Determination of Salidroside in Pharmaceuticals by High Performance Liquid Chromatography（HPLC）

Abstract

Purpose: Determination of Salidroside in Pharmaceuticals by High Performance Liquid Chromatography（HPLC）

Method: C18 column, 4.6*250mm, 5μm;

Wavelength: 275nm;

Mobile Phase A: Water; mobile phase B: methanol;

Flow Rate 1.0ml/min;

Temperature: 30°C;

Injection Volume: 5μl.

A standard curve was established and the content of the target was calculated by the external standard method.

Keywords: HPLC, UV Detector, Herbal, Salidroside

1. Experiment Method

1.1 Instrument Configuration

Wayeal LC3200 Series HPLC

Table 1 System Configuration of HPLC

1.2 Test Conditions

Column: C18, 5μm, 4.6*250mm

Temperature: 30°C

Wavelength: 275nm

Flow rate: 1.0ml/min

Mobile phase: A: water; B: methanol

Injection volume: 5μL

Gradient Condition:

1.3 Instrument, Reagents and Consumables

Reagents: ultrapure water, Methanol(GR)

Standards: Salidroside (99.7%)

Auxiliary device: chemical balance; solvent filter; ultrasonic cleaners

Experimental materials: filter membrane: aqueous phase filter membrane 0.45μm

1.4 Preparation of Solutions

1.4.1 Standard Solutions: Take appropriate amount of salidroside standard into a volumetric flask and dissolved in methanol to make the concentration of 0.0084125mg/mL, 0.016825mg/mL, 0.03365mg/mL, 0.0673mg/mL, 0.1346mg/mL, 0.2692mg/mL, 0.673mg/mL.

1.4.2 Sample Preparation: Take 1.0022g of sample 1 into a volumetric flask, add methanol and dissolve to 25mL. Take 1.0794g of sample 2 into a volumetric flask, add methanol and dissolve to 25mL.

2 Results and Discussion

2.1 System Suitability

Fig 1 Chromatogram of Salidroside Standard

Table 2 Chromatography Parameters of Salidroside Standards

Analysis: The test results of salidroside were good with symmetrical peaks and high theoretical plate number.

2.2 Standard Curve

Fig 2 Superimposed chromatogram of Salidroside Standard solutions

Fig 3 Curve Equation and Correlation Coefficient of Salidroside Standard Solutions

Analysis: The linear range of salidroside standard curve is good, r>0.999.

2.3 Repeatability

Fig 4 Repeatability Chromatogram of Salidroside Standards (n=6)

Table 3 Repeatability Chromatographic Parameters Table of Salidroside (n=6)

Analysis: 6 injections of 0.2692 mg/L salidroside show good reproducibility, and the RSD value of retention time is 0.055% and the RSD value of peak area is 0.340%.

2.4 Sample 1

Fig 5 Chromatogram of Sample 1

Table 4 Chromatography Parameters of Sample 1

Analysis: The content of salidroside in the sample 1 was 0.061933 mg/L, which was calculated according to the standard curve equation.

2.5 Sample 2

Fig 6 Chromatogram of Sample 2