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Identification, Analyitical Data Collection of Natural Products

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Identification, Analyitical Data Collection of Natural Products
Natural Products Experiment
Niki Robinson 10008562

Practical 2: Identification and quantification of taxifolin from milk thistle

Aim: to Identify and quantify the amount of taxifolin from three samples Silymarin stock solution, taxifolin and silybum marianum extract. `

Methodology: Using auto-pipette (20-100 microlitre), pipetted 0.06 mg of Taxifolin was added to 10ml conical flask and made up to the mark (10ml) with H2O.This procedure was repeated with the same amounts foe silymarin stock solution and silybum marianum extract. Taxifolin was transferred to 2ml brown bottle for HPLC auto sampler, this was also done to silymarin stock solution. Silybum marianum extract was filtered via a syringe filter (0.2 microlitre).The three samples were tested for concentrations of taxifolin via HPLC.This process was repeated a further 20 times with different amounts of taxifolin, silymarin stock solution and silybum marianum extract. Results of Taxifolin standard shown in table 1 and figure 1.

Results:
Table 1 Known Concentration of Taxifolin Standard Concentration mg/ml | Taxifolin peak area | 0.0036 | 1.876 | 0.0072 | 2.862 | 0.0144 | 4.518 | 0.0576 | 8.074 | 0.0576 | 15.273 |

Taxifolin Standard curve constructed from Table 1

Figure 1 Calibration curve Known Taxifolin Standard

Taxifolin Standard Curve Constructed from Table 1 with Error plus and minus bars

Figure 2 Taxifolin with Error Bars

The conditions of HPLC as the Taxifolin standard sample was run
HPLC Conditions, Halo C18, 4.6x150, 2um
Detection 280nM, injection volume 10mL
Table 2 HPLC Conditions Minute | Solvent A (Methanol) | Solvent B (0.5% Phosphoric Acid) | 0 | 20 | 80 | 2 | 20 | 80 | 20 | 60 | 40 | 25 | 60 | 40 | 30 | 20 | 80 |
The three unknown samples of Taxifolin standard (TS), Silymarin stock solution (SSS) and Silybum Marianum Methanol Extract (SBME), were diluted to 0.06 ml/10ml of distilled water. 10ml was then pipette small brown glass high

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