羅丹明B和羅丹明6G皆屬於Xanthene Dyes，除了被當作顏色添加劑而廣泛的使用在化妝品、食品、藥物之中，也被當作著色劑而常被使用在紡織和塑膠工業上。雖然這類染料被大量使用在不同的領域之中，但已經有許多研究指出這些染料會對皮膚、眼睛和呼吸道造成刺激和傷害，而且Xanthene Dyes的致癌性、生殖上和發育上的相關毒性皆已經被證實。
　　本研究利用高效液相層析儀搭配螢光偵檢器(High Performance Liquid Chromatography-Fluorescence Detector，簡稱HPLC-FLD)發展出一套簡易的方法來檢測台灣環境水樣中羅丹明B和羅丹明6G的含量。最佳的分離條件為使用Waters Atlantis? T3-C18 (150 mm * 2.1 mm i.d., 3 μm)當作分離管柱，以0.1 % H3PO4水溶液：乙?(CH3CN)＝55 %：45 %當作動相，在0.2 mL/min的流速下使用等位沖提(Isocratic Elution)進行分離。為了使樣品可達到最佳的萃取效果，本研究中也嘗試在固相萃取法的條件上進行探討，如pH值、管柱材質、沖提液等，而最佳的萃取條件為在pH2-3，使用HLB當作固相萃取管柱並利用4 %甲酸於甲醇溶液中進行沖提。本研究中羅丹明B和羅丹明6G的定量極限分別為0.5 ng/L和2 ng/L，其環境水樣中的萃取回收率在55%至90 %之間，相對標準偏差在5.5 %至17 %之間，待測物於環境水樣中的含量則在0.7 ng/L至81 ng/L之間。

Rhodamine B (RB) and rhodamine 6G (R6G) belong to the xanthene dyes, which are widely used as color additive in cosmetic, food, pharmaceutical and also used as colorant in textiles, plastic industry, etc. Although there are widely used in variety domain, many research pointed out that these compound
would cause irritation to the skin, eyes and respiratory tract, moreover, the carcinogenicity, reproductive and developmental toxicity of xanthene dyes have also been reported.
　　A simple method was developed to detect two xanthene-type dyes, rhodamine B (RB) and rhodamine 6G (R6G) in Taiwan’s environmental water samples by high performance liquid chromatography coupled with fluorescence detector (HPLC-FLD). The contents of these two xanthene-type dyes in environmental water sample were extracted by solid-phase extraction (SPE). The optimization separation condition was Atlantis? T3-C18 (150 mm * 2.1 mm i.d., 3 μm) as separation column, and 0.1% H3PO4 solution : CH3CN=55% : 45% as mobile phase, flow rate is 0.2 mL/min for isocratic elution. The SPE condition such as various pH, SPE cartridges, and eluents were also evaluated, the optimization extraction condition was that using HLB cartridge, elute with 4 % HCOOH in MeOH at pH2-3. Experimental results indicate that the proposed method is precise and sensitive in analyzing rhodamine B and rhodamine 6G, and the limits of quantitation were 0.5 ng/L for R6G and 2 ng/L for RB. The recovery of analytes in spiked environmental water samples ranged from 55 % to 90 %, and the precision (RSD) ranged from 5.5 % to 17 %. The concentration of analytes were detected in environmental water samples ranging from 0.7 ng/L to 81 ng/L.

第六章　參考文獻
 王傳現，韓麗，方曉明，李波，王敏，倪昕路：食品中羅丹明B的高效液相色譜螢光檢測，儀器分析，第27-30頁，2008第一期。
 邱永亮，魏盛德編譯：科學圖書大庫＂染色化學(合)”，第182-188 頁，民國89 年。
 游碧堉，蔡三福，呂水淵，張敬宜，李彥芸，陳敏貞：農藥對溫血動物毒性測試技術之研究，行政院農業委員會農業藥物毒物試驗所，國科會審議編號：9621012104140202P145，民國96年。
 Behrens, H.; Beims, U.; Dieter, H.; Dietze, G.; Eikmann, T.; Grummt, T.; Hanisch, H.; Henseling, H.; K??, W.; Kerndorff, H.; Leibundgut, C.; M?ller-Wegener, U.; ?nnefahrt, I.; Scharenberg, B., Toxicological and ecotoxicological assessment of water tracers, Hydrogeology J., 2001, 9, 321–325.
 Desiderio, C.; Marra, C.; Fanali, S., Quantitative analysis of synthetic dyes in lipstick by micellar electrokinetic capillary chromatography, Electrophoresis, 1998, 19, 1478-1483.
 Dire, D.; Wikinson, J.A., Acute exposure to rhodamine B, J. Toxicol Clin Toxicol, 1987, 25, 603-607.
 Franke, C.; Westerholm, H.; Niessner, R., Solid-phase extraction of the fluorescence tracers uranine and sulphorhodamine B, Water Res., 1997, 31, 2633-2637.
 French J. E., Toxicology and carcinogenesis studies of rhodamine 6G in F344/N rats and B6C3F1 mice, National Toxicolgy Program Technical Report Series NO. 364, 1989.
 Gagliardi, L.; De Orsi, D.; Gavazzutti, G.; Multari, G.; Tonelli, D., HPLC Determination of rhodamine B (C.I. 45170) in Cosmetic Products, Chromatographia, 1996, 43, 76-78.
 Harris, D.C., Quantitative Chemical Analysis 6th ed., W.H. Freeman and Company, 2003.
 Hofstraat, J. W.; Steendijk, M.; Vriezekolk, G.; Schreurs, W.; Broer, G. J. A. A.; Wijnstok, N., Determination of rhodamine WT in surface water by solid-phase extraction and HPLC with fluorescence detection, Water Res., 1991, 25, 883-890.
 IARC, monographs on the Evaluation of Carcinogenic Risk to Humans, 1978, 16, 221.
 Kaji, T.; Kawashima, T.; Yamamoto, C.; Sakamoto, M., Rhodamine B inhibits collagen synthesis by human lip fibroblasts in culture, Toxicol Lett, 1992, 61, 81-87.
 Lampidis, T.J.; Salet, C.; Moreno, G.; Chen, L.B., Comparative effects of the mitochondrial probe Rhodamine 123 and related analogs on the function and viability of pulsating myocardial cells in culture, Agents Actions, 1984, 14, 751.
 Laane, R.W.P.M.; Manuels M.W.; Staal W., A procedure for enriching and cleaning up rhodamine B and rhodamine WT in natural water, using Sep-pak C18 cartridge. Wat. Res. 1984, 18, 163-165
 Lee, C.K.; Liu, S.S.; Juang, L.C.; Wang, C.C.; Lin, K.S.; Lyu, M.D., Application of MCM-41 for dyes removal from wastewater, J. Hazard. Mater., 2007, 147, 997-1005.
 Lewis, I.L.; Patterson, R.M.; McBay, H.C., The effects of Rhodamine B on the chromosomes of Muntiacus muntjac, Mutat Res., 1981, 88, 211-216.
 Mitrowska, K.; Posyniak, A.; Zmudzki, J., Determination of malachite green and leucomalachite green in carp muscle by liquid chromatography with visible and fluorescence detection, J. Chromatogr. A., 2005, 1089, 187-192.
 Naga, S.; Koike, H., Separation of Food Dyes by High Performance Liquid Chromatography Using Methylguanidine as ion-pair Reagent, Anal. Sci., 1990, 6, 473-474.
 Nestmann, E.R.; Douglas, G.R.; Matula, T.I.; Grant, C.E.; Kowbel, D.J., Mutagenic activity of rhodamine dyes and their impurities as detected by mutation induction in Salmonella and DNA damage in Chinese hamster ovary cells, Cancer Res., 1979, 39, 4412-4417.
 Parodi, S.; Taningher, M.; Russo, P.; Pala, M.; Tamaro, M.; Monti-Bragadin, C., DNA-damaging activity in vivo and bacterial mutagenicity of sixteen aromatic amines and azo-derivatives, as related quantitatively to their carcinogenicity. Carcinogenesis, 1981, 2, 1317-1326.
 Pourreza, N.; Rastegarzadeh, S.; Larki, A., Micelle-mediated cloud point extraction and spectrophotometric determination of rhodamine B using Triton X-100, Talanta., 2008, 77, 733-736.
 Rochat, J.; Alary, J.; Molinari, J.; Charriere R., Srparations physicochimiques de colorants xanthrniques utilisrs comme traceurs en hydrologic, J. Hydrol., 1975, 26, 277-293
 RTECS CCINFO online - May 2005.
 Sachdeva, S.; Kumar, A., Preparation of nanoporous composite carbon membrane for separation of rhdoamine B dye, J. Membr. Sci., 2007, 329, 2-10.
 Scalia, S.; Simeoni, S., Assay of Xanthene Dyes in Lipsticks by Inverse Supercritical Fluid Extraction and HPLC, Chromatographia, 2001, 53, 490-494.
 Skoog, D.A.; Holler, F.J.: Nieman, T.A., Principles of instrumental analysis, 5th ed., Thomson Learing, 1998.
 Sun, H.; Wang, F.; Ai, L., Determination of banned 10 azo-dyes in hot chili product by gel permeation chromatography-liquid chromatography-electrospray ionization-tandem mass spectrometry, J. Chromatogr. A., 2007, 1164, 120-128.
 Tarbin, J.A.; Chan, D.; Stubbings, G.; Sharman. A., Multiresidue determination of triarylmethane and phenothiazine dyes in fish tissues by LC-MS/MS, Analytica Chimica Acta., 2008, 625, 188-194.
 The Star, 30 May 2007(Website: http://www.thestar.com.my) Carcinogenic rhodamine B found in Malaysian foodstuffs, Focus on pigments, 2007, 2007, Page 7.
 Van Bommel, M.R.; Berghe, I.; Wallert, A.M.; Boitelle, R.; Wouter, J., High-performance liquid chromatography and non-destructive three-dimensional fluorescence analysis of early synthetic dyes, J. Chromatogr. A., 2007, 1157, 260-272.
 Wang, C.C.; Masi, A.N.; Fern´andez, L., On-line micellar-enhanced spectrofluorimetric determination of rhodamine dye in cosmetics, talanta., 2008, 75, 135-140.
 Wu, X.; Zhang, G.; Wu, Y.; Hou, X.; Yuan, Z., Simultaneous determination of malacie green, gentian violet and their leuco-metabolites in aquatic products by high-performance liquid chromatography-linear ion trap mass spectrometry, J. Chromatogr. A., 2007, 1172, 121-126.