INTRODUCTION:

One of the most popular techniques nowadays is Pressurized Liquid Extraction (PLE), also known As Accelerated Solvent Extraction (ASE). The technique is today well-established and has been used for the extraction of a great variety of compounds from numerous matrices.

Received on 28.04.2015 Modified on 06.05.2015

Research J. Pharm. and Tech. 8(6): June, 2015; Page 800-810

DOI: 10.5958/0974-360X.2015.00129.8

Compared to classic Extraction in a Soxhlet apparatus, complete PLE can be achieved in shorter time with a small volume of organic solvent and much better penetration of sample by the solvent. Extraction parameters are Thoroughly investigated in order to produce an exhaustive methodology, but exhaustiveness very often Leads to co-extraction of unwanted, interfering matrix components, thus the selection of the proper Solvent which must be able to solubilize the analyst and minimize co-extraction of other matrix Components, is very important. (PLE) is a new sample extraction method that employs liquid solvents at elevated temperatures and pressures to prepare samples for analysis by either gas chromatography or liquid chromatography. Pressurized liquid extraction is similar to Soxhlet extraction, except that during the extraction process the solvent condition inside the PLE cell approaches the supercritical region which results in more efficient extractions. The elevated temperature allows the sample to become more soluble and achieve a higher diffusion rate while the elevated pressure keeps the solvent below its boiling point. At elevated pressures and temperatures solvents can penetrate solid samples more efficiently which reduces solvent usage. A pressurized liquid extraction when compared to a traditional Soxhlet extraction shows a reduction in extraction time to 22 minutes from 18 hours and a decrease of total organic solvent consumption to 80 mL or less of organic solvent from 300 ml.

Fish muscle tissue containing 3.2% (w/w) lipids and native polychlorinated biphenyl organ chlorine pesticides and other related compounds was used for testing. Purification of crude extracts was carried out by gel permeation chromatography employing Bio-BeadsS-X3. Identification and quantitation of target indicator PCBs and OCPs was performed by high-resolution gas chromatography with two parallel electron-capture detectors (ECDs). Classical methods for the determination of trace pollutants in environmental solid samples are usually laborious and time-consuming multi-step procedures

Fig.1:Instrument of PLE

They usually require many manual sample handling steps. At line or on-line coupling of these steps is one of the Main goals of modern analytical chemistry. Several examples of on-line clean-up procedures can be found in the literature. However, the analyst extraction itself is usually regarded as the most difficult step when developing completely on-line and/or automated procedures for solid or semi-solid environmental samples. Because of the low levels at which micro contaminants are generally present in the Environment and the variety of the samples, the selected extraction technique should be essentially exhaustive and, preferably, easy to standardize. Preference for techniques such as Soxhlet or Soxtec extraction rather than more selective, But also highly analyze- and/or matrix-dependent, techniques such as supercritical fluid Extraction. Microwave-assisted solvent extraction (MASE) and pressurized liquid extraction are generally faster, and less analyte and matrix-dependent and provide cleaner extracts than conventional methods involving heat treatment. These characteristics have caused both techniques, and specifically PLE, to be frequently used as extraction procedures for a variety of Environmental applications. However, they are always carried out off-line. The at-line, or online, coupling of MASE or PLE with the separation-plus-detection part of the system would require miniaturization of the extraction devices and, if at all possible, no additional clean-up Step. Regarding the latter aspect, PLE has the advantage over MASE that no additional Filtration step is required. The method for the determination of 2 widely administered anticancer drugs, ifosfamide and cyclophosphamide, contained in sewage sludge. The method relies on the extraction from the solid matrix by pressurized liquid extraction, sample purification by solid-phase extraction and analysis by ultra-high performance liquid chromatography coupled with tandem mass Spectrometry. The different parameters affecting the extraction efficiency were optimized using an Experimental design. Solvent nature was the most decisive factor for the extraction but interactions Between some parameters also appeared very influent. The method was applied to seven different types of sludge for validation.

The performances of the analytical method displayed high variability between sludge’s with limits of detection spanning more than one order of magnitude and confirming the relevance of multi-sample validation. Matrix effect has been determined as the most limiting analytical step for quantification with different extent depending on analyte and sludge nature. For each analyte, the use of deuterated standard spiked at the very beginning ensured the complete compensation of losses Regardless of the sample nature. The suitability of the method between freshly spiked and aged samples has also been verified. The optimized method was applied to different sludge samples to determine The environmental levels of anticancer drugs. PLE method was developed by using conventional high performance liquid Chromatography instrument and the performance of the PLE system was tested using environmental soil Samples. Since PLE was demonstrated to be very efficient, the technique was rapidly accepted by the US Environmental Protection Agency (EPA) as a method for evaluation of solid wastes. Often the organic Solvent or combination of solvents utilized in existing Soxhlet method can simply be adopted by the PLE Method. Consequently, the year after the first publication dealing with persistent organic pollutants (POPs) in Soils and sediments were presented, the number of publications dealing with PLE of POPs has increased, where the main focused on PAHs Pressurized liquid extraction (PLE) is a newer technique which has been successfully applied to extract a variety of organic compounds from complex solid samples. Traditional solvent techniques use large quantities of toxic organic solvents, are labor-intense, need long extraction times, possess low selectivity, and/or low extraction yields and can expose the extracts to excessive heat, light, and oxygen. Opposite to that, PLE uses less solvent in a shorter period of time, is automated, and involves retaining the sample in an oxygen and light free environmental conditions. Such techniques, like supercritical fluid extraction (SFE), are frequently used to obtain functional compounds from natural sources, however, PLE has not been widely applied as a routine tool in natural product extraction but recent studies have demonstrated the advantage of PLE for the extraction of natural compounds from different matrices(fig 1). PLE provides higher selectivity, shorter extraction times and frequently does not require large amounts of toxic organic solvents; furthermore, water can be used as medium polarity solvent when working at high temperatures and moderate pressures, that is, at subcritical conditions. A drawback of PLE is its requirement of especial instrumentation in order to get relative high pressures together with high temperatures. Moreover, there are not data available on solubility of natural compounds in the solvent at the pressures and temperatures employed in PLE.

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Fig 2: High speed pressurized liquid extractor

The PLE™ system is a high-speed pressurized liquid extraction system designed to perform sample extraction of multiple samples simultaneously (fig 2). The PLE system delivers high recoveries and excellent precision for all analyses in minutes instead of hours. Inexpensive stainless steel extraction cells with end cap filtration keep operational costs at a minimum. Optional disposable end cap filtration increases productivity and saves valuable time. Pressurized fluid extraction is similar to Soxhlet extraction, except that the solvents are used near their supercritical region where they have high extraction properties. In that physical region the high temperature enables high solubility and high diffusion rate of lipid solutes in the solvent, while the high pressure, in keeping the solvent below its boiling point, enables a high penetration of the solvent in the sample. Thus, PFE permits a high extraction efficiency with a low solvent volume (15-40 ml) and a short extraction time (15-20 min). That procedure is also known as "Accelerated Solvent Extraction" (ASE). This procedure was first developed by Dionex¹and validated on a commercially-available, automated extraction system (Dionex ASE).

With the same solvent mixture employed in the Folch procedure, the pressurized solvent extraction of total lipids in poultry meat reduced the consumption of solvents and the time extraction while it has given similar lipid recoveries and fatty acid compositions²The PFE device includes an extraction cell (1 up to 100 ml) maintained at a temperature between 80 and 200°C into which a solvent is pumped and maintained at 10-20 MPa for some minutes. Then, the extract is pushed into a collection vial by a second volume of solvent and finally the whole solvent is pushed with an inert gas flow. As for SFE, an inert matrix must be added if some moisture is present in the sample. Lower pressure and solvent temperature are used in the ANKOM fat analyzers. A batch processing can be accomplished by encapsulating the samples in filter bags. The filter bag and sample are weighed before and after the extraction, the weight loss corresponds to the lipid content PFE was initially used for environmental contaminants (EPA Method 3454: herbicides, pesticides, hydrocarbons) in soils, sediments and animal tissues but is now frequently used for food (meat, seeds, feeds), pharmaceutical products, and several other biological samples.

This technique was shown to be suitable to replace the Folch extraction oxysterols in food³and carotenoids in plants⁴.The efficiency of extractions with pressurized solvents (hexane, methylene chloride, isopropanol, and ethanol) of polar and nonpolar lipids was examined in corn and oats kernels⁵.The effects of solvent polarity and temperature were tested on the recovery of total lipids, triglycerides, glycolipids, and phytosterols.PFE was used to isolate tocopherols from several seeds and nuts and was shown to yield very clean extracts and recoveries similar to conventional techniques⁶Optimized extraction parameters were described for the maximal extraction efficiency of cereal tocopherols and tocotrienols⁷ or carotenoids from a migroalga⁸.A new fractionation scheme for lipid classes was proposed⁹Basically, a two-step pressurized liquid extraction was combined with an in-cell fractionation using silica-based sorbents (silicic acid or cyanopropyl silica) placed at the outlet of the separation cartridge. The extraction consists of 2 cycles of hexane/acetone at 50°C to obtain neutral lipids followed by 2 cycles of chloroform/methanol at 110°C to obtain polar lipids. Using a homemade pressurized liquid extraction system, good efficiency in the determination of carotene, tocopherols and tocotrienols in residue oil from palm fiber was also reported ¹⁰. The relative extraction recoveries (~100%) were found to be equivalent to conventional Soxhletextraction.PFE has not been widely applied as a routine tool in natural product extraction but as it uses less solvent in a shorter period of time and in oxygen and light-free environment, it has the potential to be a powerful tool in industry. An adaptation of classical extraction procedures for use with PFE toward the use of about 100 mg of tissue was proposed¹¹using gravimetry and fatty acid analysis, it was proven that efficient results were obtained with chloroform/methanol (60/40) as solvent mixture for fish tissue extraction (fig 3).

Fig 3:PLE extractor assembly and components

A comparison between PFE and the Folch procedure for the extraction of cereal, egg yolk and chicken muscle revealed that higher fatty acid contents were obtained with a mixture of isopropanol/hexane for cereal and egg yolk, however, a chloroform/methanol mixture was more efficient with muscle samples¹². A comparison with soxhlet and ultrasound-assisted extractions showed that pressurized liquid extraction was more effective for terpenes, fatty acids and vitamin E contained in leaves of Piper sp¹³. The quantitative measurement of fatty acids in human plasma and erythrocytes has been efficiently made using solvent extraction in an ASE system¹⁴. Pressurized extraction of sterols in marine sediment samples was shown to give recoveries higher in comparison to soxhlet extraction¹⁵. Detailed applications notes on the extraction of fats from dairy products, meat, chocolate, oilseeds, and various food matrices can be downloaded from the Dione web site. As the operating conditions of PFE are similar to those of Soxhlet extraction, this technique is continuously in development despite the need of an expensive specialized equipment

PLE of Ginger with Bio-Ethanol:

Ginger plays an important role in the Indian spices and it is widely used as traditional and contemporary medicines ¹⁶. It is extensible used in the formation of various product like pickle, chutney, ginger beer, ginger wine, biscuit etc¹⁶.It has various biological action such as anticancer, ant tissue, cardio tonic, antifungal etc.^17,18. Extraction were done by using solvent under the specified pressure i.e. 1500 psi for 5min in a 2 cycle. First cycle is known as static cycle it takes about 3-5 min for loading the sample and the second cycle takes about three min for solvent purging and cooling, depressurizing and unloading takes 6-7 min therefore the full process take 20 min for extraction. In which ginger sample is mixed with solvent under 100°c and the final yield is obtained which is more than any other method. Generally Bioactive compounds are gained from steam distillation and solvent distillation which is time consuming, laborious, required large number of solvent and to our come this is new technique or method is used i.e. PLE (pressurized liquid extraction) which is having many advantages i.e. it requires small amount of solvent, fertilizer extraction, less time consumption^19,20.

Pressurized Liquid Extraction as a Green Approach in food and Herbal Plants Extraction:

Now-a-days herbal product and their exaction became more popular. PLE is the “green technology” use for the extraction. And the author has discussed the extraction principle and their parameters which can improve the process²¹. Traditionally, for the extraction of the bioactive constituents includes Soxhlet extraction, sonication, blending and solid-liquid extraction which have several drawbacks like required less amount of sample, it apparatus is costly and it cause human health issue^22,23But the green technology PLE extraction has cover all the extraction process. It has several advantages over all the process due to popularity and intense in the extraction of bioactive compound from herbs. It has more advantages over traditional method, PLE enhance the extraction field and also decrease the time and consumption of solvent. This technology has one drawback that it is expansive but this drawback overlooked due to importance of antioxidant as diet^24,22. In this method of extraction, extraction is carried out in an elevated temperature and pressure with liquid solvent and extraction takes place in a room temperature^23,25and when hot water is used in the experiment is known as pressurized water extraction (PHWE).

Application of Derivatization Reaction to Trace organic Compounds during Sample Preparation based on Pressurized Liquid Extraction:

PLE is the versatile method for the extraction of analysts and modern approaches combine for PLE with in-situ derivatization, food, environment and biological analysis. Traditional extraction technique requires more extraction time, large amount of sample, causes human health issue. Another drawback of this that is required concentrated extraction. More our, the extraction of compound from the solid sample are sensitive, thermos labile and obtain in low concentration which is not convenient for the extraction process.²⁶But PLE extraction process have various advantages i.e. faster extraction with less consumption of solvent, required less time, handling easy. And with the combination of derivatization involves various advantages such as elevated temperature and parameter permit for efficient extraction. It also increase the rate of reaction and decreases the time of reaction so, PLE has more useful and better advantages process then the other traditional extraction process.

Enrichment of Antioxidant Compounds from Lemon balm by Pressurized liquid Extraction and Enzyme assisted Extraction:

Lemon balm is traditionally grown in the central and Mediterranean Europe²⁷.And it grown and collected throughout the world due to their wlinary properties. Lemon balm possess various pharm logical activities like antimicrobial activity²⁸, sedative ²⁹ antioxidant activity³⁰, and many more studies have been done on lemon balm and have shown phenolic compound, tennis are most advantage present in the plant³¹.

PLE is very easy technique for regaining of bioactive compound. It has many extraction process but for more efficient and to improve the quantity of extraction enzymes are mixed which enhance the yield reduce the time and also increase the antioxidant capacity and which are very helpful to identify different derivatives present in the plant. The PLE is takes place in an extractor which is having solvent controller unit. At start of the day of experiment solvent is solicited for 10 min and the solvent i.e. used in this one ultra-water and ethanol kept for extraction is completed it is collected and packed in a stainless steel container and mixed with the 2 gm sea.

Optimization of Pressurized liquid Extraction of Inositol’s from Pine Nuts:

Now a days PIE being used for the extraction of various things like persistent organic compounds from food and environment. It scarcely applicable in literature but unlike other, PLE introduced a bioactive extract inositol’s from pine nuts which are rich in liquid and antioxidants. Amounts of chiro-inositol, pinitol and several glycol cydilitols, galactopinitol, fagopyritol B1, and other glycosylinositol are also founded in it³². Inositol’s is a kind of carbohydrates present highly in all vegetables but low in meats it has many bioactive properties like helps in grow thing of microorganisms act as lipotropic agent for animal and part of phosphatidylinositol. Separation procedure is done conventionally through strong anionic exchange resins of hydroxide form and simulated mixing breed chromatographic system for special cases. The most effective production of inositol extract; free from interfering sugars from legumes. Experiments takes place in an oven with a temperature another which is made up of stainless steel. It also consist of an isocratic pump which provide the required pressure and another value fitted in outlet of the extraction cell. Cell was scaled. Now the pine nuts is crushed in the mortar and mixed with the sea sand in 1:2 proportion and set for the extraction^33,34. It is being selected the most appropriate solvent because it needs only 18 minute and 3 cycles of 1.5 nil water each one in 50°c and 10mpa which is optimum. At present inositol plays a vital role as insulin for treatment of diabetes, mellitus, and obesity atherosclerosis from business point of view^35,36also it is the optimum production as requires automatic equipment which is less manipulation and straight forward to industrial scale. Hence inositol considerably a best alternative methodology with optimum resource and less time consuming.

Determination of three Curcuminoids Curcuma wenyugin Y.H Chen efc. ling by Liquid Chromatography with PLE:

Curcuminoids is a type of human diseases with its regular pharmacological effect. It is famous for its low toxicity character which helps in curing diseases like hepatitis, menstrual disorders. It is also observed that, various pharmalogical activity such as anti-inflammation, anti-microbial, anti-oxidation act present in it due to quality control of raw herbal material and medical products but all the same time ³⁷.It also possess various pharmalogical activity such as ant inflammation, antimicrobial, antioxidant ion activity present³⁸due to quantity of raw herbal material and medical products but at same time. It is also obtained as an extract with developed simultaneous, identification of three curcuminoids in curcuma wenyugin. Several different types of separation method were applied for the extraction of cur cumin such as HPLC³⁹, capillary electrophoresis ⁴⁰TLC among all this extraction method PLE has become the most famous technique. PLE was performed using methanol, temperature set 100° c, pressure at 1500 psi. Comparatively it takes lesser time and solvent consumption. In these one pressuring, two daughter ions are used.Greater extraction efficiency was obtained by an optimized PLE procedure. More over method provide useful procedure experience and shows promising perspective in analysis of other sample.

Determination of Saponins and Fatty acid in Ziziphus jujuba:

In china Suanzaoren is one of the most commonly used Chinese medicines which has been used for its effect on insomnia and anxiety⁴¹Suanzaoren have several biological activity such as hypnotic sedative, hypotensive, anti-hypoxia, anti-hyperlipidemia and hypothermic showed by the modern pharmalogical studies⁴². Generally, the traditional Chinese medicine have the different therapeutic effect which gives an integrative result of various biological compound. Both sapiens and fatty oil are responsible for its therapentic activities in suanzaore fatty acids are present in large amount which is considered as a major active fraction for sedative and hypnotic action^43,44.To control the quality of Sunanzaoren, sapiens and fatty acid plays an important role and for the determination of sapiens, HPLC with VV detection is the prevailing technique. However, due to the low wavelength vv is used for detection of saponins⁴⁵.In addition linoleic acid have different types of pharmalogical activities, sometimes in it used for the treatment of cancer⁴⁶. It is also play important role in the treatment of increase endurance exercise capacity and positively benefitting bone mineral density in women.The evaporative light scattering detector does not depend on the sample which eliminates the common problem associated with other HPLC detector and in the current study, saponins and fatty acid in suanzaoren and saponins by using simple method HPLC-ELSD and PLE.

Hence, the PLE procedure improve on and different types of parameters are include i.e. the type of solvent [methanol, ethyl acetate methanol] with interval of S. particle size (20-40 mesh, 40-60 mesh and 60-80 mesh). Pressure, temperature, time, flush time and cycles of extraction. Fatty acids are determined by GC-MS as the esters after derivatrization. PLE procedure was determine by performing PLE on the same sample for the optimized condition which is calculated based on the total of individual investigated compounds.PLE were performed on the system and the powder of jiaughuang or yujin was mixed in an equal proportion in a stainless steel cell with solvent (methanol) in a 140°c temperature and 1000 psi pressure for 5 min. after extraction it is filtered and finally product is obtained.

Determination of Eight Component Rhizome and Tuberous root of Curcuma longa using PLE:

In South-Asia curcuma longer in inherent plant which is cultivated largely throughout the warmer parts of the world. Now a days it is cultivated large scale in India and China. It is used in powered form and to enhance the flower and color of the food it is widely used as a food additives⁴⁷C.longa are also widely used as two Chinese medicine e.g, Jianghaung and Yujin. C. longa is also have various biological activity i.e. anti-oxidation^48,49antimicrobial^50,51, anti-atherosclerotic^52,53, antinflammatony^54,55. They have different biological activity and chemical component⁵⁶. Due to the absence of chemical standard, the components of C. longa cannot be determine but through the LC-MS GC and GC-MS components of turmeric oil have been determined⁵⁷or the estimation of eight different components such as β-caryophyllene, AR-turmerone, β-sesquiphell and rexendrene, AR-turmerone, α-turmerone and β-turmerone the method PLE and GC-MS was provided. Hence, the conclusion is the determination of essential volatile compounds in rhizome was preferred due to their simple, rapid and accuracy which is helpful to control the quality by the 4 components AR-WR cumene, AR-turmerone, α-turmerone and β-turmerone of rhizome and tubenose tube.

As an Alternative Process to Antioxidant Carotenoids Extraction from Haemato coccuspurialis:

Carotenoids is extracted from the Haemato coccuspluuialis which is green motile unicellular algae⁵⁸and in the present year it is widely popular as nature source of the carotenoid. It also possess many different pharmalogical activities like anti-inflammatory, in the treatment of cancer, in anti-aging cosmetics and it is now-a-days used in the food colorant in agriculture and poultry. Its main component which it consist of is anti-oxidant capacity⁵⁹.And for the extraction many different extraction method applied by using mainly organic solvent^60,61. But all the methods are very difficult to apply because it need more amount of sample⁶². Over this PLE is introduced as a green extraction technique for the extraction of bioactive compounds. PLE is used due to many advantages over all other traditional methods because it use less amount of solvent, it controls the temperature and pressure and also it required time for extraction as compared to other process. The process are performed in four different temperature (50°, 100°, 150°, 200°) for 20 minutes. Ethanol plays an important role for getting best yield with the best anti-oxidant activity but in low temperature and hence PLE is better method for extraction and it is widely performed in the industry in recent years because of its fast and clean extract. Extraction is occurs in the solvent extractor with two different solvent ethanol and hexane in a different temperature of 20 min and there whole process repeat three times.

PLE of Vitamin E from Grape Seed oil:

Vitamins are biochemical which are obtained from the different plants and animal vital nutrients than an organism requires in limited amount. Among all the vitamin A, vitamin E is present in eight different form which is fat soluble ^63,64and it is widely obtained from grape seed oil from wine industry it’s having high anti-oxidant activity which present the rancidity of oil while storage^65,66, and also used in the industries as food additives and cosmetic⁶⁷. Vitamin E is the main digestible source. It also contain to copherls and tocontrienols in very small amount. Buttocotrienols the most active form.PLE is the method which gives the fast and efficient extraction of analystes by comparing temperature and pressure which compiles the temperature and pressure⁶⁸2005 and also used for the extraction of many vegetable oil lipid and fatty acid^69,70because of its short time requirement. In this the process is performed for 30 minute at 1500 psi pressure and 100°c temperature and the yield kept in glacial 250ml and collected in collector and again send for the purification or after concentrated by nitrogen. Therefore PLE gives the efficient result for the extraction of oil because it is very simple fast and vegetable lesser time and consumption simple it is more preferable.

Other Conventional Method of Extraction:

• Soxhlet extraction is a piece of laboratory apparatus invented in 1879 by Franz van soxhlet. It was originally designed for the extraction of a lipid form a solid material⁷¹.

• These experiment so uses a soxhlet method to determine the liquid contents of plants and animal tissues.

• The Soxhlet method for determining crude fat content is a lengthy process requiring up to a day for a single analysis. The solvent extraction step alone takes 6 hours (fig 4).

• The method is therefore not favored for routine testing process in the meat industry, rather it is used as a standard reference⁷¹

Fig. 4:Soxhlet Apparatus

1. Stirrer bar/anti-bumping granules, 2.Still pot-still pot should not be over filled and the volume of solvent in the still pot should be 3-4 times the volume of the Soxhlet chamber, 3.Distillation path, 4. Soxhlet thimble, 5.Extraction solid, 6. Syphon arm inlet, 7. Syphon arm outlet, 8. Expansion adapter, 9.Condenser, 10.Cooling water in, 11.Cooling water out.

Pressurized Liquid Extraction as an Alternative to the Soxhlet Extraction:

The aim of the present study was to develop a pressurized liquid extraction (PLE) method as an alternative to the relatively time consuming Soxhlet extraction procedure described in the United States Environmental Protection Agency (US EPA) method TO-13A for the extraction of PAHs adsorbed onto polyurethane foam plugs (PUFs). For this purpose PUF air samples were collected and split into two parts: one part extracted using PLE and the other one using Soxhlet extraction. Comparable PAH concentrations were obtained upon analysis of the extracts showing that the PLE method developed in this work is a more convenient choice than the commonly used Soxhlet extraction technique proposed by US EPA for the determination of PAHs in air samples. In fact, the developed PLE method required shorter assay times (minutes versus hours), less solvent consumption and simpler operational methods. The exhaustiveness of the developed PLE method was evaluated using repeat static extraction cycles, demonstrating an extraction efficiency for the PAHs of greater than 99%. The PLE method was then applied to diesel exhaust and wood smoke PUF samples showing an extraction efficiency for the PAHs of greater than 93% and 96%, respectively. Furthermore, a PLE method for PUF cleaning was developed as well and employed as an alternative to Soxhlet extraction. The PLE methods developed for cleaning and extracting PUFs presented in this work are suitable to be used in mutagenicity studies using the Ames Salmonella assay as no mutagenicity was found in the PLE generated blanks⁷²

Principle of PLE:

To understand any extraction technique it is first necessary to discuss. Some underlying principles that govern all extraction procedures. The chemical properties of the analyst are important to an extraction, as are the properties of the liquid medium in which it is dissolved and the gaseous, liquid, supercritical fluid, or solid extract ant used to exact a separation. Of all the relevant solute properties, five chemical properties are fundamental to understanding extraction theory: vapor pressure, solubility, molecular weight, hydrophobicity, and acid dissociation. These essential properties determine the transport of chemicals in the human body, the transport of chemicals in the air–water–soil environmental compartments, and the transport between immiscible phases during analytical extraction. Response surface methodology (RSM) was used for the determination of optimum extraction parameters to reach maximum lipid extraction yield with yeast. Total lipids were extracted from oleaginous yeast (Rhodotorulaglutinis) using pressurized liquid extraction (PLE). The purpose of this study was to develop an analytical method for the determination of organ

ADVANTAGES:

· Pressurized liquid extraction (PLE) improves the extraction yield, decreases time and solvent consumption, and protects sensitive compounds.

· PLE facilitates the use of solvent mixtures and other extraction additives that would enhance the extraction efficiency.

· The use of combined and hyphenated results in a substantial decrease in the amount of sample used with better efficiency and selectivity.

· Different studies discuss using PLE to enrich phenolic compounds, lignin’s, carotenoids, essential oils and other nutraceuticals from foods and herbal plants Pressurized liquid extraction (PLE) is a relatively new technique applicable for the extraction of persistent organic pollutants from various ma-Trices.

· PLE is fast extraction process than any other traditional method.

· It takes lesser time.

· It gives better yield then other extraction process.

· It provides improved quality of extraction and it is handled very easily.

The main advantages of this method are short time and low consumption of extraction solvent. The effects of various operational parameters (i.e. temperature of extraction, number of static cycles and extraction solvent mixtures) on the PLE efficiency were investigated in this study.

Recent Advancement in Pressurized Fluid Extraction:

Pressurized fluid extraction (PFE) is a technique performed to extract solid or semi-solid samples using organic solvents. Elevated temperatures (up to 200 °C) are used to increase the kinetics of the extraction process while applying high pressures (for example, 1500 psi) to maintain the organic solvents in the liquid state. PFE is unique in that extractions are performed rapidly with reduced solvent use, compared with traditional extraction techniques. For example, PFE can reduce the extraction time down to 20 min per sample versus hours using Soxhlet and reduce solvent consumption to 30 mL per sample.

PFE instrumentation follows a common pathway to produce extracts. An extraction cell containing the sample is loaded into an oven and a pump transfers extracting solvent into the cell from a reservoir. The cell is then pressurized and heated to a preset temperature. The temperature and pressure in the extraction cells rises above ambient levels and the hot solvent enhances the extraction rate of the analytes from the matrix. PFE systems are designed so that solvent will flow through the extraction cell and be collected into a bottle or tube at the end of the flow path. Once the extraction is complete, the extraction cell is purged with nitrogen gas to remove residual solvent and the collected extract is ready for concentration and analysis. PFE systems are currently manufactured by three vendors:

Thermo Scientific (the accelerated solvent extraction [ASE] system); Fluid Management Systems, Inc. (FMS) (the pressurized liquid extraction [PLE] system); and Büchi (the Speed Extractor system). Each system offers automation capabilities for the analytical laboratory to reduce the amount of time spent on sample preparation. These systems use elevated temperature and pressure to improve extraction efficiency and productivity compared with traditional extraction techniques such as Soxhlet. A summary of each system along with significant features is provided in this article. Pressurized liquid extraction (PLE) is widely used in persistent organic pollutants (POPs) extraction from environmental matrices such as soil and from biological matrices such as tissue. The FMS PLE instrument makes use of elevated temperature and pressure to increase the rate and efficiency of the extraction process, producing extracts in as little as 20 min. Up to six samples can be extracted simultaneously using the system's parallel extraction mechanism. The system is modular, can be expanded from one to six channels, and uses stainless steel extraction cells (5–250 mL) with end caps that contain Teflon filters. DMS-6000 Editor software is used to control the instrument, store multiple extraction methods, and plot temperature and pressure data for each channel used. The PLE instrument can be interfaced with both in-line clean columns and an evaporator that combine the extraction, cleanup, and concentration procedures into one step. In-line cleanup of extracts is accomplished with the use of in-cell packing materials (for example, silica or carbon), and by interfacing with columns that are packed with adsorbents (carbon, silica, and alumina). A flow-through design is used to move solvent downward through a heated and pressurized extraction cell and through the packing material that is located at the bottom of the cell. An additional column clean-up module can be added to the output of the extraction cell for cleaning the samples prior to analysis. Additionally, PLE instruments can be interfaced directly with a concentrator to automate the entire sample preparation workflow. This Configuration helps to automate the sample preparation process and improve productivity prior to analysis. Table III lists the key features for the FMS PLE system⁷⁵.

CONCLUSION:

The PLE method was simple, fast and reliable of extraction of grape seed oil. The majority of the Brazilian varieties showed small amounts of α-tocopherol, but as the grape seed a waste material of the industry of the wine and is product high amounts, the extraction of α-tocopherol is a potential to aggregate value to grapes PLE presents good efficiency to take oils from grape seed and the method proposed can be useful in the qualitative and quantitative routine analysis of tocopherols from these oils.The developed PLE method is simple, rapid and accurate for quantitative determination of major volatile compounds.PLE over other extraction approaches, in addition to reduced time/solvent cost, the extract of PLE showed a distinct constituent profile from that of Soxhlet extraction, with significantly improved recovery of active constituents. Pressurized liquid extraction (PLE) is an exhaustive technique used for the extraction of analytes from solid samples. Temperature, pressure, solvent type and volume, and the addition of other reagents notably influence the efficiency of the extraction. The analytical applications of this technique can be improved by coupling with appropriate derivatization reactions.PLE enhances reaction rates and throughput. Fast reaction times can reduce exposure of labile samples to air and light.

ACKNOWLEDGMENT:

The author wants to acknowledge the library of Rungta College of Pharmaceutical Sciences and Research, Kohka-Kurud Road, Bhilai, for providing necessary literature for the compilation of the work. The authors also want to thanks Shri. Santosh Rungta, Chairman, Santosh Rungta Group of Institution for providing necessary facility and infrastructure for the completion of the work.

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