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. 2019 Sep 23;24(19):3455.
doi: 10.3390/molecules24193455.

A Simple, Rapid, and Practical Method for Distinguishing Lonicerae Japonicae Flos from Lonicerae Flos

Fang Zhang  1 , Pengliang Shi  2 , Hongyan Liu  3 , Yongqing Zhang  4 , Xiao Yu  5 , Jing Li  6 , Gaobin Pu  7
Affiliations

A Simple, Rapid, and Practical Method for Distinguishing Lonicerae Japonicae Flos from Lonicerae Flos

Fang Zhang et al. Molecules. .

Abstract

Lonicerae japonicae flos (LJF), the dried flower buds of Lonicera japonica Thunb., are often adulterated with Lonicerae. flos (LF), which is derived from the other four Lonicera species. Scholars at home and abroad have established several analytical methods to distinguish LJF from the four Lonicera species of LF; however, to date, no effective and practical method has been established for distinguishing LF from LJF. In our present study, the HPLC fingerprints of LJF and LF were compared, and differences in the content of one of the iridoids were found. Column chromatography combined with pre-HPLC was used for isolating and preparing the iridoid, and its structure was identified as secologanic acid. Then, a method for determining the content of secologanic acid was established using HPLC. The amounts of secologanic acid in 34 batches of LJF and 38 batches of LF were determined. The average amount of secologanic acid in 34 batches of LJF was 18.24 mg/g, with values ranging from 12.9 mg/g to 23.3 mg/g, whereas the average amount in 38 batches of LF was 1.76 mg/g, with values ranging from 0.2 mg/g to 7.2 mg/g. Therefore, secologanic acid can be considered as one of the characteristic components for distinguishing LJF and LF. Our study not only provides a rapid, simple, sensitive, and practical method for identifying LJF and LF but also establishes a method for discovering the characteristic components of other herb-medicines that are susceptible to adulteration.

Keywords: Lonicerae flos; Lonicerae japonicae flos; high performance liquid chromatography; multiple statistical analysis; secologanic acid.

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Conflict of interest statement

The authors report no conflicts of interest.

Figures

Figure 1
Figure 1
Chemical structures of the representative components in flower buds of Lonicerae species (1–14).
Figure 2
Figure 2
The overlapping fingerprints of L. japonicae flos (LJF) and LF (a) and the UV spectrum of compound S in LJF (b). Column: An Agilent ZORBAX XDB C18 (4.6 × 250 mm, 5 μm) column; column temperature, 30 °C; mobile phases: phosphoric acid solution (A) and acetonitrile (B) (5–15% B at 0–20 min, 15–55% B at 20–45 min, and 55–100% B at 45–60 min); flow rate: 1.0 mL/min; detection wavelength: 254 nm; injection volume: 10 µL.
Figure 3
Figure 3
HPLC chromatograph of eluents eluted with H2O and ethanol of different concentrations on a macroporous resin. The chromatographic conditions were the same as those described in the HPLC fingerprint.
Figure 4
Figure 4
The Q Exactive Hybrid Quadrupole-Orbitrap-MS spectrum of compound S in positive mode.
Figure 5
Figure 5
The content of secologanic acid in LJF under different extraction conditions: (a) solvent volume, (b) extraction time, (c) and extraction method.
Figure 6
Figure 6
The HPLC chromatograms of standard (a) and samples. (b) LJF sample; (c) LF sample; S: secologanic acid. Column: An Agilent ZORBAX XDB C18 (4.6 × 250 mm, 5 μm) column; column temperature, 30 °C; mobile phases: 0.1% phosphoric acid solution (A) and acetonitrile (B), using a gradient elution of 5–15% B at 0–20 min, 15–23% B at 20–25 min, 23–95% B at 25–26 min, and 95% B at 26–30 min; flow rate: 1.0 mL/min; detection wavelength: 240 nm; injection volume: 10 µL.
Figure 7
Figure 7
Scatter plot acquired from LJF and LF samples (a) and the contents of secologanic acid in LJF and LF (b) (x-axes show LJF and LF, the different sample; y-axes have the content of secologanic acid).
See this image and copyright information in PMC

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