Which parameters (among linearity, LOD, LOQ, %RSD, % of recovery etc.) I should must check in HPLC method validation ?
Please look at the following below links which may help you in your analysis:
http://downloads.hindawi.com/journals/jamc/2018/1902510.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5345923/pdf/10.1177_1177390117690152.pdf
https://www.pharmaguideline.com/2017/12/hplc-method-validation.html
https://www.waters.com/webassets/cms/library/docs/720001826en.pdf
https://www.scielo.br/pdf/bjps/v45n1/10.pdf
Thanks
Yes. K prime, details of the instrument settings and many other parameters too. 'System Suitability' parameters should also be included as well. Please consult the appropriate regulations in your area for the minimum requirements, then exceed them as needed to show good chromatography fundamentals were followed. *Having a "Validated" method does not mean the method is in-fact valid or useful during an analysis. It only documents some of the steps and procedures used.
Hi! You can also check this example of HPLC method validation:
Article Development and validation of a liquid chromatographic metho...
It depends on the application of your method if you are doing assay method validation u does not require LOD/LOQ, while for the related substance you need to do all the above parameters.
Method validation is according to ICH Q2(R1) Methodology .
Dear MM Hasan,
Thanks for your interest. Actually for the validation of analytical method first of all you have to decide for which purpose you are intended to validate your method, for example- if you interested to develop and validate your method for the analysis of pesticides and veterinary drugs, I would suggest you to follow the EC documents (SANTE 12682/2019), and definitely you have to validate your method in terms of accuracy, precision, linearity, LOD and LOQ. Besides this, you have to check the matrix effects.
Dear M. D.H. Prodhan sir,
I want to detect and quantify Ascorbic Acid in a plant extract using an external standard. I have analyzed linearity, LOD, LOQ, % of RSD in terms of peak area and retention time, and % of recovery. Are these enough or not ?
Thanks for your query. I think the parameter that you have mentioned are adequate but you did not mentioned in which matrix you have selected for ascorbic acid determination. Another important thing is instrument, the best choice is to use LC-MSMS rather than HPLC.
Dear M. D.H. Prodhan sir,
I want to quantify by HPLC using a analytical reverse phase column. We don't have LC-MS in our laboratory.
Dear Hasan, don't worry about LCMS, let's start with HPLC. Wish you good luck
Thank you also, if you need any further assistance please communicate with me, I am happy to assist you.
Find these
1. Validation of microwave-assisted sample extraction-high performance liquid chromatography (HPLC) method for quantitative analysis of hydrolysed amino acids in aquatic biological samples
2. Validation of high-performance liquid chromatography (HPLC) method for quantitative analysis of histamine in fish and fishery products
Dear Mahmudul, listed parameters are sufficent for the establishment of the method (include specificity) but if you need to submit this method for regulatory agencies or adopt for the compendial monograph, then it require full validation as per ICH Q2R1 guidelines
Maybe these publication contain what you are looking for
doi:10.1016/j.jchromb.2008.12.056
or
Article Chromatographic methods development for clinical practice: r...
All mentioned plus Retention time stability, Gradient stability, from QC part ICV, CCV, LCS recovery etc.
- Badges
- Science method