated drinking water using liquid chromatography electrospray tandem mass spectrometry.Anal Chem. 2007, 79(3):1135~1144.
81. Yoshitomo I, Hisao O, Norihisa K, Masuo Y, Ken-Ichi H, Makoto S. Improvement of chemical analysis of antibiotics : XIII. Systematic simultaneous analysis of residual tetracyclines in animal tissues using thin-layer and high-performance liquid chromatography J ChromatogrA 1987, 411: 313～ 323.