Abstract: The multi-component pharmacokinetic of traditional Chinese medicine (TCM) is vital to clarify the theory and mechanism of TCM. Based on information of absorption, distribution, metabolism, and excretion dynamic changes for active components of TCM in vivo, the material basis for the safety and efficacy can be elucidated via combination with pharmacodynamics. However, due to the complexity and diversity of TCM components, and their complicated and unclear interactions with biological system, there are many bottleneck problems for the multi-component pharmacokinetics of TCM, such as the numerous chemical composition and diverse metabolic reaction make it difficult to identify their structures and interpret metabolic pathway in vivo; and unclear active components, low concentrations of target components in biological samples, endogenous interferences from matrix and lack of authentic standards bring difficulties to establish qualitative and quantitative methods. In recent years, the pretreatment methods for biological samples have been well developed. The separation and extraction efficiency for multi-component of TCM in vivo and in vitro have been significantly improved using various high-throughput, fast and automated pretreatment techniques. The development of chromatography and mass spectrometry promotes the rapid comprehensive and accurate discovery and quantification for multi-component of TCM. Novel analytical methods such as mass spectrometry imaging were used to provide new strategies for multi-component pharmacokinetics of TCM. In this paper, the development and application of analytical techniques for multi-component pharmacokinetic of TCM in the past 10 years were summarized, and a brief perspective was discussed. Firstly, the pretreatment of biological sample was introduced, the advantages and disadvantages of some usual sample pretreatment methods were compared such as protein precipitation and liquid-liquid extraction, solid phase extraction, microextraction, microdialysis. The applications of these different pretreatment techniques in the pharmacokinetics research of TCM were interviewed. Then, the principles and developments of different chromatographic techniques including high performance liquid chromatography and gas chromatography used in multi-component pharmacokinetic of TCM were introduced. Finally, the mass spectrometry ionization techniques were summarized and discussed including electrospray ionization and atmospheric pressure chemical ionization for liquid chromatography, electron impact and chemical ionization for gas chromatography, desorption electrospray ionization, direct analysis in real time, mass spectrometry imaging.