Identification and Quantitative Determination of the IPH AGAG Peptide in Chewing Gum by Ultra-Efficient Liquid Chromatography

Ideal Pharma Peptide conducted chromatographic analysis of water extracts from chewing gum samples to identify and quantify the IPH AGAG peptide.

1 Research Methods

1.1 Study Objects

1.1.1 Description

A single product is a ball with a coating, weighing 1 gram and with a diameter of 11.5 mm.

1.1.2 Sample Preparation

The ball was rolled out on a foil substrate and the obtained mass was transferred quantitatively into a Vacuette tube. 5 ml of water was added, and the sample was extracted using a shaker for 15 minutes at a speed of 400 oscillations/min. Then the sample was centrifuged at 4000 rpm for 10 minutes, the extract was separated and introduced into the chromatograph. Three samples were prepared for analysis.

2 Chromatographic Analysis

The analysis was performed using ultra-efficient liquid chromatography (UPLC) with the following equipment:

• Acquity UPLC I-class chromatograph with a diode-array spectrophotometric (SP) detector (Waters, USA).

2.1 Chromatographic Conditions

Table 1

Quantitative assessments were made using the external standard method. An aqueous solution of the IPH AGAG peptide substance with a concentration of 10 µg/ml was used as the standard sample.

The content of the IPH AGAG peptide in a single piece of chewing gum weighing 1 gram (X) in µg was calculated using the formula):

Where S and S0 are the areas of the IPH AGAG peptide peaks on the chromatograms of the test and standard samples, respectively; C0 is the concentration of the standard sample solution in µg/ml, and 5 is the volume of the extract in ml.

3 Research Results

The quantitative analysis data are presented in the table.

Table 2. Content of IPH AGAG Peptide in Chewing Gum

Figure 1 – Chromatogram of the standard sample solution of AGAG peptide with a concentration of 10 µg/ml

Figure 2 – Chromatogram of chewing gum extract (Sample No. 2)