Enayatollah Yazdanpanah*et al. /International Journal of Pharmacy & Technology

Transcription

1 ISSN: 5-66X CODEN: IJPTFI Available Online through Research Article THE EFFECT OF HEIGHT ON EXISTING POLYPHENOLS IN MEDICAL PLANT OF ACHILLEA WILHELMSII C.KOCH Enayatollah Yazdanpanah*, Biology, Payame Noor University, Tehran, Iran. Received on Accepted on Abstract To evaluate the effect of various growing regions on some biochemical and physiological parameters of medical plants, the effect of sea level on poly phenolic compound of medical plant Achillea wilhelmsii C. Koch is studied. The samples are collected from seven regions with different heights in Kohkiliuye and Boyerahmad. The poly phenolic compound is evaluated by HPLC (High performance liquid chromatography). The evaluation of 1 types of poly phenols shows that poly phenols of cinnamic acid, Gallic acid, Caffeic acid, Rutin, coumarins, Vannilin, Carvacrol, Ellagic acid and Eugenol were not found in the samples. Catechin,chlorogenic acid, Quercetin, para-coumaric acid, Trans ferulic acid, Hesperidin acid, Hesperetin, Rosmarinic acid compounds are found. Polyphenols of chlorogenic acid, trans-ferulic acid and Rosmarinic and Catechin had highest level in samples. In the collected specimen from Dena town among the identified compounds, chlorogenic acid was the highest and the highest value is dedicated to Amirabad and the lowest value is Kore. Among the existing poly phenols in Achillea wilhelmsii C.KOCH of Dena town, para-coumaric acid had the highest value in high height and in lower heights, this compound was reduced. In the studied samples of Boyerahmad town, poly phenol of Catechin was reduced with low height. Quercetin had inverse relationship with reduced height. IN the sample of china among the studied regions with the lowest height, Transferulic acid was high and 26.µg ml -1. It is possible that each of studied poly phenols except height is dependent upon other factors as climate, geography, soil, etc. The results of experiments show that different growing regions including sea level can affect cellular metabolism and be effective on phenol compound and medical feature of plant. Keywords: Height, Achillea wilhelmsii C.KOCH, Phenol compound, Metabolism, HPLC IJPT Sep-216 Vol. 8 Issue No Page 165

2 Introduction Today, poly phenols play important role in human health and prevention of different diseases including cancer, cardiovascular diseases and neural diseases and they are taken into consideration of nutrition experts and consumers (Milner, 1; Duthie & Brown, 1). Poly phenols are strong anti-oxidants against oxidative stress of active types of oxygen (Williams et al., 2). Poly phenols or phenolic compounds are distributed widely in plants and 8 phenolic structures are recognized until now, of which more than are dedicated to flavonoids. These natural compounds are found in fruits, vegetable, grain, chocolate and drinks as tea. Most poly phenols are found as glycosides with different sugar units and placement of sugar units in different situations of poly phenol structure in plants (Harborne & Williams, 2; Bravo, 18; Cheynier, 25). Guédo et al. (12) performed a study for the first time on two species of ceretanum A. and A. millefolium and studied for their leaf and flower head flavonoid composition. The main constituents were apigenin and luteolin, mainly found as -O-glucosides and -malonylglucosides (Guédo et al., 13). Lvancheva et al., (22) have studied on species of Achillea ( A. setacea Waldst A. millefolium A.clasiana Waldst A. lingulata A. fraasii LA. clavenae و W.K.A. abrotanoides and major compounds as 6- hydroxyflavonol 3, 6, -trimethyl hydroxyflavones ethers, O-glycosides of luteolin, kaempferol and quercetin were found and C- glycosides of apigenin and luteolin were measured and identified. In this method, the dry plant was mixed with chloroform, then the chloroform was separated and the remainder was dissolved in methanol and was measured by HPLC. By this method, quercetin was identified high (Lvancheva et al., 22). Dadáková et al., conducted a study on A. millefolium and other plants by methanol extraction. In methanol extraction, flavonoid compounds after hydrolysis with Hydrochloric acid were extracted by methanol and then were identified and measured by HPLC. Quercetin is reported as 216±18mg/kg. In another method, providing tea extract, the sample is combined with hot water degree and after being cold is filtered and the liquid under filter is centrifuged and is kept under fridge temperature and is infused to HPLC for measurement and Quercetin was reported as 1.5mg/kDadáková et al., 21). Schieber et al., (25) extracted flavonol of glycosides from mango (Mangifera indica L. Cv. "Tommy Atkins") and identified by high-performance liquid chromatography-electrospray ionization mass spectrometry 6. Among the 22 main analyzed compounds, only kaempferol and quercetin glycosides were traced (Schieber et al., 25). IJPT Sep-216 Vol. 8 Issue No Page 166

3 Bakhshi & Arakawa (Rosengarten, 16) investigated the total phenol and antioxidant activity in in Jonathan apple. There was a positive relationship between total phenol and antioxidant activity (Bakhshi & Arakawa, 26). Materials and Methods 1. Preparation of plant specimen The population of A. wilhelmsii were collected from Dena, Padena, Amirabad, Bahram Beigi, Kare and Boyerahmad, Gelal, Tahloion, Chin in Kohkiloye Boyerahmad. The specimens are washed in the lab and dried in the shade. Table 1- The data of rainfall, temperature and soil of specimen collection sites. Region Dena Boyera hmad Specimen collection site Padena Amirabad Bahrambeigi Kare Gelal Tahlion Chin Sea level (m) Longitude (degree) 51/2 51/ 51/ /82 5/86 5/15 Latitude (degree) 31/2 3/83 3/ /8 31/5 31/1 Soil type Entisols - loam, gravel Entisols - loam, gravel Inceptisols, loam Inceptisols, loam Mollisolsloam Mollisolsloam Mollisolsloam The mean annual rainfall )mm) The mean annual temperat e e( 11/3 1 13/ / The determination of type and amount of poly phenols by HPLC (High-performance liquid chromatography) method. At first, the dried plant is powdered with electric mill and passes fine sieve, then one gram of powdered sample is dissolved in 15mL of methanol. As poly phenols are sensitive to light, the dish of samples is put in aluminum foil to prevent light exposure. The specimens are kept in fridge for 2 h that the specimens are soaked completely and poly phenols exist the powder. Then, the specimens are taken from fridge as covered with foil and they are kept in ultrasonic device that with poly phenol compound waves impact, they exit plant tissue and enter methanol. This is done at temperature for 15min. Then, the specimens exit from the device and foil and after being filtered with filter paper, it is passed from syringe filter. To determine the amount and type of some poly phenols, 2 µl of solution is injected to HPLC. The results are achieved as milligram polyphenol for gram of dry weight by the following Equation: IJPT Sep-216 Vol. 8 Issue No Page 16

4 Mg g -1 DW mg l The features of HPLC in the present study The present study applied HPLC Agilentmodel 12 of Germany equipped to C18, with the length.6 15mm and ID of.5 µm. Methanol and formic acid 1% are used as moving phase. The total time is min, the speed is one millimeter per min, wavelength 28 and 32nm, limit of detection (LOD) is.1ppm and limit of Quamtification (LOQ) is 1ppm. The injection volume is 2microliter and detector is Dynamical absorbance detector (DAD) type. 3-Determination of the amount of some poly phenol compounds in methanol extract of Achillea wilhelmsii C.KOCH by HPLC At first, the standard curve of the following poly phenols are plotted as cinnamic acid, Gallic acid,catechin, Caffeic acid, chlorogenic acidrutin, Quercetin, para-coumaric, coumarins, Carvacrol, Vannilin, trans-ferulic acid, Hesperidin, Ellagic acid Eugenol, Hesperetin and Rosmarinic acid. The Different concentrations of phenol compounds are injected in µg ml -1 extract and the absorption is measured at wavelengths 28, 32nm as shown in Figure s1-1. Then, by absorptions, the standard curve is plotted. It is worth to mention that all above poly phenols have absorption at 28nm except for chlorogenic acid with maximum absorption at wavelength 32nm y = 12.83x R² = Figure 1- Standard curve of Cinnamic acid at wave length 28nm y = 1.65x R² = Figure 2- Standard curve of Gallic acid at wave length 28nm IJPT Sep-216 Vol. 8 Issue No Page 168

5 6 2 y =.211x R² = Figure 3- Standard curve of Catechin at wave length 28nm y = 15.2x R² = Figure - Standard curve of Caffeic acid at wave length 28nm y = 2.112x R² = Figure 5- Standard curve of chlorogenic acidat wave length 32 nm y = 11.81x -.1 R² = Figure 6- Standard curve of Rutinat wave length 28 nm y = 1.2x R² = Figure - Standard curve of Quercetinat wave length 32 nm IJPT Sep-216 Vol. 8 Issue No Page 16

6 area area area area area y = 82.21x R² = Figure 8- Standard curve of para-coumaric acid at wave length 28 nm 6 y = 55.23x R² = Figure - Standard curve of Coumarin at wave length 28 nm y = 1.65x R² = Figure 1- Standard curve of Carvacerol at wave length 28 nm y = 2.x R² = Figure 11- Standard curve of Vanilin at wave length 28 nm 15 1 y = 3.18x R² = Figure 12- Standard curve of Trans formic acid at wave length 28 nm IJPT Sep-216 Vol. 8 Issue No Page 165

7 area area y = 16.8x +.81 R² = Figure 13- Standard curve of Hesperidin at wave length 28 nm y = 1.83x R² = Figure 1- Standard curve of Ellagic acid at wave length 28 nm y = 11.32x R² = Figure 15- Standard curve of Eugenol at wave length 28 nm y = 11.32x R² = Figure 16- Standard curve of Hesperetin at wave length 28 nm y = 2.232x - 11 R² = Figure 1- Standard curve of Rosmarinic acid at wave length 28 nm IJPT Sep-216 Vol. 8 Issue No Page 1651

8 Table 2- The results of evaluation of poly phenols by HPLC at wavelength 28, 32nm in Achillea in different growing regions of Kohkiloye and Buyerahmad province of values in mg in 1 gram dry weight. No. Poly phenol Growing regions name Padena Amirabad Bahrambeigi Kore Gelal Tahlion Chin g ml -1 ( 1g ) )1g 1g ) 1g ) mg/ 1g ) )1g )1g 1 Cinnamic acid 2 Gallic acid 3 Catechin 11/5 12/ /3 2 55/86 / / 3 / 35 11/ 3 /2 3 18/ 65 /61 2/6 / /32 Cafeic acid 5 chlorogenic *acid 83/13 22/ 25 / /1 / / / / / 258/ 8 / / 5 / /3 6 Rutin Quercetin /2 /1 /3 1 5/ 2 18/ 13/ / /3 / / /1 2/6 8 Para- Coumarin acid 3/22 33/5 1 / 1/6 /16 5 2/8 6 / /8 /2 18 / /26 8 3/ 2 /3 1 51/15 Coumarin 1 Carvacerol 11 Vanilin 12 Trans-formic acid / 12/ 1 / / / / 26 /5 112/ / 21/ / 16/ 3 / / 2 13 Hesperidin 66/3 65/ / /6 / /2 /3 2 65/5 8 /2 68 1/ 2 /6 /1 / /8 1 Ellagic acid 15 Eugenol 16 Hesperetin 63/16 2/ 5 / /5 / /3 / /8 /2 62 1/ 3 /16 5 2/ 5 / /2 1 Rosmarinic acid 2/28 2/ / /3 /3 3 6/ 5 / /5 8 / / 2 / /5 / /88 *Absorption is measured at wavelength 32nm..= Not Detected Discussion As shown in Table 2, the evaluation of some of existing poly phenols in Achillea shows that poly phenols of cinnamic acid, Gallic acid, Caffeic acid, Rutin, coumarins, Vannilin, Carvacrol, Ellagic acid and Eugenol were not IJPT Sep-216 Vol. 8 Issue No Page 1652

9 found in the samples. Catechin,chlorogenic acid, Quercetin, para-coumaricacid, Trans ferulic acid, Hesperidin acid, Hesperetin, Rosmarinic acid compounds are found. In the collected specimen from Dena town among the identified compounds, chlorogenic acid was the highest and the highest value is dedicated to Amirabad and the lowest value is Kore. Among the existing poly phenols in Achillea of Dena town, para-coumaric acid had the highest value in high height and in lower heights, this compound was reduced. In the studied samples of Boyerahmad town, poly phenol of Catechin was reduced with low height. Quercetin had inverse relationship with reduced height. In the sample of china among the studied regions with the lowest height, Transferulic acid was high and µg ml -1. It is possible that each of studied poly phenols except height is dependent upon other factors as climate, geography, soil, etc. There are a few studies regarding the environmental effects as growing height in these parameters. In some Cichorium intybus plants as A. millefolium, poly phenol derivatives of caffeoyl as one of the main compounds in aerial type of case is proposed (et al., 211Fraisse). Totally, temperature reduction and increase of UV-B by increasing height can improve biosynthesis of antioxidant phenols as flavonoids in Spermatophytes (Guo et al., 211). Among most of environmental factors, the effect of increase of UV-B radiation and temperature reduction with the increase of height in secondary metabolites of plants are discussed totally. These authors show that sesquiterpene and flavonoids lactones had no significant positive association with growing height. The relative value of flavonoids with free hydroxyl in B flavonoids ring was increased with the height increase. The relationship between total phenol, antioxidant capacity and height changes reported by researchers is not similar. Thus, it seems that in addition to the genetic of cultivars and species, structural difference in phenolic compound and terpenoid and their derivatives were effective on the relationship between secondary products of plant and height (Guo et al., 211; Spitaler et al., 26; Sptaler et al., 28). Acknowledgement: My gratitude goes to all helped me in the study. References 1. Bakhshi D,Arakawa O (26). Effects of UV-B irradiation on phenolic compound accumulation andantioxidant activity in Jonathan apple influenced by bagging, temperature and maturation. Journal of Food, Agriculture and Environment. : Bravo L (18). Polyphenols: Chemistry, Dietary Sources, Metabolism, and Nutritional Significance. Nutrition Reviews. 18:56: Cheynier V (25). Polyphenols in foods are more complex than often thought. American Journal of Clinical IJPT Sep-216 Vol. 8 Issue No Page 1653

10 Nutrition. 81: 223S 22S.. DadákováE, VrchotováN,TřískaJ(21).Content of selected biologically active compoundsin tea infusions of widely used European medicinalplants. Journal of Agrobiology. 2(1): Duthie GG, Brown KM (1). Reducing the risk of cardiovascular disease. In: Goldberg I., editor. Functional Foods: Designer Foods, Pharmafoods, Nutraceuticals. Chapman & Hall; New York, NY, USA: pp FraisseD, Felgiles C, Texier O, Lamaison JL (211). Caffeoyl derivatives: Major antioxidant compounds of some wild herbs of the Asteraceae family. Food and Nutrition Science. 2: Guédo D, Abbe P,Lamaison JL (13). Leafand flower head flavonoids of Achillea millefoliuml. subspecies, Biochemical Systematics andecology. 21(5): Guo XD, Ma YJ, Parry J, Gao JM, Yu LL, Wang M (211). Phenolics content and antioxidant activity of tartary buckwheat from different locations. Molecules. 16: Harborne J.B, Williams C.A (2). Advances in flavonoid research since 12. Phytochemistry; 55: Lvancheva S, Tomas-Barberan F, Tsvetkova R (22). Comparative analysis of flavonoids in AchilleaSp. Sect. Millefolium and Sect. Ptarmica. ComptesRendus de l'academie Bulgare des Sciences. 55: Milner J (1).Reducing the risk of cancer. In: Goldberg I., editor. Functional Foods: Designer Foods, Pharmafoods, Nutraceuticals. Chapman & Hall; New York, NY, USA: pp Rosengarten IRF. (16). the book of spices. Livingston Publication Company Wynne Wood Spitaler R, Schlorhaufer PD, Ellmerer EP, Merfort I, Bortenschlager S, Stuppner H, Zidorn C (26). Altitudinal variation of secondary metabolite profiles in flowering heads of Arnica Montana cv. ARBO. Phytochemistry. 6(): SptalerR, Winkler A, Lins I, Yanar S, Stuppner H, Zidorn C (28). Attitudinal variation of phenolic contents in flowering heads of Arnica montana cv. ABRO: a 3-year comparison.journal of ChemicalEcology. 3: Williams R, Spencer J, Rice-Evans C(2). Flavonoids: Antioxidants or signaling molecules? Free Radical Biology and Medicine. 36: Corresponding Author: Enayatollah Yazdanpanah*, IJPT Sep-216 Vol. 8 Issue No Page 165