High Performance Liquid Chromatography (HPLC) is a powerful analytical technique for the separation, identification, and quantitation of a wide variety of chemicals, HPLC is widely used in a wide variety of fields, including pharmaceutical, food, and environmental.
Basic Principles of HPLC
HPLC uses a liquid mobile phase (solvent) and a stationary phase (packing material in the column).The sample is dissolved in the mobile phase and injected into the column under high pressure. Through interaction with the stationary phase in the column, the components of the sample pass through the column at different rates and are separated by the components. Each of these separated components is detected by a detector at the exit of the column and data is recorded.
Main components of HPLC
- Pump: A pump used to pump mobile phase through a column at a constant flow rate.
- Sampler: A device that injects an exact amount of sample into a column.
- Column: A tube filled with stationary phase. This is where the components are separated.
- Detector: A device that detects the components coming out of a column, typically a UV detector or mass spectrometer (MS).
- Data system: A computer system that analyzes the detected signals and outputs them as chromatograms.
Types of HPLC
HPLCの方法は主に以下のように分類されます：
- Reverse Phase Chromatography: The most common form of HPLC, used to analyze non-polar to moderately polar components.
- Normal Phase Chromatography: A highly polar stationary phase with a polar solvent as the mobile phase used to analyze polar molecules.
- Ion-exchange chromatography: a method used to separate molecules with a charge, where the column is made of a material capable of exchanging cations or anions.
HPLC Applications
- Drug analysis: verification of drug purity and content, identification of metabolites, etc.
- Food Analysis: Analysis of additives, pesticide residues, and nutritional components in food products.
- Environmental analysis: detection of contaminants in water and soil.
HPLC has become an indispensable technology in many scientific and industrial fields due to its high sensitivity and accuracy.
Cost and Time
The cost and time required to analyze a drug’s components using high-performance liquid chromatography (HPLC) depends on a variety of factors. Below is a summary of the major factors and estimates.
Complexity of Analysis
- Type of component to be analyzed: depends on whether it is a single component or multiple components. Simultaneous analysis of multiple components is technically complex and increases time and cost.
- Sample complexity: Pretreatment effort varies depending on whether the sample is pure, contains multiple components, or has a complex matrix, such as a biological sample.
Necessity of pretreatment
- Sample Preparation: Sample preparation (extraction, concentration, filtration, etc.) required prior to HPLC analysis may require additional time and cost.
Analysis Time
- Analysis Run Time: The actual run time for HPLC is typically several minutes to several tens of minutes. However, column and mobile phase conditions may vary depending on the analytes to be analyzed and the purpose of the analysis, and the time may increase accordingly.
- Total analysis time: It typically takes several hours to a day per sample, including setup, sample preparation, the actual HPLC run, and data analysis.
Cost
- Analytical Cost: The cost of an analysis using HPLC varies depending on the laboratory and country in which it is performed, but generally ranges from a few thousand yen to several tens of thousands of yen. Costs may vary for analysis of large numbers of samples for research purposes or for routine analysis in commercial trials.
Other Factors
- Laboratory capacity and expertise: The equipment of the laboratory where the analysis is performed and the expertise of the technicians also affect cost and time.
- Urgency: If results are urgent, additional fees may apply.
To obtain a specific estimate, it is important to contact directly the laboratory you plan to request the analysis from and clarify the sample details and analysis requirements. Based on this, they can provide you with an accurate estimate and time projection.
