Inulin-type fructooligosaccharides (FOS), which are a group of compounds recognized as natural food ingredients and classified as dietary fiber in almost all European countries, are formed by β-(2→1) linked fructofuranosyl unit on the end of sucrose molecule. In this study, the methods for extraction, separation and purification of FOS were investigated, and three analytical methods for the determination of FOS in plants were developed respectively. The major achievements of this study are summarized as follows: (1) A fast protein liquid chromatography coupled with refractive index detection (FPLC-RID) method was firstly developed for separation and purification of FOS with different DP (DP3 to DP13) from burdock. Total FOS were purified on Bio-Gel P columns eluted with water at the flow rate of 0.3 ml/min, and the HPLC-ELSD method was used for determination of the purities of FOS. The separated FOS were identified by their methylation analysis, MS and NMR data. (2) A twice development HPTLC method was established for simultaneously qualitative and quantitative analysis of seven FOS (DP3~9) in Morinda officinalis and Arctium lappa. The analysis was performed on silica gel 60 plate with n-butanol - isopropanol - water - acetic acid (7:5:2:1, v/v/v/v). The bands were revealed by reaction with 1-naphthol-sulfuric acid reagent and densitometrically determined at 585 nm wavelength. (3) An HPLC-ELSD and microwave-assisted extraction method were developed for determination of seven FOS (DP3~9), as well as fructose, glucose and sucrose in Morinda officinalis and Arctium lappa from different regions. The separation was performed on a column B (4.6×250 mm id, 3.5 μm) with gradient iv elution. (4) An HPLC-CAD method was developed for simultaneous determination of FOS (DP 3 ~13) in different samples from Liliaceae, Asteraceae, Campanulaceae, and Rubiaceae families. LC-MS/MS analysis was also carried out for qualitative analysis of FOS in these samples. In summary, An FPLC-RID method was developed for fast preparation and purification of FOS (DP3-DP13) for the first time. Three analytical methods (HPTLC, HPLC-ELSD, HPLC-CAD) for quantitative analysis of FOS in different samples were also developed, which provided powerful tools for quality control of these plants.