Can you recommend a method for preparation in order to quantitatively determine vitamin B6, 3,12 in pharmaceutical multivitamin ampules by HPLC?
Dissolve the sample in a polar solvent which you will use has your mobile phase, run your Vitamin standards using the same solvent. If you HPLC has good detector you should get the chromatograms which you can calculate the quantities
Here is a method jut published in the J AOAC Int
J AOAC Int. 2012 Jan-Feb;95(1):74-83.
Development and validation of rapid ion-pair RPLC method for simultaneous determination of certain B-complex vitamins along with vitamin C.
Patil SS, Srivastava AK.
SourceUniversity of Mumbai, Department of Chemistry, Vidyanagari, Santacruz (East), Mumbai 400098, India.
Abstract
A rapid, simple, and accurate ion-pair RPLC method has been developed for simultaneous analysis of vitamin C and major B-complex vitamins. An RP C18 column thermostated at 30 degrees C was used with gradient elution of mobile phase comprising 10 mM potassium dihydrogen phosphate buffer (containing 3 mM sodium hexane-1-sulfonate, adjusted to pH 2.80 with o-phosphoric acid) and methanol at a flow rate of 1.0 mL/min to achieve the best possible separation and resolution of all vitamins in about 11.00 min. The detection was performed at 274 nm. The method has been implemented successfully for simultaneous determination of vitamins present in 12 multivitamin/multimineral pharmaceutical preparations, as well as in human urine. Typical validation characteristics were evaluated in accordance with International Conference on Harmonization guidelines. Good linearity over the investigated concentration levels was observed. Intraday repeatability was < or = 2.0%, and interday variation was < or = 2.6%, for all vitamins. The method can be used for assay of these vitamins over a wide concentration range with good precision and accuracy; hence, it would be appropriate for routine QC as well as in clinical analysis.
0.1N HCL with different ratio of Methanol or it can proceed with 0.1N Methanolic HCL to extract multivitamins(Niacin, Niacinamide, Vitamin B1, B2,B6, Vitamin C and Folic ) from ampoules.
Vitamin C and Folic acid can also be extract by ratio of methanol and 0.1% Glacial acetic acid (50:50)%v/v.
For quantifying all the vitamins HPLC/UPLC could be the useful tools.
@ chinmoy thanks alot , what about HPLC condition at this case , do you have reference
Sorry mario , i can't understand you , could you please write in english?
Salma Remember that vitamin C was oxidized in air. Stabilize the sample with vitamin E. excuse me bad english
Unfortunately, i can't to help you, because it isn't the area of my specialisation. I am a family physician , and i do only a clinical researches.
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