Background and aims: Medicinal plants are a valuable resource for flavonoids extraction. Prosopis fracta is one of the plants with medicinal properties. Prosopis fracta was found in abundance in southern regions in Iran. Coersetin is medicinal substance found in the fruit of this plant. Quercetin is used in treatment of cancer and viral infections. This study was conducted to determine the Quercetin flavonoid in Prosopis fracta samples in different regions of Khuzestan. Methods: The Prosopis fracta fruit collected from different regions of Khuzestan (Susangerd, Ahvaz, Abadan, Mollasani, Behbahan and Ramhormoz). The beads were isolated from the fruit. Shell and the flesh were dried in an Oven. The dried materials were mixed and flavonoids extracted with a suitable solvent. The extract was injected into the High Performance Liquid Chromatography (HPLC) system. Then, the compound, Quercetin quantity and standard peak in each sample have been determined. Results: Based on the results, the Susangerd samples (0.0033 mg/ml) and Abadan (0.0008 mg/ml) have maximum and minimum levels of Quercetin flavonoid, respectively. Conclusions: Quercetin flavonoid extracted from Prosopis farcta fruits of Susangerd is richer than other regions of Khuzestan province. Therefore, it is recommended to use the Prosopis fracta fruit grown in Susangerd for extraction of the Quercetin flavonoid

Abdali-Mashhadi, Ali Reza.

Delpisheh, Ali.

Direkvand-Moghadam, Ashraf.

Direkvand-Moghadam, Fatemeh.

Ghasemi-Seyed, Vahid.

Lotfi, Amin.

shahrekord university of medical scinces

Advanced Herbal Medicine, 2014; 1 (1): 29-35 *Corresponding Author: Psychosocial Injuries Research Center, Ilam University of Medical Sciences, Ilam, I.R. Iran, Tel: 00988412240404, E-mail: direkvand-a@medilam.ac.ir 29 herbmed.skums.ac.ir                       INTRODUCTION Prosopis farcta is from Leguminosae and sub-family of Mimosoideae, the indigenous of dry and semi-dry areas of America, Asia, and Africa.1, 2 Three species of Prosopis in Iran were founded, including Prosodies cineraria, Prosopis koelziana and Prosopis farcta. Prosopis fracta is a plant of the Leguminosae family and sub-family of Mimosoideae. Prosopis fracta is native to arid and semi-arid tropics of America, Africa and Asia.2 Prosopis fracta is a shrub with  semi-white, hairy, thin and sharp spine branches. Prosopis fracta be found in the provinces of Khuzestan, Gilan, Fars, Hormozgan, Baluchestan, Khorasan and Tehran.3 Prosopis fracta (Fig. 1) has multi medical effects and consider as a main Extraction and measurement of the Quercetin flavonoid of Prosopis farcta in Khouzestan climatic condition  Fatemeh Direkvand-Moghadam1, Vahid Ghasemi-Seyed2, Ali-Reza Abdali-Mashhadi3, Amin Lotfi 3, Ashraf Direkvand-Moghadam4*, Ali Delpisheh4  1Student, Ramin University of Agriculture and Natural Resources, Ahwaz, I.R. Iran; 2Institute of Medicinal Plants, Karaj, I.R. Iran; 3Agronomy Dept., Ramin University of Agriculture and Natural Resources, Ahwaz, I.R. Iran; 4Psychosocial Injuries Research Center, Ilam University of Medical Sciences, Ilam, I.R. Iran. Received: 17/July/2014 Accepted: 11/Nov/2014  ABSTRACT Background and aims: Medicinal plants are a valuable resource for flavonoids extraction. Prosopis fracta is one of the plants with medicinal properties. Prosopis fracta was found in abundance in southern regions in Iran. Coersetin is medicinal substance found in the fruit of this plant. Quercetin is used in treatment of cancer and viral infections. This study was conducted to determine the Quercetin flavonoid in Prosopis fracta samples in different regions of Khuzestan. Methods: The Prosopis fracta fruit collected from different regions of Khuzestan (Susangerd, Ahvaz, Abadan, Mollasani, Behbahan and Ramhormoz). The beads were isolated from the fruit. Shell and the flesh were dried in an Oven. The dried materials were mixed and flavonoids extracted with a suitable solvent. The extract was injected into the High Performance Liquid Chromatography (HPLC) system. Then, the compound, Quercetin quantity and standard peak in each sample have been determined. Results: Based on the results, the Susangerd samples (0.0033 mg/ml) and Abadan (0.0008 mg/ml) have maximum and minimum levels of Quercetin flavonoid, respectively. Conclusions: Quercetin flavonoid extracted from Prosopis farcta fruits of Susangerd is richer than other regions of Khuzestan province. Therefore, it is recommended to use the Prosopis fracta fruit grown in Susangerd for extraction of the Quercetin flavonoid.  Keywords: Gastric ulcers, Medicinal plant, HPLC, Second metabolism. Original article Direkvand-Moghadam F, et al.: Measurement of the Quercetin flavonoid of Prosopis farcta  30 family of medicinal plants.4 Some of medicinal properties of this herb are including the treatment of gastric ulcers, miscarriage, dysentery, rheumatism, laryngitis, dyspnea and chest pain.5 Fleshy parts of the Prosopis fracta fruit have food and drug uses. It is used also for the treatment of bronchitis, asthma, skin lesions, rheumatism, and scorpion sting.3 The active components of Prosopis fracta are included; Lectin, L-arabinose, 5-hydroxyl, Quercetin (Flavonoid), Tryptamine (Alkaloid), Apigenin, Tryptamin and toxin. The combination of Quercetin namely 2-(3, 4-Dihydroxyphenyl)-3, 5, 7-trihydroxy-4H-1- benzopyran-4-one has C15H10O7 molecular composition (Fig 2). Its molecular weight is 302.24. C15H10O7 is dehydrated to form yellow needles from alcohol concentrations at 95 to 97 degrees. One gram of this combination of Quercetin dissolves in 290 ml grain alcohol and 23 ml of boiling alcohol. It is soluble in acetic acid glasyal and yellow in liquid alkaline solution. This compound is insoluble in water and their alcohol test is very bitter.6   Fig 1: Prosopis farcta  Fig 2: C15H10O7 molecular composition  Quercetin has multi medical affects and it is used in the treatment of cancer and viral infections.7 Free radicals in the body cause diseases such as cancer and atherosclerosis and Quercetin clears free radicals. Therefore, Quercetin is beneficial in the prevention of cardiovascular diseases.8, 9  METHODS The Prosopis fracta fruit collected from different regions of Khuzestan including Susangerd, Ahvaz, Abadan, Mollasani, Behbahan and Ramhormoz (Table 1). Details of soil sampling location were shown (Table 2). The beads were isolated from the fruit. The shell and the flesh were dried in an oven at 35°c during 48 hours. The dried materials were mixed and flavonoids extracted with a suitable solvent.  Table 1: Geographical location of the sampling  Location Latitude Longitude Altitude (m) The mean maximum annual temperature (C°) The mean minimum annual temperature (C°) Susangerd 40´    31̊ 6´       48̊ 130 32.5 16.3 Mollasani 36´    31̊ 53´     48̊ 20 36 19.5 Ramhormoz 16´    31̊ 36´     49̊ 150.5 32.7 19.5 Ahvaz 15´    31̊ 33´     48̊ 22.5 32.9 17.6 Behbahan 36´    30̊ 14´     50̊ 313 32.3 17 Abadan 22´    30̊ 15´     48̊ 6.6 32.9 17.9 Advanced Herbal Medicine, 2014; 1 (1): 29-35 31 Table 2: Details of soil sampling location  Location Soil texture EC (ds/m) pH Soil organic carbon (%) Potassium (ppm) Phosphorus (ppm) Susangerd Cilty clay 6.9 7.6 0.86 274 6.8 Mollasani Clay loam 7.1 7.6 0.45 172 6.5 Ramhormoz Loam 6.8 7.8 0.86 182 6.3 Ahvaz Cilty clay 8.7 7.6 0.39 167 6.4 Behbahan Loam 6.8 7.8 0.88 180 6.2 Abadan Cilty clay 10.8 7.8 0.32 207 6.2  The flesh and shell of each sample were grounded separately. Then the powders were used to prepare the test solution. One gram of sample powder was poured into balloons of 250cc. It was added to 50 ml of methanol, 20 ml sterile water and 8 ml 1 normal hydrochloric acid. The sample was placed inside a bath water under reflux and the duration 135 minutes. At this stage, the red color solution was not necessarily representing the reaction. After cooling, the supernatant liquid was poured into 100 ml volumetric flask, and 20 ml methanol was added to the balloons. Then, Reflux was done for 10 minutes. The resulted liquid was passed through the filter paper, and then was transferred into the previous solution 100 ml volumetric flask and the volume was completed with methanol. HPLC is suitable for extracting plant flavonoid Quercetin in Prosopis fracta because of greater sensitivity and better selectivity method HPLC with UV detector. This method most commonly used in comparing to other methods for the analysis of flavonoids. Device specifications of High Performance Liquid Chromatography Extracts were analyzed by HPLC Knauer model. The flow rate was 1 ml per minute. Composition of the mobile phase used methanol, 1% phosphoric acid/ water (40:60). The Filling materials were C8, column length was 25 cm, and the particle size was 5 micrometers. Wavelength NM 270 UV detector was used and the injection volume was 20 ml. The lab temperature was 25 °C and Isocratic forming system was used. Different concentrations 0.01, 0.02, 0.04 (mg/ml) of Quercetin standard in methanol were prepared. Each concentration was injected three times with HPLC, and area under the curve was plotted versus the injected amount. Then, the line equation and correlation coefficient (r2) were calculated from the standard Quercetin. The results reported r2= 1. Then, the Prosopis fracta Shell and the flesh dried extract of the samples of different location were separately using USP method 10, and injected into the HPLC system to identify and specify the amount of Quercetin. The total amount of Quercetin in the samples of different locations was calculated separately. Analysis and validation Different concentrations 0.01, 0.02, 0.04 (mg/ml) were used. Calibration curve analysis is presented (Graph 1).  Direkvand-Moghadam F, et al.: Measurement of the Quercetin flavonoid of Prosopis farcta  32   Graph 1: Calibration curve to the area under the curve concentration of Quercetin  Quercetin chromatogram is presented in (Graph 2). The isolation of Quercetin peak chromatogram of the flavonoids Quercetin in extract samples is presented (Graph 3).   Graph 2: Quercetin chromatogram   Graph 3: Typical chromatogram analysis Prosopis fracta (Quercetin 3.7 minutes)  Advanced Herbal Medicine, 2014; 1 (1): 29-35 33 RESULTSThe Quercetin flavonoid levels were higher in samples with electrical conductivity of the soil, almost neutral or acidic soil and more organic matter. The level of Quercetin flavonoid extracted from Prosopis farcta fruits of the Susangerd, was higher than other regions of Khuzestan province. Therefore, soil quality is likely one of the affecting factors on the Quercetin flavonoid levels in Prosopis farcta fruit. In addition to the climatic conditions of the region, the edaphic condition has an important impact of the changes of the Quercetin flavonoid levels in Prosopis farcta fruit. The Quercetin levels were calculated for each sample and presented in Table 3.  Table 3: Quercetin levels for each sample in different location  Location Quercetin (mg/ml) Susangerd 0.0033 Mollasani 0.0016 Ramhormoz 0.0015 Ahvaz 0.0011 Behbahan 0.0014 Abadan 0.0008  DISCUSSION The Prosopis plant has some of the compounds including: Toxin, Quercetin (Flavonoids), Tryptamine, Apigenin, 5-hydroxytryptamine (Alkaloids), L-arabinose, Lectin.11, 12 Studies showed that compounds like flavonoids have been very valuable for obtaining information about relationships and the hybrid origin of taxa.3-17 Because flavonoids are routinely used for systematic studies, in the present study, we investigated the flavonoid Quercetin levels of different samples obtained from Khuzestan province. The results showed that this concentration is different in samples collected from different locations of Khuzestan province. Based on the results the Susangerd samples (0.0033 mg/ml) and Abadan (0.0008 mg/ml) have maximum and minimum levels of Quercetin flavonoid respectively. As far as, we could not find any Iranian study in the determination of the flavonoid Quercetin levels. A study evaluated the chromatographic characterization of different Prosopis species.18 In another study evaluated the chromatographic variation within the P. juliflora complex,19 and morphology and flavonoid Chromatographies of Prosopis species and hybrids of the Argentinian Chaquena region.20 In other study, Harzallah et al evaluated the patterns of P. farcta flavonoids in branches, leaves, roots, flowers and pods without seeds of two plant populations belonging to this species occurring in the northeast and the southeast of Tunisia. Based the results of this study, a relative similarity was found in the compounds of plants from the two populations.11 Prosopis farcta is widely used by people traditionally for curing lesions in Iran. An Iranian researcher examined the local effect of the fruit dusk powder and root extract of Prosopis farcta on diabetic healing. The results of this study reported the anti-diabetic, anti-inflammatory and healing effects of this plant.21 Other studies have also confirmed the anti-diabetic and anti-inflammatory effects of the Prosopis farcta 22, 23  CONCLUSIONS Quercetin flavonoid extracted from Prosopis farcta fruits of Susangerd is richer than other regions of Khuzestan province. Therefore, it is recommended to use the Prosopis fracta fruit grown in Susangerd for extraction of the Quercetin flavonoid. Direkvand-Moghadam F, et al.: Measurement of the Quercetin flavonoid of Prosopis farcta  34  CONFLICT OF INTEREST The authors declare that they have no conflict of interests.  ACKNOWLEDGEMENT The authors are grateful to thank those researchers who generated useful findings for this study.  REFERENCES 1. Kramer SN. First pharmacopeia in man's recorded history. Am J Pharm Sci Support Public Health. 1954; 126(3): 76-84. 2. Saad B, Azaizeh H, Said O. Tradition and perspectives of arab herbal medicine: a review. 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Extraction and measurement of the Quercetin flavonoid of Prosopis farcta in Khouzestan climatic condition

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