3,5-二硝基水杨酸法的优化及日本落叶松木质部发育中阿拉伯半乳聚糖的含量变异

doi:10.11707/j.1001-7488.LYKX20230597

Abstract

Abstract:

Objective: This study aims to establish the optimal detection and hydrolysis conditions for the 3,5-dinitrosalicylic acid method, in order to accurately determine the content and its variation of arabinogalactan in the xylem of Larix kaempferi, and provide technical support for deeply understanding the distribution, development, and biosynthesis of arabinogalactan in the xylem of L. kaempferi. Method: The optimal detection conditions of 3,5-dinitrosalicylic acid method were determined by analyzing the three aspects of coloration reagent volume, reaction time, and reaction temperature. At the same time, an orthogonal test was used to optimize the hydrolysis conditions of total sugar from three aspects of hydrochloric acid concentration, hydrochloric acid volume and hydrolysis time. The contents of total sugar and reducing sugar were measured, respectively, with which the polysaccharide content was calculated. Result: The optimum detection conditions of the 3,5-dinitrosalicylic acid method were established. The coloration reagent volume, reaction time, and reaction temperature were 5 mL, 9 min, and 90 ℃, respectively. The optimal hydrolysis conditions were hydrochloric acid concentration of 6 mol·L^?1, hydrochloric acid volume of 15 mL, and hydrolysis time of 40 min. In individual L. kaempferi trees, the content of arabinogalactan in xylem gradually decreased with the increase of tree height in the axial direction, and was much lower in the sapwood than that in the heartwood in the radial direction. Overall, the arabinogalactan content in heartwood area gradually increased outward with the growth ring, while the arabinogalactan content in the sapwood area gradually increased inward the growth ring. The arabinogalactan content was highest in the transition zone between sapwood and heartwood, the arabinogalactan content in springwood was much higher than that in summerwood within each growth ring, and the arabinogalactan content in earlywood was much higher than that in latewood within the transition zone. Conclusion: In this study, a simple and stable method for accurately detecting the content of arabinogalactan without relying on high-quality samples has been established. At the same time, it has been found that arabinogalactan content is highest in the basal trunk, and is higher in the heartwood than in the sapwood, and higher in the springwood than in the summerwood, and highest in the transition zone, suggesting that the arabinogalactan may be synthesized in the transition zone and related to the formation of heartwood.

Key words: Larix kaempferi, arabinogalactan, 3,5-dinitrosalicylic acid method, content variation

CLC Number:

S718.43

Kaikai Zhang,Yunhui Xie,Xiaomei Sun. Optimization of 3,5-Dinitrosalicylic Acid Method and Variation of Arabinogalactan Content in the Developing Xylem of Larix kaempferi[J]. Scientia Silvae Sinicae, 2025, 61(2): 113-121.

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试验号 No.	因素 Factors			方案 Programs
试验号 No.	A 水解时间 Hydrolyzing time/min	B 盐酸浓度 HCl concentration/ (mol·L^?1)	C 盐酸体积 HCl volume/ mL	方案 Programs
1	20 (1)	6 (1)	5 (1)	A1B1C1
2	20 (1)	9 (2)	10 (2)	A1B2C2
3	20 (1)	12 (3)	15 (3)	A1B3C3
4	30 (2)	6 (1)	10 (2)	A2B1C2
5	30 (2)	9 (2)	15 (3)	A2B2C3
6	30 (2)	12 (3)	5 (1)	A2B3C1
7	40 (3)	6 (1)	15 (3)	A3B1C3
8	40 (3)	9 (2)	5 (1)	A3B2C1
9	40 (3)	12 (3)	10 (2)	A3B3C2

试验号 No.	因素 Factors			方案 Program	吸光值 Absorbance
试验号 No.	A 水解时间 Hydrolyzing time/min	B 盐酸浓度 HCl concentration/ (mol·L^?1)	C 盐酸体积 HCl volume/mL	方案 Program	吸光值 Absorbance
1	20	6	5	A1B1C1	0.004 0
2	20	9	10	A1B2C2	0.000 3
3	20	12	15	A1B3C3	0.017 3
4	30	6	10	A2B1C2	0.014 7
5	30	9	15	A2B2C3	0.003 7
6	30	12	5	A2B3C1	0.008 3
7	40	6	15	A3B1C3	0.025 0
8	40	9	5	A3B2C1	0.020 3
9	40	12	10	A3B3C2	0.017 7
k₁	0.007 2	0.014 6	0.010 9
k₂	0.008 9	0.008 1	0.010 9
k₃	0.021 0	0.014 4	0.015 3
R	0.013 8	0.006 4	0.004 4

项目Item	1	2	3	4	5	6	平均值 Mean	相对标准偏差 RSD (%)
总糖Total sugar	0.198	0.198	0.199	0.199	0.199	0.198	0.199	0.252
还原糖Reducing sugar	0.115	0.115	0.116	0.115	0.116	0.116	0.116	0.433

项目Item	样本1 Sample 1	样本2 Sample 2	样本3 Sample 3	样本4 Sample 4	相对标准偏差 RSD (%)
总糖Total sugar	0.134	0.123	0.127	0.128	2.917
还原糖Reducing sugar	0.104	0.102	0.099	0.100	1.821

项目Item	时间 Time/min										相对标准偏差 RSD (%)
项目Item	0	10	20	30	40	50	60	70	80	90	相对标准偏差 RSD (%)
总糖Total sugar	0.199	0.203	0.212	0.200	0.208	0.207	0.208	0.207	0.206	0.208	1.763
还原糖Reducing sugar	0.155	0.119	0.119	0.117	0.120	0120	0.119	0.119	0.120	0.119	1.280

Optimization of 3,5-Dinitrosalicylic Acid Method and Variation of Arabinogalactan Content in the Developing Xylem of Larix kaempferi

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日本落叶松还原糖浓度 Concentration of reducing sugar in Larix kaempferi/(mg·mL^?1)	加标浓度 Spiked content/(mg·mL^?1)	理论值 Theoretical value/(mg·mL^?1)	实测值 Actual measurement value/(mg·mL^?1)	回收率 Rate of recovery (%)	相对标准偏差 RSD (%)
0.181	1.500	1.714	1.790	105.047	0.417
0.181	1.500	1.714	1.793	105.225
0.181	1.500	1.714	1.805	106.058
0.181	2.000	2.214	2.266	102.567	0.585
0.181	2.000	2.214	2.275	103.034
0.181	2.000	2.214	2.295	104.017
0.181	2.500	2.714	2.607	95.715	0.143
0.181	2.500	2.714	2.615	96.022
0.181	2.500	2.714	2.608	95.748

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