Application of Micro-cloud point extraction for spectrophotometric
For Determination of Quercetin in Food samples and aqueous
Abstract
The flavonoids of dietary significance are widely distributed in the plant kingdom and can be categorized as flavones, flavanones, anthocyanidins and isoflavones This study was conducted for quantitative determination of quercetin in Ferulago angulate plant Based on Micro-cloud Point extraction method A fast, simple, inexpensive and effective method is developed for spectrophotometric determination of quercetin based on micro-cloud point extraction (MCPE). The proposed MCPE is essentially a miniaturized form of traditional cloud point extraction (CPE) in which only a few microliters of micellar extracting phase is sufficient for spectrophotometric determination. In this method, to reach the cloud point at room temperature, the MCPE procedure was carried out in brine using Triton X-114 as a non-ionic surfactant. Factors influencing the extraction efficiency, such as concentration of Triton, effect of pH, type of dilution solvent, etc. were investigated and optimized. Under the optimized condition, linear calibration curve for MCPE was in the range of [] and limit of detection of [] was obtained.
Keywords
Ferulago angulate, micro-cloud point extraction, Quercetin, spectrophotometry
1. Introduction
Flavonoids, and particularly quercetin derivatives, have received specific attention as dietary constituents during the last few years. Flavonoids are a large group of polyphenols with a wide range of Pharmacological activities. They are natural products widely distributed in the plant kingdom and generally present in the common human diet. Flavonoids widely distributed in Nature, 4000 flavonoids are found in fruits and flowers. The ant oxidative activity of flavonoids depends on the position and the number of hydroxyl. Evidence has shown that flavonoids are capable of modulating the activity of enzymes and affecting the behavior of many cell systems [1].
Quercetin, or 2-(3, 4-dihydroxyphenyl)-3, 5, 7-trihydroxy-4H-chromen-4-one. It often occurs in them not only in its free form, but too in the form of glycosides. It is found at ppb levels in blood and tissues of animals that received plant feeds [2]. Quercetin can be produced by hydrolyzing rutin in hydrochloride aqueous solution [3] .Amidst 180 various glycosides of quercetin, found in nature, the most prevalent is rutin, that is, quercetin.[4-5] Quercetin, 3, 30, 40, 5, 7-pentahydroxylflavone, is one of the most abundant flavonoids present in vegetables and fruits . Quercetin (Figure 1) also can protect human DNA from oxidative attack in vitro. As all flavonoids, quercetin show heavy antioxidant activity, due to free radical scavenging [6] .antioxidant effect quercetin implied to be helpful for human health. The epidemiological studies have shown that possible role quercetin exhibits anti-cancer[7-9] , anti-inflammatory, stabilizes cell membranes, antihistamine, anti-allergic, anti_edematous, antiviral activity, and may also prevent cardiovascular illness [11] in humans.Because quercetin have beneficial activities, the determination of QR has been interesting research area in last decade. Several methods have been developed for the determination of LODs by HPLC with fluorescent [12-13] and MS [14] detection, capillary electrophoresis [15] HPLC coupled with chemiluminescence (CL) [16] spectrophotometry [17] and electrochemical methods [18]. Lunate et al. [19-20] have identified flavonoids in beverages by liquid chromatography with electrochemical detection (LC-ED). High-performance capillary electrophoresis (HPCE) was used for determination of the pharmacologically active flavonoids in Ginkgo balboa and phytopharmaceuticals. These methods are highly sensitive and effective; yet, they also have disadvantages, as complexity of operation, reagent-consuming, and high cost.
Apart from flavonoids’ therapeutic effects, quercetin is also excellent an scavenger of free radicals [6] .To overcome these problems, we decided to operate the spectrophotometric determination by means of micro quartz cell. Following our researches, here we applied MCPE as a very fast, simple, inexpensive and environmentally friendly method was used for preconcentration and determination Quercetin In this work we have studied the influence of quercetin and a number of other compounds. For this purpose, from using Triton X-114 as a non-ionic surfactant. Onion, tomato, Orange juice, Apple juice chosen as the examples of Food Samples.
Figure 1. Structure of quercetin
2. Experimental
2.1. Instrument
A digital pH-meter (HANNA instruments HI 2211 pH/ORP Meter) equipped with a glass-combination electrode was used for the pH adjustment. A UV-Visible (Spectrophotometer UV–Vis 2501 PC, Shimadzu, Kyoto, Japan) for measuring the absorbance and recording spectra. The remaining particles in the tube isolated by centrifugation (Hectic ZENTRIFUGEN ROTOFIX 32 A).
2.2 Materials and reagents
All chemicals and reagents used for experiments and analyses (Deionized water, Triton X-114, Na2SO4, Quercetin, Methanol (were of analytical grades and purchased from Merck (Darmstadt, Germany) or Aldrich (Chemical Co., Milwaukee, WI, USA). Triton X-114 (5% v/v) and Na2SO4 (5% w/v) solutions were prepared in Deionized water.
2.3. Micro cloud point extraction procedure
Immediately after addition of salt, the solution became cloudy. It left to stand in a water bath at 40°C for 10 min.The obtained cloudy solution was centrifuged for 5 min at 5000 rpm. During the centrifugation the surfactant settled down at the bottom of the test tube to form an enriched surfactant phase. The separated phase was diluted with 0.5 mL methanol at the bottom of the tube. Then this mixture was transferred to a microcell for spectrophotometric determination
2.4. Extraction and hydrolysis of Food samples and aqueous
a. Samples prepared from onions and tomatoes were washed and dried for 3 hours at 110 ° C o. Each dried material weighed 0.5 g and soaked in methanol for 24 hours. Then the specimens were placed in ultrasonic. To obtain a smooth solution of the specimens in conical tubes centrifuged for 5 min at 5000 rpm, after separating the supernatant solution and evaporating the solvent, the residue was covered with methanol and distilled off in a dipped water dipped 50ml.
b. Apple juice from the local market were diluted 10-fold with water. Centrifuged for 5 min at 5000 rpm to obtain a nearly colorless solution. Orange juice from the local market was
Centrifuged for 5 min at 5000 rpm.
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