浸泡条件下孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾 组织中分布及消除规律研究

TISSUE DISTRIBUTION AND ELIMINATION OF MALACHITE GREEN AND ITS METABOLITE LEUCOMALACHITE GREEN FROM CHANNEL CATFISH (IETALURUS PUNETAUS) AFTER BATH TREATMENT

  • 摘要: 以7 mg/L的孔雀石绿浸泡斑点叉尾 苗种5min后将其饲养于池塘的网箱中, 研究了在养殖模式下孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾 苗种各组织中的分布及消除规律。采用高效液相色谱串联质谱法(HPLC-MS/MS)分析孔雀石绿及其代谢物隐色孔雀石绿在斑点叉尾 血液、肌肉、皮肤、肝脏、肾脏组织中的浓度水平。采用药代动力学分析软件3p97对血药浓度时间数据进行分析。结果表明, 孔雀石绿和隐色孔雀石绿血药浓度时间曲线符合有吸收二室模型, 动力学方程分别为: C孔雀石绿 =683.063 e-0.248 t+ 11.176 e-0.006 t- 694.239e-0.333 t, C隐色孔雀石绿 =757.240 e-0.222 t + 14.474 e-0.007 t 771.714 e-0.382 t。血液中孔雀石绿和隐色孔雀石绿达峰时间Tpeak分别为3.480和3.623h, 峰浓度值Cmax分别为81.560和159.619 ng/mL, 表观分布容积Vd/F分别为37.689和21.125 L/kg, 分布相的一级速率常数分别为0.248和0.222/h, 消除相的一级速率常数分别为 0.006和0.007/h, 吸收半衰期T(1/2) 分别为2.794和3.124h, 消除半衰期T(1/2)分别为113.068和105.841h, 中央室向周边室转运的一级速率常数K12分别为0.020和0.015/h, 周边室向中央室转运的一级速率常数K21分别为0.159和0.121/h, 药-时曲线下面积AUC分别为2493.944和3601.863 ngh/mL。肌肉、皮肤、肝脏和肾脏组织中孔雀石绿和隐色孔雀石绿浓度水平的结果表明, 孔雀石绿在斑点叉尾 4种组织中浓度由高到低的顺序是皮肤肌肉肾脏肝脏, 其中斑点叉尾 皮肤组织易蓄积孔雀石绿, 其残留时间最长, 肝脏组织由于对孔雀石绿有极强的代谢转化功能而浓度较低。孔雀石绿在肌肉、皮肤、肝脏和肾脏组织中的消除方程分别为C=5.570 e-0.009t、C=6.302 e-0.007t、C=4.791 e-0.006t和C=4.591 e-0.002t, 相关系数r20.773, 消除半衰期T1/2肌肉、皮肤、肝脏和肾脏分别为3.2、4.1、4.8和14.4d。肌肉、皮肤、肝脏和肾脏组织中孔雀石绿分别在45、60、30和60d才未被检测到; 隐色孔雀石绿在斑点叉尾 4种组织中浓度由高到低的顺序是肝脏皮肤肌肉肾脏, 残留时间最长的组织也是皮肤组织。隐色孔雀石绿在肌肉、皮肤、肝脏和肾脏组织中的消除方程分别为C=6.491 e-0.004t、C=6.958 e-0.003t、C=6.722 e-0.007t和C=6.162 e-0.002t, 相关系数r20.673, 消除半衰期T1/2肌肉、皮肤、肝脏和肾脏分别为7.2、9.6、4.1和14.4d。肌肉、皮肤、肝脏和肾脏组织中隐色孔雀石绿分别在90、90、60和90d才未被检出。试验期间(2011年5月17日至7月15日)平均水温为26.4℃, 孔雀石绿和隐色孔雀石绿90d后在各组织中才未检测到, 因此, 使用7 mg/L孔雀石绿浸泡2龄斑点叉尾 苗种孔雀石绿及其代谢物隐色孔雀石绿至少应经过2376℃d后才能消除。

     

    Abstract: The distribution and depletion rules of MG and its metabolite LMG in channel catfish was investigated after bath treatment of 7 mg/L MG. Channel catfish were raised in the cages in the pool. Blood, muscle, liver, skin and kidney of channel catfish were analyzed by HPLC/MS/MS. The data of drug concentration-time were analyzed using pharmacokinetics software 3p97. The results showed that blood concentration-time data of MG and LMG were both best fitted using a two-compartment model with first absorption. The pharmacokinetic equations were CMG =683.063 e-0.248t+11.176 e-0.006t- 694.239 e-0.333t and CLMG =757.240 e-0.222t +14.474 e-0.007t 771.714 e-0.382t. Peak concentrations of MG and LMG were 81.560 ng/mL and 159.619 ng/mL at 3.480h and 3.623h, respectively. The distribution volume (Vd/F) of MG and LMG were 37.689 L/kg and 21.125 L/kg, respectively. First-order rate constants for MG and LMG distribution phases () were 0.248/h and 0.222/h, respectively, First-order rate constants for MG and LMG elimination phases () were 0.006/h and 0.007/h respectively. The absorption half-life T(1/2) were 2.794h and 3.124h, respectively. The elimination half-life T(1/2) were 113.068h and 105.841h respectively. First-order rate constants for MG and LMG distributions from central compartment to peripheral compartment (K12) were 0.020/h and 0.015/h respectively. First-order rate constants for MG and LMG distributions from peripheral compartment to central compartment (K21) were 0.159/h and 0.121/h, respectively. The areas under the curve (AUC) were 2493.944 ngh/mL and 3601.863 ngh/mL. The results of MG and LMG concentrations in muscle, skin, liver and kidney of channel catfish showed that the orders of 4 kinds of tissues concentration levels of MG from high to low were skin, muscle, kidney and liver. MG was easy to accumulate in skin, and residue time was the longest. Because MG was metabolited by liver fastly, low concentrations of MG were determined in liver. The Elimination equations of MG in muscle, skin, liver and kidney were C=5.570 e-0.009t, C=6.302 e-0.007t, C=4.791 e-0.006t and C=4.591 e-0.002t. Those correlation coefficients (r2) were 0.773 respectively. The elimination half-life (T1/2) of MG in muscle, skin, liver and kidney were 3.2d, 4.1d, 4.8d and 14.4d, respectively. MG was not detected in muscle, skin, liver and kidney of channel catfish at 45d, 60d, 30d and 60d respectively. The orders of 4 kinds of tissues concentration levels of LMG from high to low were liver, skin, muscle kidney. The tissue with the longest residue time of LMG was also skin. The Elimination equations of LMG in muscle, skin, liver and kidney were C=6.491 e-0.004t, C=6.958 e-0.003t, C=6.722 e-0.007t, and C=6.162 e-0.002t, repectively. The correlation coefficients (r2) of them were 0.673. The elimination half-life of MG in muscle, skin, liver and kidney were 7.2d, 9.6d, 4.1d and 14.4d. LMG was not detected in muscle, skin, liver and kidney of channel catfish at 90d, 90d, 60d and 90d, respectively. MG and LMG were not detected at 90d and the average water temperature was 26.4℃ during the experiment from May 17, 2011 to July 15, 2011. Malachite green and its metabolite Leucomalachite green should at least through 2376℃d to eliminate at 7 mg/L malachite green for 2 age of channel catfish.

     

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