Bi Yun-Tian, Kang Yao-Ren, Woshur Ghulam, Ding Hao-Zhe, Wang Shan-Shan, Qiu Xiang-Jun
School of Basic Medical Sciences, Henan University of Science and Technology, Luoyang 471023, China.
Evid Based Complement Alternat Med. 2022 Aug 17;2022:2350560. doi: 10.1155/2022/2350560. eCollection 2022.
The effect of Chaihu Shugan pills (CHSG) on the pharmacokinetics of duloxetine and its metabolite 4-hydroxyduloxetine in beagle dogs was investigated by establishing an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to simultaneously measure the concentrations of duloxetine and 4-hydroxyduloxetine in beagle dog plasma. Duloxetine and 4-hydroxyduloxetine were separated on the UPLC-C18 column after acetonitrile precipitation and detected by mass spectrometry with multireaction detection mode (MRM). Six adult healthy beagle dogs (weighing 7-9 kg, male and female) were randomly selected and examined for a single-dose administration of duloxetine hydrochloride (2 mg/kg, control group) and oral administration of CHSG (0.3 g/kg) twice daily for 15 consecutive days followed by a single-dose administration of duloxetine hydrochloride (2 mg/kg, experimental group) using the self-control method. All plasma samples were treated in the same way, and then the concentrations of duloxetine and 4-hydroxyduloxetine were determined using the established UPLC-MS/MS method. The obtained data were subjected to DAS 2.0 software to calculate the pharmacokinetic parameters, and SPSS 20.0 software was used to compare the differences between the two groups. Duloxetine and 4-hydroxyduloxetine had a good linear relationship in the ranges of 1-1000 ng/ml and 0.1-100 ng/ml, and the lower limits of quantification (LLOQ) were 1 ng/mL and 0.1 ng/ml, respectively. The precision, accuracy, extraction recovery, matrix effect, and stability meet the requirements of the guiding principles. After combination with CHSG, and AUC of duloxetine decreased by 49.33% and 13.08%, respectively, and was shortened to 10.17 h; and AUC of 4-hydroxyduloxetine decreased by 71.47% and 48.78%, respectively, and was shortened to 7.97 h. The UPLC-MS/MS method was fully developed to simultaneously measure the plasma concentration of duloxetine and its metabolite 4-hydroxyduloxetine in beagle dogs. CHSG could slow down the absorption of duloxetine, induce the metabolism of duloxetine and 4-hydroxyduloxetine in beagle dogs, and reduce plasma exposure.
通过建立超高效液相色谱-串联质谱法（UPLC-MS/MS）同时测定比格犬血浆中度洛西汀及其代谢物4-羟基度洛西汀的浓度，研究柴胡疏肝丸（CHSG）对度洛西汀及其代谢物在比格犬体内药代动力学的影响。度洛西汀和4-羟基度洛西汀经乙腈沉淀后在UPLC-C18柱上分离，采用多反应监测模式（MRM）进行质谱检测。随机选取6只成年健康比格犬（体重7-9 kg，雌雄不限），采用自身对照法，单次给予盐酸度洛西汀（2 mg/kg，对照组），并连续15天每天口服CHSG（0.3 g/kg）两次，之后单次给予盐酸度洛西汀（2 mg/kg，实验组）。所有血浆样本均采用相同方法处理，然后使用建立的UPLC-MS/MS法测定度洛西汀和4-羟基度洛西汀的浓度。将所得数据用DAS 2.0软件计算药代动力学参数，并用SPSS 20.0软件比较两组间差异。度洛西汀和4-羟基度洛西汀在1-1000 ng/ml和0.1-100 ng/ml范围内线性关系良好，定量下限（LLOQ）分别为1 ng/mL和0.1 ng/ml。精密度、准确度、提取回收率、基质效应和稳定性均符合指导原则要求。与CHSG合用时，度洛西汀的(t_{max})和AUC分别降低49.33%和13.08%，(t_{max})缩短至10.17 h；4-羟基度洛西汀的(t_{max})和AUC分别降低71.47%和48.78%，(t_{max})缩短至7.97 h。已充分建立UPLC-MS/MS法同时测定比格犬血浆中度洛西汀及其代谢物4-羟基度洛西汀的浓度。CHSG可减缓度洛西汀的吸收，诱导比格犬体内度洛西汀和4-羟基度洛西汀的代谢，并降低血浆暴露量。
