High Performance Liquid Chromatography (HPLC)
Overview
High Performance Liquid Chromatography is commonly used to separate compounds in a mixture. The sample is moved through the column (stationary phase) by the mobile phase. The sample constituents elude the column at different times. These times typically dictate the analytes affinity to the column, hence determining the compounds in the mixture. The peak area from the spectra is then used to calculate quantitative amounts of the different compounds. HPLC is not only used to separate the compounds for analysis but can also be used to purify the mixture. But collecting the different elution’s at different times.
Services
- HPLC with ELSD (evaporated Light Scattering Detector) detection
- HPLC with RID (refractive index detector)
Pricing
- Expedited service: Starts from $1,000 per sample with 2 – 3 days turnaround.
- Regular service: Starts from $500 per sample with 7 – 10 days turnaround.
- For a comprehensive overview of our pricing structure, please log in to the Bee Portal.
Equipment
Agilent HPLC
- Features
- More chromatographic resolution from specially designed components in the sample flow path to achieve the lowest system dispersion.
- Higher peak capacity for challenging separations – switch easily between single dimension high-end UHPLC and the ultimate chromatographic power of 2D-LC
- Lower carryover for uncompromised data quality – multiwash capabilities of the Agilent 1290 Infinity II Multisampler reduce carryover to less than 10 ppm even for challenging compounds such as chlorohexidine.
- Unique detection capabilities – combine lowest detection limits with an ultrawide dynamic range by using the new Agilent 1290 Infinity II HDR-DAD or 1290 Infinity II ELSD
- Faster injection cycles with dual-needle injection – for higher sample throughput
- Higher sample capacity per bench space – up to 6144 samples within the footprint of a standard Agilent stack
- Flexibility for all applications – through wide power, temperature, an automatically scalable injection range, and gradient options
- Seamless transfer of methods between LCs, regardless of the brand – facilitated by intelligent system emulation technology (ISET) delivering unchanged retention time and peak resolution.
- Seamless integration in your chromatography data system – Agilent InfinityLab LC instrumentation runs smoother when controlled through third-party chromatography data systems such as Waters Empower or Thermo Scientific’s Chromeleon using the Agilent instrument control framework (ICF)
- Lower total cost of ownership – select from a variety of tools to achieve lowest cost-per-sample.
- Your choice for performance and sustainability: The 1290 Infinity II LC System has received the My Green Lab ACT (Accountability, Consistency, Transparency) label after an independent audit to verify its environmental impact throughout the product life cycle.
- Specification
- Column Capacity: 8
- Flow Range: up to 5 mL/min with G7120A and G7104A
- Injection Range: 0.1-100 µL (0.1 to 1500 µL with Multi Draw Kit)
- Maximum Number of Solvents: 4 (up to 26 with additional Solvent Selection Valves)
- Maximum Number of Temperature Zones: 2 with G7116B (1 with Vial Sampler with Integrated Column Compartment)
- Pump Type: Binary (Flexible)
- Special Features: Blend Assist with G7104A
- System Pressure Operating Range: up to 1300 bar
FAQ
A: HPLC is a tool that is used to determine what constituents are in the mixture. Based on the elution times and number of peaks in the spectrum will determine the different amounts compounds in the solution.
A: UPLC has the can separate the samples constituents at a much faster rate than HPLC by putting the system under vacuum.
A: HPLC works by passing a liquid sample through a column packed with a stationary phase material. A mobile phase, typically a solvent or solvent mixture, is pumped through the column at high pressure. Compounds within the sample interact differently with the stationary phase, leading to their separation into distinct peaks as they exit the column. A detector, such as UV-visible or fluorescence spectroscopy, measures these peaks, providing information about the compounds’ identity and concentration.
A: ELSD is useful in surfactant identification, which is molecules that are lacking functional groups.
A: Common detectors in HPLC analysis include UV-visible detectors, fluorescence detectors, refractive index detectors, and mass spectrometers. The choice of detector depends on the compounds being analyzed and the required sensitivity.
A: HPLC analysis is used in various applications, including pharmaceutical quality control, environmental monitoring, food and beverage analysis, clinical chemistry, and research in fields like biochemistry and materials science.
A: Retention time is the time it takes for a compound to elute from the HPLC column. It is a critical parameter used for compound identification. By comparing the retention time of an unknown compound to that of known standards, one can identify the substance.
A: HPLC achieves high performance and accuracy due to its ability to separate compounds in complex mixtures and its sensitivity in detecting and quantifying trace amounts of analytes. It also offers precise control over the mobile phase composition, flow rate, and column temperature.
A: Yes, HPLC is widely used for quantitative analysis. By measuring the area under the peaks in the chromatogram, HPLC can accurately determine the concentration of compounds in a sample when compared to calibration standards.
A: Challenges in HPLC analysis may include the need for appropriate sample preparation, selection of suitable columns and stationary phases, and the potential for interference from co-eluting compounds. HPLC may not be suitable for compounds that do not have adequate solubility in the chosen mobile phase.
A: HPLC analysis has revolutionized industries and research fields by providing a precise, efficient, and versatile tool for compound analysis. It has contributed to quality control in pharmaceuticals, environmental monitoring, and scientific advancements across numerous disciplines.