Food nutritional fortifier raffinose detection

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A. 1 General regulations

Unless otherwise specified in this standard, the purity of the reagents used should be above analytical purity. The standard titration solution used, impurity determination standard solution, preparation, and products should be prepared in accordance with the provisions of GB/T601, GB/T602, and GB/T603. The test water should comply with the provisions of GB/T6682. The solution used in the experiment refers to aqueous solution when no specific solvent is specified for preparation

A.2 Determination of raffinose content

A.2.1 Reagents and materials

A.2.1.1 raffinose reference substance: purity ≥ 99%

A.2.1.2; Acetonitrile: Chromatically pure

A.2.2 Instruments and Equipment

A.2.2.1 High performance liquid chromatography: equipped with a differential refractive detector or other equivalent detector.

A.2.2.2 Ultrasonic oscillator

A.2.3 Reference chromatographic conditions

A.2.3.1 Chromatographic column: Gum bonded column (amino bonded phase column)

A.2.3.2 Mobile phase: acetonitrile+water=75+25, this ratio moderately increases or decreases depending on the separation condition

A.2.3.3 Column temperature: 40 ℃

A.2.3.4 Flow rate: 0.8mL/min, which increases or decreases moderately based on the inner diameter of the column and the separation condition

A.2.3.5 Injection volume: 10uL

A.2.4 Analysis Steps

A.2.4.1 Preparation of Standard Solution

Weigh approximately 0.05g of raffinose reference substance, accurate to 0.0001g, place it in a 10mL volumetric flask, dissolve it in water, dilute to the mark, and obtain the raffinose standard solution.

A.2.4.2 Preparation of Sample Solution

Weigh approximately 0.5g of the sample to the nearest 0.0001g, place it in a 50mL volumetric flask, dissolve it in water, and dilute to the mark.

. The solution was subjected to ultrasonic treatment for 10 minutes, filtered with a 0.45um filter membrane, and the filtrate was used for backup

A.2.4.3 Determination

Under the reference chromatographic conditions in A.2.3, perform chromatographic analysis on the standard solution and the sample solution, respectively.

. Record the peak area values of raffinose in the chromatograms of the standard solution and the sample solution

A.2.5 Result Calculation

The mass fraction w1 of raffinose content (calculated on a dry basis) is calculated according to equation (A.1):

Where:

cs - the concentration of raffinose in the standard solution, in grams per milliliter (g/mL)

V1- Volume of the sample solution, in milliliters (mL)

m1- The mass of the sample in the sample solution, in grams (g)

w0- Drying loss of the sample,%

A1- peak area value of raffinose in the chromatogram of the sample solution

As - peak area value of raffinose in the standard solution chromatogram

 

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