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Phyto-pharmacokinetics and Environmental Residues of Tetracycline and Sulfamethoxazole in Chinese Cabbages (Brassica rapa Chinensis) and Water Spanish (Ipomoea aquatica)
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|摘要:||四環黴素類和磺胺劑是廣泛使用於飼料添加和動物臨床治療之抗生素，可在糞肥、土壤和水中經常檢出ppm到ppb等級的殘留。以往的研究大部分著重於抗生素在植物的累積情形，甚少以藥物動力學的觀點來探討藥物進入植物體內的命運，包括藥物的吸收、分布、代謝及排除等過程。本研究就畜牧業使用量較多的四環黴素(TC)及環境中穩定性較高的磺胺甲基噁唑(SMX)，以台灣常食用的小白菜和空心菜為主題，透過液相層析串聯質譜儀檢測兩菜各組織的抗生素濃度。將蔬菜種植在100 µg mL-1 TC或SMX的水耕環境下24小時並觀察其藥物動力學行為，結果顯示，兩藥在兩菜的藥物動力學模式有明顯的差異，TC和SMX可在很短的時間內在蔬菜內累積，每克小白菜可吸收TC達160 µg，SMX達18 µg，而每克空心菜可吸收TC達77 µg，SMX達38 µg。TC在小白菜根的累積可達水耕液濃度21倍，在葉的累積也達1.2倍，而TC和SMX在空心菜各組織並無生物累積現象。排除試驗顯示TC和SMX的主要排除發生在試驗的前30分鐘，24小時在小白菜可排除11%(TC)和8%(SMX)的菜中總量，而在空心菜可排除25%(TC)和28%(SMX)。以非隔室模型藥物動力學分析，TC在小白菜根的Cmax(最高濃度)達1124 µg mL-1，是SMX在小白菜的34倍，亦是TC和SMX在空心菜根的32倍和182倍；SMX在兩菜的分布容積(Vd)比TC高出3-6倍，SMX在小白菜根的半衰期(88小時)最長，而TC半衰期(22小時)最短，而TC和SMX在空心菜根的半衰期則介於之間，TC在小白菜根的AUC(曲線下面積)最高，是TC在空心菜和TC與SMX在空心菜根的數倍之多，TC在小白菜根的平均滯留時間比SMX小，但在空心菜的情形則相反，綜合以上，兩藥在兩菜的藥物動力學表現有明顯的差異。將小白菜和空心菜種植在含藥的土壤21天後，兩菜在各組織間所測得的藥物濃度皆低於10 μg mL-1，顯示在土耕環境下兩菜對於TC和SMX的吸收並不顯著，原因可能和藥物在土中受土壤酸鹼值、離子強度和濕度等特性而影響吸收有關。而從台灣各地區豬場周邊及傳統市場植物的抗生素殘留調查(蔬菜樣本數265，雜草樣本數70，總樣本數共計355株)，28%蔬菜和64%雜草為抗生素陽性且至少有一種以上的抗生素殘留，豬場周邊蔬菜的抗生素陽性率是傳統市場的兩倍，殘留的抗生素中以羥四環素(20%)和金黴素(33%)的陽性率較高，而SMX陽性率皆低。所測得蔬菜的殘留濃度以小白菜的去氧羥四環黴素最高(231 ppb)，雜草中以香附子的羥四環黴素濃度(746 ppb)最高，而豬場排放水和周邊土壤的抗生素陽性率為100%和87%。本研究是第一個以蔬菜為主題，以藥物動力學模型分析動物常用抗生素在植物內移動的研究，顯示有特別意義和探討價值。|
Tetracyclines (TCs) and sulfonamides (SAs) are common applications as food-additives and therapeutic treatment in veterinary medicine. In recent years, antibiotic residues are increasingly being monitored in slurry, soils and surface waters as ground waters have been reported to have concentrations ranging from micrograms to milligrams per liter in many countries. To the best of our knowledge, very few studies have examined the fate of antibiotics in the edible plants other than to study their accumulations and none of them was studied from the pharmacokinetic perspectives, which includes the adsorption, distribution, metabolism and elimination. Therefore, the main purposes of the study were focused on two popular vegetables in Taiwan and to demonstrate the novel use of pharmacokinetic approaches to study the movements of antibiotics in edible plants. Brassica rapa Chinensis and Ipomoea aquatic were grown in cultivation fluids containing 100 µg mL-1 TC or SMX for 24 hours and drug concentrations were analyzed by HPLC with LC-MS-MS detection. The absorption results indicated that ephemeral vegetables could have high capacity accumulating antibiotics within hours (up to 160 µg g-1 for TC and 18 µg g-1 for SMX in B. rapa Chinensis and 77 µg g-1 for TC and 38 µg g-1 for SMX in I. aquatica). TC concentration in the root of B. rapa Chinensis (Cmax) could reach 21 times higher than that in the cultivation fluid and concentration in the leave reach 1.2 times higher than in the cultivation fluid, but there were no bioaccumulation for TC and SMX in I. aquatica . For elimination, B. rapa Chinensis could release 11% (TC) and 8% (SMX) of the drugs back into the cultivation fluid while in I. aquatica they were released 25% (TC) and 28% (SMX) for 24 hours. Non-compartmental analysis indicated that TC and SMX exhibited distinct PK behaviors in vegetables grown hydroponically. Cmax of TC could reach as high as 1124 µg mL-1 which was over 34 folds more than SMX concentration in the roots of B. rapa Chinensis and more than 32 folds and 182 folds of TC and SMX in the root of I. aquati. Based on the volume of distribution (Vd), SMX was 3-6 times more extensively distributed than TC. Both antibiotics showed slow albeit evident elimination phases with elimination half-lives ranging from 22 to 88 hours. When vegetables were exposed to 100 μg mL-1 TC or SMX in soil for 21 days, the detectable levels of either drug were very low in both vegetables (below 10 μg mL-1). Possible reasons are likely associated with the degradation of drugs and the complexity of rhizosphere (soil characters, microorganism, pH, ion strengths, and humidity) that hindered the drug absorption. In the field investigations of a total of 335 samples (vegetables 265 and weeds 70) on the levels of TCs and SMX near pig farms or from traditional market in Taiwan, results showed that 28% of vegetables and 64% of weeds near pig farms contained at least one TCs. The detection rate of vegetables was twice higher near the pig farm than in the traditional market. OTC(20%) and CTC(33%) were most commonly detected while the detection rate was very low in SMX. DC level could reach as high as 231 ng g-1 in Chinese cabbage and OTC level 746 ng g-1 in nut grass among all plants. The soils and the discharges water near the pig farms also showed very high contamination rate (87% and 100%) of these two drugs. We have demonstrated for the first time that pharmacokinetics approaches commonly used in animal model could be used to at least partially describe drug behaviors in the plants. Phyto-pharmacokinetics could be a new area worth development of new models for risk assessment of veterinary drugs.
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