B vitamins are essential water-soluble micronutrients for humans, and meat is an important dietary source. However, the complex matrix of meat samples (high protein and high fat) leads to poor selectivity and severe matrix interference in traditional analytical methods. To meet the demand for simultaneous determination of multiple B vitamins in complex meat matrices, an analytical method based on solid-phase extraction (SPE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was established for the simultaneous determination of 11 B vitamins in pork, beef, and fish. Samples were subjected to enzymatic hydrolysis (compound enzymes, pH 5.5, 37°C, 1.5 h) followed by buffer extraction. The extracts were purified using a Captiva EMR-Lipid SPE cartridge. Chromatographic separation was achieved on an Agilent HILIC Plus column (2.1 × 100 mm, 3.5 μm) using 0.2% (V/V) formic acid-10 mmol·L-1 ammonium acetate solution and acetonitrile as the mobile phase. Detection was performed under electrospray ionization in positive mode with multiple reaction monitoring (MRM), and quantification was conducted using an external standard method. The results showed that all 11 target analytes exhibited good linearity (R2 > 0.9990). The linear ranges were 5-1000 ng·mL-1 for eight vitamins, 10-1000 ng·mL-1 for pantothenic acid, and 15-1000 ng·mL-1 for folic acid and vitamin B12. The limits of detection (LOD) and limits of quantification (LOQ) were 0.1-2.8 μg·kg-1 and 0.5-9.8 μg·kg-1, respectively. The average recoveries ranged from 85.72% to 107.17% at three spiking levels in pork, beef, and fish matrices, with relative standard deviations (RSDs) of 1.13%-6.56%. The matrix effects ranged from 2.38% to 18.47%. All performance parameters met the quantitative requirements of GB/T 27404-2026. This method provides theoretical and technical support for the nutritional evaluation and quality control of meat products.