Difference between HPLC and UPLC
- 1. DIFFERENCE BETWEEN HPLC AND UPLC Dr. Vrushali Sachinkumar Tambe Professor (Department of Pharmaceutical Chemistry) PES Modern College of Pharmacy (for Ladies), Pune, India
- 2. • High-performance liquid chromatography (HPLC) is an important liquid chromatography (LC) technique used for the segregation of different components in mixtures. It is also used for the identification and quantification of compounds. • In 2004, Waters launched and trademarked Ultra Performance Liquid Chromatography (UPLC) which is based upon small, porous particles (sub 2micron particles). This technique achieved dramatic increase in resolution, speed and sensitivity in LC.
- 3. Sr. No. UPLC HPLC 1 Particle size of stationary phase is less than 2 micron (typically 1.7 micron) Particle size of stationary phase is between 3 to 5 micron 2 Operates at higher pressure due to reduced particle size Maximum backpressure of 1000 bars/ 100 MPa Operates at relatively lower pressure than UPLC Maximum backpressure between 300-400 bars 3 Inner diameter of the column is generally 0.75-1.8 mm Inner diameter of the column is 3-10 mm
- 4. Sr. No. UPLC HPLC 4 Dimension of column 150* 2.1 mm or 1mm (length* id) 2-15 cm length Dimension of column 150* 3.2 or 4.6 mm (length* id) 2-15 cm length 5 More selective and sensitive Less selective and sensitive 6 High resolving power Less resolving power
- 5. Sr. No. UPLC HPLC 7 Reduces process cycle time and assures end-product quality with reduced cost of operation and decreased run time. High cost of operation and high run time as compared to UPLC 8 It decreases the consumption of solvent and increases sample throughput More solvent consumption, less sample throughput 9 The higher back pressures compared to conventional HPLC decreases the life of the columns (column coagulation). Increasing the column temperature reduces the back pressure problem in UPLC. Less back pressure, more life of column
- 6. Sr. No. UPLC HPLC 10 The particles of less than 2 μm are mostly non-regenerable and, therefore, have a narrow use. Has more use 11 Higher precision in sample introduction is required Lower comparative to UPLC 12 Detectors uses small flow rates and low detection volume, Hence high data sampling rate is required. Detectors uses higher flow rates and high detector volume
- 7. Sr. No. UPLC HPLC 13 Stationary phase is modified to withstand high pressures. HYBRID of inorganic silica and organic group is used. Bridged ethylene hybrid (BEH), High strength silica (HSS), Charged surface hybride (CSH) are used as stationary phase. Inorganic Silica modified to different polarity is generally used. 14 Injection volume are small (generally 2 microliter) Injection volume are large (generally 5 microliter) 15 Less run time More run time
- 8. Sr. No. UPLC HPLC 16 More plate count is achieved during analysis Less plate count is achieved during analysis 17 System should withstand high pressures, more efficient pumps are required Less efficient pumps are required 18 Low extra column dispersion More extra column band broadening