中药复方制剂整体复杂成分体内外含量测定的方法学研究

A methodological study on complicated constituents determination of TCM compound preparation in vitro and vivo

目的:探讨吸波面积测定透骨消痛胶囊中整体复杂成分体内外含量方法的科学性与可行性,为复杂体系中药复方制剂中整体药物总固体物成分的含量测定奠定基础。方法:以透骨消痛胶囊为模型,严格按照分析方法的要求,采用吸波面积法,利用紫外分光光度法在200~500nm下对胶囊内容物甲醇溶液进行全波长扫描,利用Origin软件,求算吸波面积,根据吸波面积和总固体整体成分的总浓度成正比的特点,测得胶囊中整体成分的含量;同时,通过大鼠灌服透骨消痛胶囊混悬液,利用吸波面积法测定了大鼠体内血浆中透骨消痛胶囊总固体物不同时间点血药浓度的动态变化,并用3P97软件计算出房室模型及其药动学参数。结果:透骨消痛胶囊体外方法学考察中药物总固体物在2.08~104.00μg/m L范围内线性关系良好,线性回归方程为:AUAWC=1.1531C+1.9239(r=0.9999),平均加样回收率低、中、高分别为103.66%(RSD=2.06%)、104.21%(RSD=2.82%)、108.63%(RSD=2.95%);大鼠体内方法学考察中,吸波面积与血浆中胶囊总固体物总浓度在1.87~93.60μg/m L范围内,线性关系良好,回归方程:ΔAUAWC=0.6913C+1.7294(r=0.9997),提取回收率低、中、高分别为99.52%(RSD=5.44%)、81.18%(RSD=3.01%)、80.45%(RSD=1.26%),相对回收率分别为100.64%(RSD=7.08%)、104.45%(RSD=3.45%)、107.34%(RSD=1.85%);药代动力学测得大鼠体内药物过程为单室模型,整体药物的Cmax为(10.20±0.586)mg/L,T1/2为(1.901±0.049)h,AUC0~24为(12.718±0.422)mg·h^(-1)·L^(-1)。结论:吸波面积法可用于透骨消痛胶囊中整体药物体内外含量的测试,具有较高的精密度和稳定性,重现性和回收率良好,为中药复方制剂整体复杂成分体内外含量测定的方法学研究提供了较好的启示作用。

Objective： The scientificity and feasibility of complicated constituents determination of the Tougu Xiaotong capsules in vivo and vitro by the method of area under the absorbability-wavelength curve were investigated in order to lay the foundation for the content determination of total solids components- Methods： Based on the model of the Tougu Xiaotong capsules, the method of AUAWC was applied for computing absorbing area- The overall compositions in the capsules were measured according to the characteristics that AUAWC has direct ratio relation with the total concentration of the overall composition in the total solids- Meanwhile, after oral administration, the phase changes in plasma concentration of the Tougu Xiaotong capsules were obtained by the method of AUAWC- And the pharmacokinetics parameters and compartment model were calculated by 3P97 software- Results： The methodological study in vitro showed that there was a good linear relationship at the range of 2-08～104-00μg/m L- The linear regression equation was AUAWC=1-1531C＋1-9239（r=0-9999）- The average recovery of low, middle and high concentration were 103-66%（RSD=2-06%）, 104-21%（RSD=2-82%） and 108-63%（RSD=2-95%） respectively- The methodological study in vivo showed that there was a good linear correlation at the range of 1-87～93-60 μg/m L between the AUAWC and the plasma concentration of the Tougu Xiaotong capsules- The linear regression equation was ΔAUAWC =0-6913 C ＋ 1-7294（r=0- 9997）- The extraction recovery of low, middle and high concentration were 99-52%（RSD=5-44%）, 81-18%（RSD=3-01%） and 80-45%（RSD=1-26%） respectively- The method recovery were 100-64%（RSD=7-08%）, 104-45%（RSD= 3-45%） and 107-34%（RSD= 1-85%） respectively- The pharmacokinetics of the Tougu Xiaotong capsules could be described by single compartment model- The main pharmacokinetic parameters of total components were as follows： Cmax of（10-20±0-586）mg/L, T1/2 at（1-901±0-049）h, AUC0～24 of（12-718±0-422） mg·h-1·L-1- Conclusion： The method of AUAWC can be used for the content determination about the overall drug content of the Tougu Xiaotong capsule in vivo and vitro- The method showed higher precision, stability, good repeatability and recovery- The method of AUAWC could provide a good enlightenment for the methodology study on complicated constituents determination of TCM compound preparation in vivo and vitro.