Is anyone expert in RP-HPLC method development and quantification
What do you mean with "ideal and simple equation"? I think we can do assay of most of pharmaceuticals by using RP-HPLC; you can search the method of HPLC determination for any pharmaceuticals in Google, Scopus, or other data base. If you have the column, mobile phase and suitable detector (UV, DAD,ELSD, MS or others), than you can apply in your laboratory, after optimization you can do validation of the method.
Hi Sarif,
If I am not wrong, then I guess you are searching this equation to find out the assay result of your sample preparation:
Assay = (Peak area of sample / Peak area of standard) X (Amount of standard taken / volume of standard solution) X (Volume of sample solution / Amount of sample taken) X (Potency of standard / 100) X Dilution factor X Conversion factor
AT1 x Ws x 20 x P
% Assay = -------------------------------------
AS x 20 x V1 x L.C
Where,
AT1= Peak area counts of the test preparation.
AS = Average peak area counts the standard preparation.
P= Percentage Potency of standard
Ws = Weight of working standard taken in mg.
WTS= Weight of net content taken for test preparation.
V1 = Volume of test solution in mL.
L.C = Label claim in mg per mL.
Dear Sarif,
Do you mean equation above as "equation for calculation of sample"?
For using equations that described by Subrata and Harshal above (=single point calibration) to calculate the concentration of sample/test, some requirements should be fulfilled (see the book from J. Ermer & P. Nethercote (Eds.) Method Validation in Pharmaceutical Analysis, Wiley-VCH, 2015, pp.146, 158). See Table 5.14
1. Linear regression function should be proved (r, Vxo, Mandel etc should have acceptance value)
2. Neglige constant systemic errors (ordinate intercept)
3. Homogeneity of variance (homoscedasticity)
If 1, 2, 3 were not fulfilled so single point cannot be applied.
You can also determine the concentrations of sample/test using calibration curve (by external or internal calibration).
I recommend you to read the book of Ermer & Nethercote (I think you can get via ResearchGAte).
You can read the method development in different kinds of HPLC techniques, especially haw you can manipulate the practical factors basing upon some theoretical factors such as retention factor k, selectivity factor, and number of theoretical plates N, you can find these topics in detail in Quantitative chemical analysis by D. C. Harries.
% Assay= (area of sample/area of std)*(conc of STD/Conc of Sample)*(Lable claim/Avg. wt of formulation)*(potency/100)*100
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