Table of Contents
What is calibration in HPLC?
The demonstration that a particular instrument or device produces results within specified limits by comparison with those produced by a reference or traceable standard over an appropriate range of measurements. Calibration of HPLC: Various Calibration. parameters are: ➢ Flow rate accuracy. ➢ Injector accuracy.
Which standard is used for calibration of HPLC?
Standard Operating Procedure (SOP)
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).
How do you calculate flow rate in HPLC calibration?
Collect the mobile phase at the detector outlet into a cleaned dried 25ml measuring cylinder (previously weight) for known time i.e. at 0.5ml/min for 10 min and calculate the flow rate /min by given formula. Flow rate per min = Volume dispense in selected time / Time.
What is calibrated range?
The calibration range is the interval comprising the measurement values possible when registered with a measuring device and typical for the respective measurement process. In time, within the calibration range there may be deviations for individual measurements.
What is SST in HPLC?
System suitability test (SST) is a test to determine the suitability and effectiveness of chromatographic system prior to use. The performance of any chromatographic system may continuously change during their regular use, which can affect the reliability of the analytical results.
What are HPLC parameters?
The HPLC parameters are the: efficiency factor(N), the retention factor (kappa prime), and the separation factor (alpha). Together the factors are variables in a resolution equation, which describes how well two components’ peaks separated or overlapped each other.
What is HPLC troubleshooting?
In an HPLC system, problems can arise from many sources. First define the problem, then isolate the source. Use Table 1 to determine which component(s) may be causing the trouble. A process of elimination will usually enable you to pinpoint the specific cause and correct the problem.
What is calibration of pH meter?
A pH calibration is the process of adjusting your pH meter by measuring solutions of a known pH value. This is because the characteristics of your electrode will change over time and this needs to be compensated for. A calibration does this by matching your pH meter to the current characteristics of your pH sensor.
What does HPLC stand for?
High-performance liquid chromatography
High-performance liquid chromatography (or high-pressure liquid chromatography, HPLC) is a chromatographic technique that can separate a mixture of compounds and is used in biochemistry and analytical chemistry to identify, quantify and purify the individual components of the mixture.
What is Principles of calibration?
Calibration Principles: Calibration is the activity of checking, by comparison with a standard, the accuracy of a measuring instrument of any type. It may also include adjustment of the instrument to bring it into alignment with the standard.
Why is uracil used in HPLC calibration?
Mixtures containing uracil are also commonly used to test reversed-phase HPLC columns. As uracil is essentially unretained by the non-polar stationary phase, this can be used to determine the dwell time (and subsequently dwell volume, given a known flow rate) of the system.
What are the parameters in HPLC calibration?
Flow rate accuracy test
What is the objective of calibration?
The purpose of calibration is to ensure that a measuring device is accurate, reliable and consistent. Any kind of measurement is used in a decision-making process, so it is essential that the data collected is accurate.
What is void volume in HPLC?
“Void Volume” refers to the HPLC Column void volume, “T zero time” or “dead time” (dead volume). This is a measure of the internal volume inside the column WITH the stationery phase particles. This volume is less than the calculated volume of a cylinder because the column contains porous particles.