Crops ›› 2025, Vol. 41 ›› Issue (1): 202-207.doi: 10.16035/j.issn.1001-7283.2025.01.025

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Simultaneous Determination of Contents of Five Kinds of Soluble Sugars in Codonopsis pilosula during Dormancy by HPLC-ELSD Method

Tang Shunli(), Zhang Yanhong(), Du Tao, Chen Hui, He Chunyu, Dong Wanqi   

  1. Department of Pharmacy, Gansu University of Chinese Medicine, Lanzhou 730000, Gansu, China
  • Received:2023-05-20 Revised:2023-07-25 Online:2025-02-15 Published:2025-02-12

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Abstract:

A method was developed for the simultaneous determination of contents of five soluble sugars in different organs of Codonopsis pilosula (Franch.) Nannf. during natural dormancy by HPLC-ELSD for investigating the role of soluble sugars in the cold resistance. The suitable detection conditions for the five soluble sugars were: Analysis Column (4.6 mm×250 mm 5-Micron), acetonitrile:water (70:30) solution as the mobile phase, column temperature of 35 °C, flow rate of 1.0 mL/min; ELSD parameters: nebulization temperature of 50 °C, drift tube temperature of 110 °C, nitrogen flow rate of 2.2 mL/min, gain value of 1.0. During the natural dormancy of C.pilosula, trehalose was only detected in roots (pre-dormant) and buds (dormant), raffinose was only detected in pre-dormant roots; fructose and glucose were found in higher levels in the leaves in the early stage of dormancy, and sucrose was found in higher levels in the roots in the early stage of dormancy and the buds in the dormant period. The highest levels of all three types of sugars were found in the buds after dormancy was released. This study established an efficient HPLC-ELSD technical system for the simultaneous detection of five soluble sugars, and confirmed that trehalose and raffinose are the main soluble sugar components of C.pilosula for cold resistance. Trehalose is synthesized in the roots and further transported to the buds. Fructose, glucose and sucrose are soluble sugar components stored in the roots of C.pilosula, and are also the source of substances and energy for germination.

Key words: Codonopsis pilosula (Franch.) Nannf., Soluble sugar component, HPLC-ELSD, Dormancy, Stress physiology

Fig.1

Chromatogram of soluble sugar in different volume ratios of acetonitrile and water 1: Fructose; 2: Glucose; 3: Sucrose; 4: Trehalose; 5: Raffinose. The same below."

Fig.2

Chromatogram of soluble sugar at different drift tube temperatures"

Table 1

Regression equation, correlation coefficient and linearity range for the five soluble sugars"

组分Composition 回归方程Regression equation 相关系数Correlation coefficient 线性范围Linearity range (mg/mL)
果糖Fructose y=2055.4x-31.834 0.9998 0.016~10
葡萄糖Glucose y=2326.7x-5.2971 0.9998 0.016~10
蔗糖Sucrose y=2530.5x+120.21 0.9991 0.016~10
海藻糖Trehalose y=2623.6x-57.838 1.0000 0.016~10
棉子糖Raffinose y=2250.1x-256.03 0.9990 0.016~10

Fig.3

Chromatogram of standard and Codonopsis pilosula samples (a) Standard sample; (b) Predormancy root; (c) Predormancy stem; (d) Predormancy leaf; (e) Predormancy bud; (f) Dormant bud; (g) Bud after release from dormancy."

Fig.4

Contents of the five soluble sugars in each organ during the pre-dormant period Different lowercase letters show significant differences (P < 0.05), the same below."

Fig.5

Contents of five soluble sugars in buds in different dormancy periods"

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