Total Curcuminoids Content by HPLC

Method for the estimation of  Piperine  content 

Reagent  Required :

Methanol  LR  Grade

Glass wares, Apparatus & Instruments  required  :

 Volumetric Flasks -          100mL, 50mL

 Beakers -                         100mL,50mL,

  U-V Spectrophotometer

Procedure:

Weigh  accurately about 200 mg of the Paprica oleoresin sample & transfer  into a 100 mL volumetric flask.Add approximately 50mL of methanol. Shake well until the Sample dissolves 

completely, warming if necessary. Make up the volume with methanol. Further dilute above 10mL solution to 50mL with methanol, 5mL solution to  50 mL with methanol, 5mL to 50mL with methanol in  volumetric flasks.Filter the solution if there is any suspended particle oservation.

Take the absorbance on U-V spectrophotometer at 342nm

Percentage of Piperine  content  is calculated as :

              Absorbance at 342nm

(1)     -----------------------------------    =     (A)

                 %  concentration

                   A x 285.33

 (2)   ------------------------                    =     (B)

                         10

                                                   B x 100

% content of piperine =    -----------------

                                                   32000

Method for the estimation of Total Guggulusterones  content 

Reagent  Required :

Methanol  LR  Grade

Glass wares, Apparatus & Instruments  required  :

 Volumetric Flasks         100mL, 50mL,250mL

 Beakers                       100mL,50mL,

 U-V Spectrophotometer

 Pippete –                     5mL,2mL

STANDARD PREPARATION:

Weigh accurately about 25 mg of working standard  into a 100mL volumetric flask. Add about 25mL of methanol. Ultrasonicate till completely dissolved and make up the  volume with methanol (Solution A). Further dilute 5mL solution A   to  50mL with methanol (Solution B). Further dilute 5mL solution B to 50mL with methanol.

SAMPLE PREPARATION:

Weigh accurately about 100 mg of sample into a 100mL volumetric flask. Add about 25mL of methanol. Ultrasonicate till completely dissolved and make up the  volume with methanol. warming if necessary.Make up the volume with methanol. Further dilute above 5mL solution to 50mL with methanol. So that the active  concentration in the sample is approximately similar to that of standard concentration.  Shake well and filter through what man No.1 filter paper.

Procedure :

Determine absorbance of both standard and sample solution at wavelength of 237.2nm.

Percentage of Total Guggulusterones content is calculated as:

                                              Absorbance of sample X Concentration of std  X   purity of std

  % of  Guggulusterones  =-------------------------------------------------------------------------------

                                                 Absorbance of standard    X  Concentration of sample 

For Raw Material :- Weigh accurately about 4.0gm Raw herb of Guggul in 250 mL R.B.Flask. add about 75mL of methanol and reflux for one hour. Cool and filter the solution. Rinse the residue in the flask with small quantity of methanol & filter through same filter paper and make up the volume with methanol. Further dilute above 5mL solution to 50mL with methanol.

Further dilute above 5mL solution to 100mL with methanol.

So that the active  concentration in the sample is approximately similar to that of standard concentration.  Shake well and filter through what man No.1 filter paper.

  

Method for the estimation of Curcuminoides   content 

Reagent  Required :

Acetone LR  Grade

Glass wares, Apparatus & Instruments  required  :

 Volumetric Flasks -       - 100mL, 50mL

 Beakers -                     - 100mL,50mL

 Spectrophotometer.

Procedure:

Take accurately about 60 mg of the Curcumin powder  in a 100 mL volumetric flask. Dissolve it completely in methanol and make up the volume up to the mark (Solution A). Take 5mL solution A into the 50 mL volumetric flask and make up the volume with methanol (Solution B). Take 2 mL solution B into the 50 mL volumetric flask and make up the volume with methanol. Shake well and take absorbance  on  425nm .Calculate  the %age of curcumin content taking E1% as 1680 & using methanol as blank.

Percentage of curcuminoides content may be calculated as :-

                           Abs x 100 x 50 x 50

% content   =  --------------------------------------

                          1680 x weight taken x 5 x 2

For Raw material :-  

Weigh accurately about 5 gm of sample  powder (20#) into a 250mL R.B. flask.Add 50 mL of methanol L.R. Grade and reflux on water bath for 30minutes.  Cool & filter into a    100mL volumetric flask. Repeat the process  in the same manner.Make up the volume to 100mL with the same solvent. Further dilute so as the absorbance should be around 0.420 to 0.450. Rest the procedure is same as above.