Analysis Of Bioactive Substances And Origin Differences Of Cistanche Tubulosa in Xinjiang ②
Dec 15, 2022
2 Results and Analysis
2.1 Analysis of bioactive substances in Cistanche tubulosa from different producing areas
Figure 2 shows the content of biologically active substances in Cistanche tubulosa from different origins, where different lowercase letters indicate significant differences between groups (P<0. 05), the same below. It can be seen from Figure 2a) that the total polyphenol content in the roots of the samples from A, B, C, and D was significantly higher than that in the middle and top; in the samples from E and F, the total polyphenol content in the top was higher. Among the samples from different origins, A1 (28. 32 mg GAE/ g DW) had the highest total polyphenol content, followed by D1 (26. 41 mg GAE / g DW) and F3 ( 25. 69 mg GAE / g DW); the lowest total polyphenol content was A3 ( 7. 45 mg GAE / gDW). The order of the average values of the total polyphenol content in the three parts of the samples from each origin is: F ( 23. 57 mg GAE / g DW) > D ( 19. 44 mg GAE / g DW) > E (19. 17 mg GAE / g DW) >C(18. 67 mgGAE/gDW)>B(18. 41 mg GAE/gDW)>A(16. 87 mgGAE/gDW). It can be seen from Figure 2b) that in the samples from the four origins of A, B, C, and D, the total flavonoid content in the top was significantly higher than that in the middle and roots. Among the samples from different origins, A3 (14. 66 mg RE / g DW) had the highest total flavonoid content, followed by A2 (13. 48 mg RE / g DW). The order of the average values of total flavonoids content in 3 parts of samples from each origin is: A(13. 15 mg RE / gDW) >E(11. 71 mg RE / g DW) >F(6. 53 mg RE / gDW) > B ( 4. 88 mg RE / g DW) > C ( 4. 83 mg RE / g DW) > D (3. 93 mg RE / g DW).
It can be seen from Figure 2c) that the samples from the four origins of A, B, D, and E had higher total triterpene content in the roots; the samples from the two origins of C and F had higher total triterpene contents in the top. Among the samples from different origins, F3 (14. 43 mg OAE/g DW) and B1 (13. 78 mg OAE/g DW) had the highest content of total triterpenes.
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The order of the average content of total triterpenes in the three parts of the samples from each origin is: F(13. 54 mg OAE/ g DW)>A(13. 01 mg OAE/ g DW)>B(11. 26 mg OAE/ g DW) >C(10.97 mg OAE/g DW)>E(9.77 mg OAE/g DW)>D(9.30 mg OAE/g DW).
It can be seen from Figure 2d) that the total polysaccharide content in the roots of samples from A, B, D, and E was significantly higher than that in other parts, which is consistent with the research results of Yang Taixin et al. [25]. Among the samples from different origins, A1 (32. 34 mg DE / g DW) had the highest total polysaccharide content, followed by E1 (26. 52 mg DE / g DW) and B1 (25. 78 mg DE / g DW); The lowest content is D3 (7. 24 mg DE / g DW). In samples from C and F, the total polysaccharide content of C3 (25. 4 mg DE / g DW) and F3 (24. 24 mg DE / gDW) was higher. The average order of the total polysaccharide content of the three parts of the samples from each origin is: A (20. 41 mg DE / g DW) > C (18. 75 mg DE / g DW) > E (18. 57 mg DE / g DW) >F (17. 53 mg DE / g DW) > B (16. 91 mg DE / g DW) > D (10. 07 mg DE / g DW). It can be seen from Figure 2e) that except for origin A and B, the top proanthocyanidin content in samples from the other four origins was higher than that in other parts, among which C3 (5. 56 mg PCE / g DW), D3 (5. 73 mg PCE / g DW) g DW) and F3 (5. 93 mg PCE / g DW) had higher proanthocyanidin content. The order of the mean values of proanthocyanidin contents in three parts of samples from each origin was: F(4. 86 mg PCE / g DW)>D(4. 70 mg PCE / g DW)>C(4. 00 mg PCE / g DW)> A(3.99 mg PCE/g DW)>B(3.37 mg PCE/g DW)>E(3.28 mg PCE/g DW).
From Figure 2f), it can be seen that in the samples from the three production areas of A, B, and E, the content of echinacoside in the roots is relatively high, and the content of B1 reaches 41.72 mg / gDW; in the samples from the three production areas of C, D, and F, the echinacoside content in the top The content of inulin was higher, and the content of C3 was as high as 32. 88 mg / g DW, followed by F3 (27. 26 mg / g DW). The order of the mean values of echinacoside content in three parts of the samples from each origin is: B ( 31. 63 mg / g DW ) > F ( 22. 52 mg / g DW ) > C ( 20. 84 mg / g DW ) > A
( 18. 65 mg / g DW ) > E ( 14. 46 mg / g DW ) > D (3. 97 mg / g DW ). It can be seen from Fig. 2g) that in the samples from A and E, the root acteoside content is relatively high; in the samples from B, C, D, and F, the top verbascoside content is high, and the highest content is B3 ( 9. 32 mg / g DW), followed by F3 (8. 47 mg / gDW) and C3 (7. 84 mg / g DW). The order of the mean values of verbascoside content in three parts of the samples from each origin was: B ( 7. 91 mg / g DW) > F (6. 12 mg / g DW) > C (5. 39 mg / g DW) > A (4 .92 mg/g DW)>E(4. 12 mg/g DW)>D(1. 65 mg/gDW). Guo Xiongfei et al[1,15] studied the contents of echinacoside and verbascoside in different parts of Cistanche tubulosa, and the results showed that the contents of echinacoside and verbascoside in the roots were significantly higher than those in other parts; Wang Guoping[26] found that, The content of echinacoside in Cistanche from different origins in southern Xinjiang is 0. 35% ~11. 93%. In this study, the contents of total polyphenols, total polysaccharides and total triterpenes in the roots of most samples were relatively high, while the contents of individual components in the roots of individual samples were relatively low. The specific reasons need further study.
In summary, in different parts of most samples, the contents of total polyphenols, total triterpenes, and total polysaccharides in the roots are higher, and the contents of total flavonoids and total proanthocyanidins in the top are higher; The content of glycosides and echinacoside was the highest, and the content of total flavonoids and total triterpenoids of samples from A producing area was higher.
2.2 Antioxidant activity analysis of Cistanche tubulosa from different origins
Different bioactive substances in plants may exhibit synergistic or antagonistic effects, so the determination of the antioxidant activity of plant sample extracts is more representative than the determination of the antioxidant activity of a single substance[27]
. Figure 3 shows the antioxidant activity of Cistanche from different origins. It can be seen from Figure 3a) that the scavenging ability of samples from different origins to DPPH free radicals is significantly different. The samples with strong antioxidant activity are A1, B3, C3, D1, E1 and F1, and their IC50 DPPH are 0. 76 mg / mL, 0. 55 mg / mL, 0. 14 mg / mL, 0. 57 mg, respectively / mL, 1. 38 mg / mL and 0. 69 mg / mL. The average values of IC50 DPPH of the 3 parts of the samples from each origin were A (3. 14 mg / mL), B (1. 40 mg / mL), C (1. 12 mg / mL), D (3. 00 mg / mL), respectively. mL), E (3. 61 mg / mL) and F (1. 05 mg / mL), all less than 7. 00 mg / mL. F origin sample
The scavenging ability of the samples from the origin of E to DPPH free radicals is the strongest, and the scavenging ability of the samples from E origin is the weakest.
Figure 3 Antioxidant activity of Cistanche in different places
Fig. 3 Antioxidant activities of C. tubulosa (Schrenk)
Wight from different origins
It can be seen from Figure 3b) that the scavenging ability of samples from different origins to ABTS free radicals is significantly different. The samples with strong antioxidant activity were A1, B1, C3, D1, E1 and F3, and their IC50 ABTS were 1. 07 mg / mL, 0. 91 mg / mL, 0. 81 mg / mL, 1. 93 mg / mL, 1. 41 mg / mL and 1. 23 mg / mL. The IC50 ABTS average values of the 3 parts of the samples from each origin were A ( 1. 88 mg / mL ), B ( 1. 16 mg / mL ), C ( 1. 19 mg / mL ), D ( 2. 33 mg / mL ) ), E ( 1. 49 mg / mL ) and F ( 1. 31 mg / mL ), all less than 3. 00 mg / mL. The scavenging ability of samples from B origin to ABTS free radicals
The ability to scavenge ABTS free radicals was the strongest, and the sample from origin D had the weakest ability to scavenge ABTS free radicals. Bao Bin et al[27] found that the IC50DPPH of methanol and ethanol extracts of Cistanche deserticola were 0. 142 mg/mL and 0. 175 mg/mL, respectively, He Mengmeng et al[12] found that the water extract of Cistanche deserticola DPPH and
The IC50 of ABTS free radical scavenging rate is 0. 225 mg / mL and 1. 819 mg / mL, which is consistent with the results of this study, showing that Cistanche tubulosa has strong antioxidant activity.
2.3 Correlation analysis
Through the Pearson correlation analysis, the correlation among the quality indicators of Cistanche tubulosa from different origins was statistically analyzed, and the results are shown in Table 2. It can be seen from Table 2 that the total polyphenol content was significantly positively correlated with the total polysaccharide content (P<0.05); the total triterpenoid content was significantly positively correlated with the total polysaccharide, echinacoside and verbascoside content (P<0. 05); the total polysaccharide content was significantly positively correlated with the echinacoside content (P<0. 05); the echinacoside content was extremely significantly positively correlated with the verbascoside content (P <0. 01); the IC50 DPPH was significantly correlated with the total The contents of polyphenols and echinacoside were significantly negatively correlated (P<0.05), indicating that the scavenging ability of Cistanche tubulosum on DPPH free radicals was closely related to the contents of total polyphenols and echinacoside; IC50 ABTS and total polyphenols , total polysaccharides, echinacoside and verbascoside
The content was extremely significantly negatively correlated (P<0. 01), and was extremely significantly positively correlated with IC50 DPPH (P<0. 01), indicating that the scavenging ability of Cistanche tubulosa on ABTS free radicals was closely related to total polyphenols, total polysaccharides, pine The contents of guavacoside and verbascoside were closely related.
2.4 Principal component analysis and comprehensive quality evaluation
Performing principal component analysis on various indicators of Cistanche tuberosa, and extracting principal components according to the eigenvalues and contribution rates of each principal component, can improve the efficiency and reliability of the quality evaluation of Cistanche tubulosa[28]. The variance contribution rate of each principal component is shown in table 3. It can be seen from Table 3 that the eigenvalues of the extracted three principal components are all >1, and the cumulative contribution rate of variance is 80.57%, indicating that the extracted three principal components can effectively represent most of the information of all indicators, and comprehensively reflect the quality characteristics.
The principal component factor is rotated, and its load value can reflect the relationship between the principal component factor and various indicators. The results are shown in Figure 4 and Table 4. It can be seen from Figure 4 and Table 4 that PC1 is mainly related to the content of total triterpenoids, total polysaccharides, echinacoside and verbascoside, PC2 is mainly related to the content of total polyphenols and total flavonoids, and PC3 is mainly related to the content of total proanthocyanidins. Taking the variance contribution rate of the three principal components as weights, the following comprehensive evaluation model for the quality of Cistanche tubulosa was constructed: comprehensive evaluation index = 46. 24% F1 + 19. 81% F2 + 14. 52% F3 Through this model, the quality of each production area can be calculated. Sample scores (see Table 5). The seven quality factors of 18 samples were further reduced into three common factors by principal component analysis, which contained 80.57% of the original information. The variance contribution rate of each principal component factor was used as the weight to substitute into the above comprehensive evaluation model, and the quality of Cistanche tubulosa was comprehensively evaluated.
The higher the score, the better the quality of Cistanche tubulosa[29-30]. Based on the average value of the root, middle and top comprehensive evaluation indexes of samples from different places of origin, the quality of Cistanche tuberosa from different places of origin was sorted.
A(0. 445)>F(0. 313)>B(0. 051)>E(-0. 057)>C(-0. 071)>D(-0. 680).
2.5 Cluster analysis
Based on the average quality indexes of the three parts of Cistanche tubulosa from different origins, cluster analysis was performed on samples from different origins [31], and the results are shown in Figure 5. It can be seen from Figure 5 that when the distance is 12.5, 6 production areas can be clustered into 3 categories, and the first category gathers 4 production areas, namely A, E, C and F.
Type 2 and Type 3 each gather one place of origin, B and D respectively. Table 6 shows the mean values of each measurement index in different clusters. It can be seen from Table 6 that the average content of total polyphenols, total flavonoids, total triterpenes and total polysaccharides in group 1 is the highest; the average content of echinacoside and verbascoside in group 2 is the highest, the average content of total polyphenols and total proanthocyanidins is the lowest, and It has the strongest scavenging ability on DPPH and ABTS free radicals; the content of total flavonoids, total triterpenes, total polysaccharides, echinacoside and verbascoside in group 3 is lower, and the scavenging ability on DPPH and ABTS free radicals is the weakest.
3 Conclusion
In this paper, the biological active substances and antioxidant activities of Cistanche tuberosa from different origins in the Taklamakan Desert in Xinjiang, my country were compared and analyzed.
Table 2 Correlation analysis results of different quality indexes of C. tubulosa (Schreak) Wight from different origins
The results showed that the quality and antioxidant activity of samples from different origins and different sample parts were significantly different.
The production areas with the highest content of total polyphenols, total flavonoids, total triterpenes, total polysaccharides, total proanthocyanidins, echinacoside and verbascoside were: Alar City, Kashi Jiashi County, Alar City, Kashi Jiashi County, Ala Seoul City, Hetian Moyu County, Hetian Moyu County.
The IC50DPPH of each sample was less than 7. 00 mg / mL, and the IC50ABTS was less than 3. 00 mg / mL; the samples with the strongest ability to scavenge ABTS and DPPH free radicals were produced in Alar City and Hetian Moyu County, respectively. Three principal components were extracted by principal component analysis, and the cumulative contribution rate of variance was 80. 57%.
The samples from different origins were divided into three categories by cluster analysis. The samples with the highest comprehensive score were Kashi Jiashi County, followed by Alar City and Hotan Moyu County. The research results of this paper will provide basic data and reference for the quality evaluation, product development and standardization of Cistanche tubulosa in my country.
