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Liquid Chromatography

AUTHORS
** conclusion or other key text **
Bin Yuan, Yanchao Shi,
Approx. 220Inc.
PerkinElmer Characters
Shanghai, China

Ferulic Acid Analysis

in Angelica Sinensis
Introduction
Radix by HPLC Angelica sinensis (A. sinensis) is the rhizome
of Angelica sinensis (Oliv.) Diels (Umbelliferae
family). It has been widely used not only as a health food and drug in Asia but
also as a dietary supplement in women’s care in Europe. Nearly 20% of the
traditional Chinese medicinal preparations included in the Chinese Pharmacopeia
contained A. sinensis. Many bioactive constituents including polysaccharides,
phenolic acids and angelica lactone have been isolated from A. sinensis.1
Among them, ferulic acid (Figure 1) is an important and valuable phenolic acid
whose content is used for assessing the quality of A. sinensis according to the
Pharmacopoeia of China.2

Figure 1: Chemical structure of ferulic acid (ChemDraw).

In this work, analysis of identified marker compound based on Chinese

Pharmacopoeia was analyzed using a PerkinElmer LC 300 with a PerkinElmer
Epic™ C18 column and consumables, and SimplicityChrom™ software (Figure 2).
A regulatory quality monitor workflow for analysis ferulic acid was established.
Application Brief: Ferulic Acid Analysis in Angelica Sinensis Radix by HPLC

Solvents, Standards and Samples

A ferulic acid reference standard material was purchased from
National institutes for food and drug control (NIFDC), China.
The herbal plant sample was purchased commercially from
a pharmacy store. All solvents were LC/MS grade. All other
chemicals and reagents were of the highest grade available.

Preparation of sample solution: transfer 0.2 g sample powder

to a 100 mL flat bottom conical flask, add 20 mL of 70% (v/v)
methanol aqueous solution. Reflux under boiling-water bath for 30
min. The solution was filtered with 0.22 µm PTFE syringe filters
(P/N: 02542884).

Preparation of standard solution: dissolve a known quantity of

ferulic acid in 70% (v/v) methanol aqueous solution to a final
concentration of 10 μg/mL.

Figure 2: The schematic diagram of the LC total workflow solution.

Experimental
Hardware/Software
The chromatographic separation was conducted by a
PerkinElmer LC 300 UHPLC system and detection was
achieved using a PerkinElmer LC 300 PDA detector. All
instrument control, data acquisition and data processing were
performed using the SimplicityChrom software.
Method Parameters
Results and Discussion
Based on the chromatographic conditions and system suitability
specification in the Chinese Pharmacopoeia, the standard solution
and sample solution were analyzed using a PerkinElmer LC 300
with an Epic C18 column.
Figure 3 shows that Epic C18 has excellent separation capability
on the identified marker compound with good retention and
peak shape. Excellent efficiency was observed for ferulic acid
(14360 plates) with the plate number specification (not less than
5000 plates) being met.

The LC parameters are shown in Table 1.

Table 1: LC method parameters.

Epic C18 150×4.6 mm, 3 µm

Column
(P/N: 135191-EC18)
A: 0.085% H3PO4
Mobile Phase B: Acetonitrile
A/B: 83:17

Flow Rate 1.0 mL/min

o
Oven Temperature 35 C

Analytical: 316 nm (Bandwidth: 4 nm)

Detector Wavelength
Reference: 500 nm (Bandwidth: 4 nm)
Figure 3: The chromatograms of sample (top) and standard (bottom).
Sampling Rate 5 pts/sec

Injection Volume 10 μL (partial loop) Six consecutive standards were analyzed and the retention
and area response generated excellent repeatability, as
shown in Table 2, with relative standard deviations (RSD %)
below 1.0%.

www.perkinelmer.com 2
Application Brief: Ferulic Acid Analysis in Angelica Sinensis Radix by HPLC

Table 2: Repeatability results of six injections of standard (10 μg/mL).

Time (min) Peak Area (mAU*s)

RSD% 0.10 0.38

Conclusion
• A robust and repeatable LC-UV workflow for the analysis
of ferulic acid in Angelica sinensis has been developed
by coupling a PerkinElmer LC 300 system with an Epic
C18 column.
• This method can be applied for determineration of ferulic
acid in accordance with the Chinese Pharmacopoeia.
• The Epic C18 column shows excellent separation capability
and peak shape due to the superior base deactivation.

References
1. Hongwei Wu et al. Molecules 2020, 25(15), 3356
2. Vol. 1, Chinese Pharmacopoeia 2020.

Consumables
Component Description Part Number

Columns Epic C18, 150 x 4.6 mm, 3 µm 135191-EC18

2 mL Amber 9 mm Screw Top Vial with Write-on
HPLC Vials N9307802
Patch and Fill Lines (100/Pack)
9 mm Screw Top Blue (Polypropylene) Cap with
HPLC Vials Caps N9306203
PTFE/Silicone Pre-Slit Septa (100/Pack)
Peek Fittings Finger-Tight Fittings, PEEK, 5.5K psi Max (5/Pack) N9307822

Stainless Steel Fittings Ti Hybrid w/Flat Wrench Ferrule/Nut N9306301

Syringes Syringe 1 mL BD Luer-Lok Disposable (100/Pack) 02542890

Syringe Filters 0.22 μm PTFE (Hydrophobic) Syringe Filter, 17 mm 02542884

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