Determination of Tyrosol in Wine by High Performance Liquid Chromatography

Determination of Tyrosol in Wine by High Performance Liquid Chromatography

1. Instrument Configuration and Experiment Methods

1.1 Instrument Configuration

1.2 Experiment Method

1.2.1 Reagent Preparation

1.2.1.1 Tyrosol standard stock solution (1000mg/L): Take the appropriate amount of tyrosol standard, dissolve and Fix the volume with methanol, a standard stock solution with a concentration of 1000mg/L will be prepared, sealed and stocked at -4℃.

1.2.1.2 Tyrosol standard working solution: Pipette the appropriate amount of tyrosol standard stock solution accurately, dilute with methanol to form a series of working curves with concentrations of 0.1mg/L, 1mg/L, 1.5mg/L, 3mg/L, 5mg/L, 7.5mg/L, 10mg/L respectively.

1.2.2 Chromatography Conditions

Table 3 Chromatography Conditions

Table 4 Proportion of Mobile Phase

1.2.3 Sample Pretreatment

Take appropriate amount of white wine samples, through the 0.45μm microporous filter membrane, then to be measured.

2. Experimental Result

2.1 System Suitability

Fig 1 Chromatogram of 10mg/L Standard

Table 5 10mg/L Standard Test Data

Note: From the chromatogram and data, it can be seen that the tyrosol peak shape is good, there are no other peaks around the target peak, and the theoretical plate number is high, which meets the experimental requirements.

2.2 Standard Curve

                                       Fig 2 Test Result of Standard Curve

Note: From the above chromatogram, it can be seen that the correlation coefficient value R of tyrosol curve is above 0.999, which meets the experimental requirements.

2.3 Repeatability

Fig 3 Repeatability Chromatogram of 3.75mg/L Standard for 6 Injections

Table 6 Repeatability Test Data of 6 injections for 7.5mg/L Standard

Note: According to the above table data, it can be seen that the RSD of tyrosol retention time repeatability is 0.053% and the RSD of peak area repeatability is 0.485%, both of which has good repeatability. It meets the experimental requirements.

2.4 Detection Limit

Fig 4 Test Chromatogram of 0.1mg/L Standard

Table 7 Test Data of 0.1mg/L Standard

Note: According to the above table data, the detection limit of tyrosol is 0.0073mg/L with 3 times signal-to-noise ratio, which meets the experimental requirements.

2.5 Test Results of a Brand of White Wine

Fig 5 Test Chromatogram of a Brand White Wine

Table 8 Test Data of a Brand White Wine

Note: 0.275 mg/L of tyrosol was detected in a brand of white wine.

2.6 Spiked Testing Result of a Brand White Wine

Fig 6 Spiked Testing Chromatogram of a Brand White Wine

Table 9 Spiked Test Data of a Brand White Wine

Note: Add 15μL of 100mg/L standard in a 1mL white wine, and according to the detection concentration of white wine and the spiked concentration, the theoretical concentration is 1.775 mg/L. From the detection concentration in the above table, the spiked recovery is 101.4%, which meets the experimental requirements.

2.7 Attentions

The Tyrosol Standard stock solution needs to be stored at low temperature, otherwise its content will decrease.

3. Conclusion

This article introduces the determination of tyrosol content in white wine by Wayeal high performance liquid chromatograph LC3210 series equipped with ultraviolet detector. The experimental results showed that the peak shape of tyrosol is good in the system adaptability test, and there are no other peaks around the target peak, and the theoretical plate number is high, which met the experimental requirements. The curve correlation coefficient R value is above 0.999. The RSD of tyrosol retention time is 0.053%, and the RSD of peak area is 0.485%, which is good reproducibility. The detection limit of tyrosol is 0.0073mg/L. The recoveries are 101.4% with the spiked 1.5mg/L in the sample. The results of the above data meet the requirements of the instrument for the test method.