|
| ORIGINAL ARTICLE |
|
| Year : 2010 | Volume
: 6
| Issue : 24 | Page : 259-263 |
 |
|
Determination of total flavonoids in three Sedum crude drugs by UV-Vis spectrophotometry
Yujie Chen, Jing Wang, Dingrong Wan
Pharmacy College of South-Central University for Nationalities, Wuhan, Hubei Province - 430074, People's Republic China
| Date of Submission | 11-Jun-2010 |
| Date of Decision | 22-Jul-2010 |
| Date of Web Publication | 20-Oct-2010 |
Correspondence Address:
Dingrong Wan
Pharmacy College of South-Central University for Nationalities, Wuhan, Hubei Province - 430074
People's Republic China
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/0973-1296.71784
 Abstract | | |
A simple, rapid UV-Vis spectrophotometry method for the determination of total flavonoids in Sedum sarmentosum Bunge., S. lineare Thunb., and S. erythrostictum Migo. was developed, with a good linearity, precision, and stability. The detection wavelength was set at 500 nm, and an extraction solvent was optimized. Through the comparative study of multiple samples of the three plant drugs, their collected seasons and the habitats can be preliminarily ascertained, which may help to control the quality of the medicines to some extent. Keywords: Genus Sedum, quality control, total flavonoid content, UV-Vis spectrophotometry
How to cite this article:
Chen Y, Wang J, Wan D. Determination of total flavonoids in three Sedum crude drugs by UV-Vis spectrophotometry. Phcog Mag 2010;6:259-63 |
How to cite this URL:
Chen Y, Wang J, Wan D. Determination of total flavonoids in three Sedum crude drugs by UV-Vis spectrophotometry. Phcog Mag [serial online] 2010 [cited 2022 Jul 4];6:259-63. Available from: http://www.phcog.com/text.asp?2010/6/24/259/71784 |
| Introduction | |  |
Sedum sarmentosum Bunge., S. lineare Thunb., and S. erythrostictum Migo. are three Sedum plant medicines widely used against hepatitis, dysentery, herpes zoster, and swelling in China. Since S. sarmentosum Bunge. is famous for its validity of acute or chronic hepatitis, it has been recorded by Chinese Pharmacopoeia (2010), and various preparations have been developed from it. According to the present reports, the active components of S. sarmentosum Bunge. due to liver protection and alanine aminotransferase (ALT) decrease mainly exist in the water portion and n-butanol portion which principally include the glycosides and flavonoids, suggesting that the medicinal properties of S. sarmentosum Bunge. might result from pharmacologically active bioflavonoids. [1],[2],[3],[4],[5] Thus, the total flavonoid determination could reflect the quality of the drug to some extent.
Traditional extraction techniques, such as maceration, heating reflux, and soxhlet extraction, are often effective but time consuming or labor intensive. In contrast, ultrasonic extraction can extract analytes from various matrices in a shorter time. Besides, the quantification of total flavonoids was usually completed by UV-Vis absorption at 500 nm and calculated against a stand reference in routine analyses. [6],[7],[8],[9] Although a number of papers have reported on the quantification of total flavonoids in traditional herbal medicines, few described the determination combined with comparison among several kinds of herbal drugs from different habitats as well as various months of collection together.
This article aimed to develop a rapid simple approach to estimate the quality of the three crude medicines and try to preliminarily explore the relationships among the harvest period, habitat, and drug quality.
| Materials and Methords | | |
Apparatus
KQ-500E ultrasonic apparatus (Kunshan Ultrasonic Instrument Co., Ltd., Beijing, People's Republic of China) was applied here and the outpower was 500 W, with a frequency of 40 kHz. The determination was performed on a 757 CRT UV-Vis spectrophotometer (Lengguang Instrument Co., Ltd., Shanghai, People's Republic of China).
Reagents and materials
The standard rutin was purchased from National Institute for the Control of Pharmaceutical and Biological Products (batch number: 100080-200707); the ethanol and petroleum ether employed were of the analytical reagent grade. Double distilled water was used in this work.
The three plant medicines, harvested from Jianshi, Yichang, Huangmei, and Wuhan, Hubei province, in different months were air-dried immediately under 60°C and were respectively identified as S. sarmentosum Bunge., S. lineare Thunb., and S. erythrostictum Migo. by Professor Wan Dingrong from College of Pharmacy, South-Central University for Nationalities [Table 1].
Sample preparation
Each dried plant drug was crushed and passed through a 20-mesh sieve. Then 1.0 g of each sample was accurately taken, mixed with 50 ml of petroleum ether, and extracted ultrasonically twice for 40 min. Poured out the petroleum ether solution. After the sample powder was dried completely, 100 ml of 70% ethanol was added, and subjected to another ultrasonic extraction for 90 min (3 Χ 30 min) at 55°C, and then filtered into a flask to make a total volume of 100 ml with 70% ethanol.
Standard preparation
A total of 200.0 mg/l of the rutin standard solution was prepared by dissolving rutin reference material in 70% ethanol.
Procedures for the determination of total flavonoids
A total of 4 ml of each sample extraction was pipetted into a 10-ml volumetric flask. The solution was treated with 0.40 ml of the 5% NaNO 2 solution for 6 min and evenly mixed, into which 0.4 ml of the 10% Al(NO 3 ) 3 solution was added and shaked up; then 6 min later, 4ml of the 4% NaOH solution was added to it. The mixture was diluted to the volume with double distilled water, and allowed to stand for 15 min before analyzing against the blank solution.
| Results and Discussion | | |
Selection of the detection wavelength
The absorption spectra of the three sample extraction and rutin solutions were obtained. The absorption peak of rutin was at 510 nm and S. lineare Thunb. at 500nm, while both S. lineare Thunb. and S. erythrostictum Migo. solutions had strong absorptions at a little less than 500 nm. Comprehensively, 500 nm was chosen as the detection wavelength.
Optimization of the extraction solvent
Since flavonoids contain some hydroxyl groups, they often dissolve easily in alcohol or alcohol-water mixtures. The S. sarmentosum Bunge. sample (Jianshi, June 28, 2006) was employed here, trying to find out the optimum solvent for sample extraction. The relationship between alcohol concentration and the yield of the total flavonoids is shown in [Figure 1]. In the figure, we could see that the extraction yield kept increasing and reached the top when the ethanol concentration went up to 70%, and then began to fall sharply as the ethanol concentration continued growing. Hence, the optimum extraction solvent turned out to be 70% ethanol.  | Figure 1: The relationship between the ethanol concentration and the flavonoid extraction yield of S. sarmentosum Bunge.
Click here to view |
Method validation
All the sample solutions applied here were made from the S. sarmentosum Bunge. sample collected in Jianshi, June 28, 2006.
The linearity of the method was tested by analyzing different amounts of the standard solution (0, 0.2, 0.5, 1, 2, 3, 4 ml, respectively) coupled with isometric color development reagents according to the procedures for the determination of total flavonids. A good linear response was shown (r > 0.999) over the range of 4-80 μg/ml, with an equation A = 12.155C + 0.005, in which A meant the absorbance value while C (mg/mL) represented the concentration of the rutin solution.
Exactly took 4 ml of the S. sarmentosum Bunge. sample solution, operated according to the colorimetric method mentioned above and determined successively for five times. The average absorption was 0.489, and the RSD attained was 0.09%, demonstrating that the instrument used had a high precision.
A series of S. sarmentosum Bunge. samples were taken and treated according to sample preparation and colorimetric methods mentioned above. The flavonoid contents calculated from regression equation were 1.06%, 1.05%, 1.01%, 1.04%, 1.01%, and 1.00%, so the average content was 1.03% with RSD < 2.7% (n = 6), investigating a good repeatability.
Recovery experiment was performed to evaluate the accuracy of the methods. 1.0g of S. sarmentosum Bunge. sample powder were spiked with 0.5ml rutin solution (containing 10 mg of rutin) prior to the extraction. The spiked samples were analyzed in six copies. Recoveries of total flavonoids obtained are shown in [Table 2], which informed us that the method possessed a nice accuracy.
After the extraction of S. sarmentosum Bunge., the sample mentioned above was dealt with according to the procedures for the determination of total flavonoids; absorptions were measured every 20 min ranging from 0 min to 120 min [Figure 2]. It's clear that the absorption of the solution was relatively stable (RSD = 0.78%) if the measurement was carried out within 120 min. Thus, all the analyses should be performed within 2 h after the color reaction.
Sample analysis
The present method was applied to the analyses of flavonoid contents in three crude drugs from Genus Sedum and the results are displayed in [Table 3]. It is obvious that all the related plant drugs contain flavonoids, which may be associated with their common pharmacological actions.
A relationship between total flavonoid content and harvest season is also revealed by [Table 3]. All the three plant medicines contained maximum flavonoids in April (flowering period), and then the flavonoid quantities kept decreasing from April to June or July (during this time, the leaves of S. erythrostictum Migo. mostly fell down). But for S. sarmentosum Bunge. and S. lineare Thunb., the contents began to rise a little after August [Figure 3] and [Figure 4] (except S. sarmentosum Bunge. sample collected in Wuhan), while the flavonoid quantities of S. erythrostictum Migo. continued to drop during autumn [Figure 5]. It seemed that all the three plant drugs should have the best quality if harvested in flowering time. Thus, the quality could be preliminarily controlled by flavonoid determination.  | Figure 3: The relationship among month of collection, habitat and the content of total flavonoids in S. sarmentosum Bunge.
Click here to view |
 | Figure 4: The relationship between month of collection and the content of total flavnoids in S. lineare Thunb
Click here to view |
 | Figure 5: The relationship among month of collection, habitat and the content of total flavnoids in S. erythrostictum Migo
Click here to view |
On the other hand, it was also manifested that a certain relation existed between the flavonoid content and the sample habitat. Both [Figure 3] and [Figure 5] informed us that samples collected in the west of Hubei Province (Jianshi or Yichang) usually possessed higher flavonoid quantities than those collected in the east. Therefore, it was reasonable to deduce that Western Hubei should be a more suitable habitat to collect the three crude drugs from genus Sedum than the east.
| Conclusion | | |
As the components contained in a Chinese traditional medicine are various and complicated, the strong polar glycosides, i.e., S. sarmentosum Bunge. glycoside (Sarmentosin), are not the only active composition associated with hepatitis treatment. Flavonoid ingredients are also the key point. Therefore, the flavonoid quantities reflected the quality of the three medicines from one aspect.
Studies on total flavonoid determination of the S. sarmentosum Bunge., S. lineare Thunb., and S. erythrostictum Migo. samples collected from different habitats and months clearly manifested that the flavonoid contents are closely related to the collected seasons, which may help to judge the optimum harvest time of the three plant drugs, so that the quality could be preliminarily controlled. The results also suggested that the flavonoid content changed with the habitat. The samples harvested in the west of Hubei usually possessed higher flavonoid contents compared with those collected in the east.
The method established for the flavonoid determination in three crude drugs of genus Sedum was simple, direct, and accurate, providing a valuable reference for quality control.
| References | | |
| 1. | Wan DR, Chen JC, Yu HH. Records on Material Medical of Hubei. Vol. 1. Wuhan: Hubei Science and Technology Press; 2002. p. 319.  |
| 2. | Pan JH, He MT. The screening of the components due to protect liver and decrease the ALT in Sedum erythrostictum Migo. Chin Pharm Aff 2002;16:365-6. |
| 3. | He AM, Wang MS. The flavonoids in Sedum erythrostictum Migo. Chinese Trad Herbal Drugs 1997;28:517-22. |
| 4. | Pan JH, He MT, Luo L, Yan J. Test of protecting liver and reducing enzyme levels on different extraction sites from sedum sarmentosum bunge. Lishizhen Med Mater Med Res 2001;12:888. |
| 5. | Pan JH, Xue CY, Yan J, Liu XH. Test of protecting liver and reducing enzyme levels of isorhamnetin-3,7-di-O-β-D-glucopyranoside. Lishizhen Med Med Mater Res 2002;13:714-5. |
| 6. | Kartashova GS, Sudos LV. Quantitative determination of flavonoids in the above-ground part of agrimony plants. Pharm Chem J 1997;31:488-91. |
| 7. | Heimler D, Pieroni A, Tattini M, Cimato A. Determination of flavonoids, flavonoid glycosides and biflavonoids in Olea europaea L. Leaves. Chromatographia 1993;33:369-73. |
| 8. | Marinova D, Ribarova F, Atanassova M. Total phenolics and total flavonoids in bulgarian fruits fruits and vegetables. J Univ Chem Technol Metallurgy 2005;40:255-60. |
| 9. | Soares LA, Bassani VL, Ortega GG, Petrovick PR. Total flavonoid determination for the quality control of aqueous extractives from Phyllanthus niruri L. Lat. Am J Pharm 2003;22:203-7. |
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3]
| This article has been cited by | | 1 |
A DFT study of the chemical reactivity properties, spectroscopy and bioactivity scores of bioactive flavonols |
|
| Eric O. Akintemi, Krishna K. Govender, Thishana Singh | | Computational and Theoretical Chemistry. 2022; 1210: 113658 | | [Pubmed] | [DOI] | | | 2 |
Synergistic mechanism for the bioactivity fortification of licorice by honey |
|
| Shasha Kong, Pengyue Li, Robert Verpoorte, Jia Wang, Chao Zhu, Yuntao Dai, Shilin Chen | | Journal of Ethnopharmacology. 2022; : 115048 | | [Pubmed] | [DOI] | | | 3 |
A Highly Sensitive Electrochemical Sensor Based on ß-cyclodextrin Functionalized Multi-Wall Carbon Nanotubes and Fe3O4 Nanoparticles for Rutin Detection |
|
| Wen-Jun Zhao, Bao-Lin Xiao, Xin-Yan Song, Xin Meng, Xin-Xin Ma, Yang-Yang Li, Jun Hong, Ali Akbar Moosavi-Movahedi | | Journal of The Electrochemical Society. 2022; 169(4): 047509 | | [Pubmed] | [DOI] | | | 4 |
Sedum sarmentosum Bunge Attenuates Drug-Induced Liver Injury via Nrf2 Signaling Pathway: An Experimental Verification Based on Network Pharmacology Prediction |
|
| Zhitao Jiang, Yi Han, Yuechan Zhang, Jie Li, Chundi Liu, Malik Alazzam | | Journal of Healthcare Engineering. 2021; 2021: 1 | | [Pubmed] | [DOI] | | | 5 |
Occurrence of Grey Leaf Spot ofSedum erythrostictumCaused byCercosporacf.pseudokalanchoesin China |
|
| Yue Yuan,Rujun Zhou,Haijiao Xu,Junfan Fu,Ning Hao,Hongbo Li | | Journal of Phytopathology. 2015; 163(11-12): 997 | | [Pubmed] | [DOI] | | | 6 |
Enhancement of flavonoids extraction from fig leaf using steam explosion |
|
| Lanzhi Qin,Hongzhang Chen | | Industrial Crops and Products. 2015; 69: 1 | | [Pubmed] | [DOI] | | | 7 |
A new, simple colorimetric method for assaying diosmin, and flavonoids in daflon tablets and orange peel extracts |
|
| Mir, S.A. and Ahangar, A.A. and Bhat, A.S. | | International Journal of PharmTech Research. 2013; 5(2): 341-348 | | [Pubmed] | | | 8 |
Spectrophotometric assays for flavonoids diosmin, quercetin, rutin and morin with copper, molybdenum, lead and tungsten |
|
| Mir, S.A. and Ahangar, A.A. and Bhat, A.S. | | International Journal of PharmTech Research. 2013; 5(2): 383-390 | | [Pubmed] | | | 9 |
Enhanced water-solubility of Licorice extract microparticle prepared by antisolvent precipitation process |
|
| Chang Zu,Xiuhua Zhao,Xiaowei Du | | Advanced Powder Technology. 2013; | | [Pubmed] | [DOI] | | | 10 |
Determination of quercetin, plumbagin and total flavonoids in Drosera peltata Smith var. glabrata Y.Z.Ruan |
|
| He, Y. and He, Z. and He, F. and Wan, H. | | Pharmacognosy Magazine. 2012; 8(32): 263-267 | | [Pubmed] | | | 11 |
Microwave-assisted extraction and antihyperlipidemic effect of total flavonoids from corn silk |
|
| Yan, Z. and da-Yun, S. and Jing-Shu, Z. and Hong-Li, Z. | | African Journal of Biotechnology. 2011; 10(65): 14583-14586 | | [Pubmed] | | | 12 |
In-vitro evaluation of bioactive compounds, anti-oxidant, lipid peroxidation and lipoxygenase inhibitory potential of Citrus karna L. peel extract |
|
| Jagdeep Singh,Shailja Sood,Arunachalam Muthuraman | | Journal of Food Science and Technology. 2011; | | [Pubmed] | [DOI] | |
|
 |
|
|
|
