Emei different altitude populations of quercetin content in Houttuynia_477

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Emei different altitude populations of quercetin content in Houttuynia_477

    Emei different altitude populations of quercetin content in Houttuynia

     Author: Liu Lei, Wu Wei, Zheng Liang, Sandra Oh

     Abstract Objective To study the Emeishan populations at different altitudes Yuxingcao differences in quercetin

    content. Methods Hypersil ODS C18 (5 μm, 4.0 mm × 250 mm)

    column, mobile phase of acetonitrile: 0.4% phosphoric acid = 45:55, adjusted pH = 3, flow rate 1.0 ml / min, column temperature 30 ?, UV detection wavelength of 370 nm. Results The method is simple, rapid, accurate, and can be used as Yuxingcao quantitative analysis of quercetin. Conclusion of 9 different elevation populations of quercetin Houttuynia there are significant differences in the levels and altitude and the correlation between soil nutrient content are not significant.

     Key words Houttuynia; quercetin; altitude; soil nutrients; RP-HPLC

     Abstract: ObjectiveTo study the difference of quercetin content of H.cordata Thunb. From different altitudes of Mt.Emei.MethodsQuercetin was determined by HPLC on Hypersil ODS C18 (4.0 mm × 250 mm, 5 μm) at 30 ? with acetonitrile:

    0.4% phosphoric acid (45:55), pH = 3 as the mobile phase.Detection wavelengh was 370 nm.The flow rate was 1.0 ml / min.Results This method is simple, fast and accurate, and

    can be the quantitative analytic method of quercetin content of H. cndata.ConclusionThere are significant differences among the quercetin content of 9 populations from different altitudes.The correlationship between quercetin content and

    altitudes and soil nutrient are not significant.

     Key words: Houttuynia cordata Thunb.; Quercetin;

Altitude; Soil nutrient; RP HPLC

     Houttuynia is Saururaceae Houttuynia Houttuynia cordata Thunb. Of the whole plant. Widely distributed in central China, south-east and south-western provinces and autonomous

    regions, especially in Sichuan, Hubei, Hunan, Jiangsu and other provinces mostly. Of Weixin, cold, with heat detoxification, swelling removing the abscess, the diuretic effect of Tonglin. Houttuynia In addition to containing volatile oils, but also contains a large number of quercetin, quercitrin, iso-quercitrin and so on. Quercetin which has expectorant, relieving cough and asthma, anti-inflammatory,

    anti-allergy, expansion of coronary, blood fat, anti-

    arrhythmia, anti-platelet aggregation, anti-oxidation, anti-

    tumor a wide range of pharmacological effects [1], and their contents is a measure of an important indicator of Houttuynia medicinal value.

     Mount Emei is located in the southwest edge of Sichuan Basin, China, was formed in 3000 years ago before the

    Himalayan movement. Mountain for nearly north-south mountain

    ranges, the highest peak altitude 3 099 m. Mount Emei mild climate, clouds and more sunshine less rainfall, air relative humidity, there is a wealth of plant resources. According to

    our previous investigation, Mount Emei Yuxingcao resources are very abundant, mainly in the altitude of 500 ~ 1 800 m range.

     In recent years, the study reported that domestic Yuxingcao mainly in the production of cultivation,

    pharmacological efficacy, clinical applications, and genetic background [2 ~ 5] and so on. Some scholars of some plant secondary metabolites and the relationship between altitude studied [6,7], the majority of results showed that the sub-

    product of chemical and biological Xie and altitude there were significant or very significant correlation. Some academics have also secondary metabolites of plants and soil nutrients in the relationship between the studied [8], results showed that the soil nutrients on plant secondary metabolites of the

    synthesis is also significant. Previously, I had Emeishan populations at different altitudes Houttuynia volatile oil constituents of a research and found that part of the volatile oil composition and altitude, and soil nutrients there were

    significant correlation (ecological Sinica, to be published). This study, using RP-HPLC method were determined at different altitudes 9 Yuxingcao populations quercetin content of altitude and soil nutrients and the relationship between quercetin, through the secondary metabolites and the altitude and soil nutrients in a comprehensive evaluation of artificial cultivation of Houttuynia the appropriate choice of altitude and soil conditions and further rational development and utilization provide some theoretical basis.

     An instrument and reagent

     Agilent's 1100 High Performance Liquid Chromatography (including degasser, quaternary gradient pump, autosampler, column temperature box, DAD detector), Sartorius Corporation BP211D electronic balance, ICE's Micromax centrifuge, Millipope Company MilliQ-type water instrument.

     Chromatography pure acetonitrile by TEDIA companies, quercetin reference substance (purity "98%) from Sigma company, with the remaining reagents of analytical pure.

     Table 1 Test materials, the collection location, altitude, growing season environment and multi-degree


     2 Materials and methods

     2.1 The experimental material due to Emei Mountain for more than 1 800 m above sea level in areas find it difficult to Houttuynia collected, the testing of all populations of the materials are taken from the altitude of 1 800 m below. 9Materials and the growth of populations in soil samples in the20050713Collected from the western slope of Mount Emei. The altitude of populations, growth environment and abundance in Table 1. The identification of Sichuan

    Agricultural University, Dr. Wu Wei are Houttuynia cordata Thunb..

     2.2 The growth of Mount Emei Houttuynia the basic conditions of soil nutrient content in the growth of the Emeishan Yuxingcao soil nutrient content of the basic

    conditions in table 2. Soil sample processing and analysis of the various nutrient elements in the light of "soil agricultural chemistry analysis" in the conventional methods.

     2.3 Chromatographic conditions Column: Hypersil ODS C18 (5 μm, 4.0 mm × 250 mm); mobile phase: acetonitrile: 0.4% phosphoric acid = 45:55, adjusted pH = 3. Flow rate: 1.0 ml / min; column temperature: 30 ?; UV detection wavelength: 370

    nm; quercetin in the control and test samples for the

    chromatographic separation shown in Figure 1.

     Table 2 Growth of Mount Emei Yuxingcao soil nutrient content (abbreviated)

     Data listed in the table 3 times the average of measurement results, "/" indicates not detected due to reasons

    for the test

     Figure 1 Effect of quercetin in control and test samples for the chromatographic separation diagram (omitted)

     2.4 Preparation of standard solution Weigh quercetin precision control, constant volume anhydrous ethanol to 50 ml,

    dubbed the concentration of 224.3 μg / ml of control

    solution, and then diluted dubbed in the concentration of 22.43 μg / ml of control solution.

     2.5 Sample preparation for the test materials will be crushed after drying over 40 mesh sieve, precision Weigh 1 g buy a plug 50 ml flask, add extract (ethanol: 25% hydrochloric acid = 4:1) 30 ml after the ultrasonic extraction 40 min, carried out twice, 20 min / times, the middle interval of 10 min. Remove the filter residue washed several times with the

    extract, constant volume to 50 ml. Taking 1.0 ml to 1.5 ml centrifuge tube, 1 000 r / min centrifugation 10 min, the supernatant as a sample for the test items.

     2.6 Determination of an accurate sample for the test

materials absorb solution 5 μl, injected into high-

    performance liquid chromatography, according to the above chromatographic conditions, determined in accordance with law. Reposted elsewhere in the paper for free download http://

     The linear range of 2.7 to take a concentration of 22.43

    μg / ml and 2243 μg / ml solution of quercetin reference substance, respectively, sample 1,2,5,10 μl, according to the

    above chromatographic conditions were measured to the peak area for the longitudinal coordinates, Quercetin concentration

    (μg / ml) as abscissa, mapping the standard curve, calculated the regression equation: Y = 1.261 458X-16.121 8, r = 0.999 7.

    Showed that quercetin content in range of 22.43 ~ 2 243 ng has good linear relationship.

     2.8 precision test precision drawing 22.43 μg / ml

    reference substance solution 5 μl, repeat sampling five

    times, quercetin peak area value of the relative standard deviation RSD = 0.30%.

     2.9 Stability Test precision drawing 22.43 μg / ml for

    the reference substance and the test sample solution 5 μl,

    respectively, measured at 0,3,6,9 h sample, reference substance and test samples for the peak area of quercetin relative standard deviation RSD were 0.96% and 0.99%. The results show that for the reference substance and test goods

    in the 9 h in good stability.

     2.10 Repeatability test taken six copies of the same sample, according to the above conditions of sample preparation methods and chromatographic determination of quercetin content of the relative standard deviation RSD =


     2.11 recovery tests check samples of known concentration by adding an appropriate reference substance, according to the determination of an average recovery of 101.19%, RSD of 2.35% (n = 6).

     3 Results and Analysis

     3.1 Mount Emei Houttuynia populations of quercetin content in the basic situation of each material by 2.5 Method repeated three times, respectively, for the test materials

precision drawing solution of 5 μl, according to the above

    chromatographic conditions were measured to calculate the

    sample concentration of quercetin in the sample average of the results of determination in Table 3.

     9 populations materials quercetin ranged between 0.24 ~ 2.36 mg / g, average concentration of 1.17 mg / g. Analysis of

    variance results showed that all populations of quercetin content in different materials, F-value of 358.04, more than

    F0.01 = 3.89, up to a very significant level. New Ducan multiple comparison results can be found, in addition to C3 and C4 groups of materials quercetin two bedrooms no

    significant difference, the remaining two bedrooms groups of arbitrary difference between the quercetin have reached a significant level of 1%.

     Table 3 Emei populations at different altitudes Houttuynia quercetin and multiple comparison results (omitted)

     3.2 quercetin content and the relationship between altitude-related analysis showed that the populations of various materials, quercetin and altitude correlation coefficient of 0.215 2, correlation is not significant.

     3.3 quercetin content and the growth of soil pH, and nutrient content of quercetin and its growing relationship between the soil pH, the correlation coefficient between -

    0.437 5, was not significant. And a variety of nutrients and organic matter sub-correlation is also not significant. Its correlation coefficient were as follows: the effective boron (0.607 2), effective zinc (0.394 4), effective manganese

    (0.304 6), total nitrogen (-0.256 6), effective molybdenum

    (0.223 8), organic matter (0.205 2), effective Iron (-0.189

    9), potassium (0.087 3), nitrogen (0.035 8) and the effective copper (-0.034 8).

     4 Discussion

     4.1 Liuwen Yao and other high-performance liquid

    chromatography have been used to buy in Hebeianguo and Beijing had four copies of the material Houttuynia determination of quercetin, its content ranged from 2.0 ~ 4.3 mg / g, an

average of 3.4 mg / g . Chen Li, etc. The project team used

    the same methods from around the country 26 Yuxingcao materials were the determination of quercetin and its content ranged from 0.59 ~ 4.02 mg / g, an average of 1.8 mg / g (to be published). The results of this experiment showed that

    populations of the Emeishan quercetin content of Houttuynia 0.24 ~ 2.36 mg / g, an average of 1.17 mg / g, the overall level of low. The reason may be related to China's extremely rich source of Houttuynia germplasm, genetic background of different, coupled with the growth of different populations related to the ecological environment quite different.

     4.2 Some scholars of plant secondary metabolites and altitude and soil nutrient content, etc. The relationship between ecological factors studied. The results showed that

    the mountain in anisodine henbane and total alkaloid content increases at higher altitude, scopolamine, and atropine Anisodamine were highly correlated with the elevation was not significant; of Aster phenolic flavonoids substances with the

    altitude increases; growth in high altitude, fertile sandy soil of high oil content of fresh valerian [6]; soil trace elements will affect the leaves of Eucommia ulmoides synthesis and accumulation of secondary metabolites. The author Previous

    studies have also found that the Emeishan at different altitudes in the volatile oil from Houttuynia cordata populations bornyl acetate, and limonene content and altitude significant correlation exists between the soil nutrient content of some essential oil components have a significant

    impact (ecological Sinica, to be published). The study found that quercetin content in Houttuynia altitude and the correlation between soil nutrient content are not significant.

     4.3 The tests detected 9 Yuxingcao populations at an

    altitude of 500 ~ 1 600 m, in spite of Houttuynia quercetin and altitude and the correlation between soil nutrient content are not significant. But the experiments also show that, in addition to C3 and C4 groups two bedrooms no significant

    difference in quercetin content, the remaining differences between the various populations have reached a significant level of 1%. Even in the same altitude (1 400 m) mountain valley of two different populations of Houttuynia C1 and D1, its also great differences in quercetin content were 2.36 mg / g and 0.79 mg / g. Speculated that its causes may be related to Emeishan complex terrain, ecological conditions changed

greatly, strongly related to niche differentiation.


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