Table of Contents
What are the calibration parameters of HPLC?
HPLC Calibration Procedure
| Parameters | Test Condition |
|---|---|
| HPLC Column | ODS C18, (25 cm x 4.6 mm ID, 5 µm) |
| Flow Rate | 1.0 ml / min. |
| Wavelength | 254 nm |
| Range | 1.0 AUFS |
Which standard is used for HPLC calibration?
Standard Operating Procedure
This HPLC calibration Standard Operating Procedure (SOP) describes all the individual steps necessary for calibrating a HPLC in accordance with cGMP (current good manufacturing practices).
What does calibration refer to in HPLC?
Calibration refers to the process of determining the relation between the output (or response or signal) of a measuring instrument and the value of the input quantity or property.
Why is caffeine used in HPLC calibration?
Caffeine is convenient because it is not volatile, readily available, safe, has strong UV adsorption if you are using UV detector, and does not retain too long on RP columns (faster calibration).
What is wavelength accuracy in HPLC?
Depending on the regulations or. guidelines applied, most authorities require accuracy to be within 2 to 3 nm of a certified. standard within the range used. In practice, we generally achieve accuracy of equal to or. better than 0.5 nm across a range of UV / VIS wavelengths.
What are HPLC standards?
HPLC standards are defined reference compounds or materials that test the quality, purity, and identity of a particular test sample by comparison. Ready-to-use HPLC standards can reduce cost, streamline production processes and ensure reproducibility in research and development.
What is SN ratio in HPLC?
Signal-to-noise often is used to help determine the limit of detection or limit of quantification of an HPLC method. Most workers calculate S/N just as the ratio of the signal to the noise, so S/N = 367/66 = 5.56 in this example.
Why is anthracene used in HPLC calibration?
Anthracene has a secondary maximum at 350 nm. A programmable variable wavelength detector is capable of scanning over a narrow range, just not quickly enough to be useful for real-time spectra collection.
How do I calibrate my HPLC with caffeine?
Weigh and transfer about 50.0 mg of caffeine working standard to a 100 ml volumetric flask. Add 60 ml Mobile Phase, dissolve with the help of sonication and make up the volume with the same solvent (500 ppm). Dilute 2 ml of the resulting solution to 50 ml with Mobile Phase (20 ppm).