Identification of Citri Reticulatae Pericarpium (Chenpi) From Different Cultivars via LC–MS/MS and UPLC Coupled With Multivariate Chemometrics Analysis

TL;DR

This study uses advanced chemical analysis and statistical methods to distinguish different types of Citri Reticulatae Pericarpium based on their unique flavonoid profiles.

Contribution

The study introduces a rapid and reliable method combining UPLC and chemometrics for cultivar differentiation and quality control of Citri Reticulatae Pericarpium.

Findings

Nine flavonoids were identified as key markers for differentiating seven CP cultivars.

Hesperetin was found exclusively in the 'Chachi' cultivar, indicating its potential as a specific marker.

Multivariate analysis successfully grouped and separated the cultivars based on their chemical profiles.

Abstract

Citri Reticulatae Pericarpium (CP), derived from the peel of
Citrus reticulata
Blanco and its cultivated varieties, is widely used and has substantial commercial value. However, notable differences in origin, quality, and price are observed among various CP cultivars. To ensure effective quality control and safe clinical use, a reliable and efficient method for distinguishing these cultivars is urgently needed. This study aimed to investigate the chemical profile differences among seven CP cultivars and to establish a rapid and accurate multi‐component quantitative method using ultra‐high‐performance liquid chromatography (UPLC) for their differentiation. Multivariate statistical analyses, including PCA, OPLS‐DA, and HCA, were employed to distinguish the seven cultivars. LC–MS/MS analysis identified 53 compounds, including 48 flavonoids, and nine representative flavonoids were selected for quantitative determination. The developed UPLC‐based method exhibited high specificity, linearity, precision, repeatability, stability, and accuracy. PCA revealed that
C. reticulata
“Chachi” (GCP),
C. reticulata
“Succosa” (BDZ),
C. reticulata
“Tankan” (JG), and
C. reticulata
“Unshiu” (WZMG) formed distinct clusters, indicating substantial differences in flavonoid composition. In contrast,
C. reticulata
“Kinokuni” (NFMJ),
C. reticulata
“Ponkan” (PG), and
C. reticulata
“Dahongpao” (DHP) grouped together but were further differentiated by PCA and OPLS‐DA, consistent with the HCA results. Hesperetin was quantified exclusively in GCP, suggesting its potential as a cultivar‐specific marker. Although the overall chemical profiles of the seven CP cultivars were similar, their flavonoid contents varied significantly. The combined UPLC quantification and multivariate analyses provide an effective approach for distinguishing major CP cultivars and offer new insights for quality control and future research.

A UPLC‐based quantitative analysis combined with multivariate chemometric methods differentiated seven Citri Reticulatae Pericarpium cultivars based on significant variations in nine flavonoids. Chemometric analysis effectively separated the seven cultivars, and hesperetin was identified as a unique marker for
C. reticulata
“Chachi”. This integrated analytical approach provides a rapid and reliable strategy for cultivar identification and quality control.

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