Fructus forsythia is a Traditional Chinese Medicine with property of bitter, cool, and non-toxic. The major functions are clearing away heat and toxic materials, eliminating stagnation and reducing swelling. The product is used to alleviate the symptoms caused by Diseases epidemic febrile disease, erysipelas, macula, carbuncle, scrofula and dysuria.
Table 1 Information of Analytes
Material and Reagents
Watsons water; HPLC grade of acetonitrile;
Fructus forsythia: provided by customer.
Transfer 1.0 g powdered fructus forsythiae and 15 mL methanol into a conical flask with cover, check the weight and impregnate overnight. Sonicate for 25 min ultrasonic treatment and keep it to the ambient. Weigh it again and add methanol for the loss weight. Evaporate 5.0 mL extract to dryness, add 0.5 g neutral alumina on neutral alumina column (from 100~120 mesh, 1g, inner diameter of 1~1.5 cm), eluted with 80mL 70% absolute ethyl alcohol. Dry the eluent under nitrogen flow and dissolve the residue with 50% methanol-water. Transfer it into 5.0 ml volumetric flask, dilute to scale, shake well. Filter the extract through 0.45 mm filter followed by HPLC analysis.
Transfer 2.0 mg reference substance of forsythin into a 10 mL volumetric flask, add methanol into it, dissolve and dilute to scale as the reference solution.
HPLC Column: Venusil® XBP C18(L), 5 μm, 150 Å, 4.6 × 250 mm;
Mobile phase: water : acetonitrile = 75:25 (V/V);
Detection: 277 nm;
Flow rate: 1.0 mL/min;
Column temperature: 30 °C;
Injection: 10 mL.
Results and Discussion
Venusil® XBP C18(L) column was adopted to detect the Forsythin in Fructus forsythia. The result described that excellent separation was obtained in complex TCM matrix with sharp and symmetric peak shape.
Table 1 Performance of Venusil® XBP C18(L) in analysis of Forsythin in Fructus forsythia
Forsythin Reference Solution
Extract of Fructus forsythia
Figure 1 Chromatogram of Forsythin Reference Solution using Venusil XBP C18(L)
Figure 2 Chromatogram of the extract using Venusil XBP C18(L)
A robust HPLC method to analysis the Forsythin in Fructus forsythia was introduced in this paper. Venusil XBP C18(L) was adopted and excellent separation was gained with high resolution. The result shows that the Venusil XBP C18(L) was suitable for the determination of Forsythin without fear of the complex TCM matrix influence.
Venusil® XBP C18(L)
5 μm,150 Å, 4.6 × 250 mm
5 - 70 °C; Max. 2 columns of 300mm
1.5 mL vials
Screw neck vials, 12 × 32 mm
Caps and Septa
Screw neck cap, center hole; red silicone/ white PTEE septa, slitted
Syringe Filter (Nylon)
Monofilm, 13 mm, 0.22 μm
Disposable Needle-Free injection systems
2 mL, 100/pk