Method optimization for the extraction of chlorogenic acids from coffee parchment: An ecofriendly alternative.

One of the principal byproducts of coffee roasting is the coffee parchment. It is abundant in bioactive substances, including derivatives of chlorogenic acids, which are well-known for their exceptional antioxidant effects. It is advantageous to use environmentally friendly extraction techniques on such residues since it adds value to the entire coffee production process supply chain. The aim of this work was to assess and enhance the ability of non-conventional extraction techniques to extract derivatives of chlorogenic acid from coffee parchment. A central composite design was used to maximize the recovery of those phenolic compounds. The optimized extraction conditions were with 5 min extraction period, at a temperature of 70 °C, and 80% ethanol in the extractor solvent. In this conditions extraction recovery of chlorogenic acids was of 0.8% by the use of microwave-aided extraction (MAE). The optimized conditions are practical, economical, and ecologically friendly method to extract phenolic compounds and, consequently, underscores the potential for sustainable utilization of coffee parchment, offering a valuable contribution to the development of environmentally conscious strategies within the coffee industry.

Comparative assessment of three RNA extraction methods for obtaining high-quality RNA from Candida viswanathii biomass.

Isolating high quality RNA is a limiting factor in molecular analysis, since it is the base for transcriptional studies. The RNA extraction method can directly affect the RNA quality and quantity, as well as, its overall cost. The industrial importance of the yeast genus Candida in several sectors comes from their capacity to produce Lipases. These enzymes are one of the main metabolites produced by some Candida species, and it has been shown that Candida yeast can biodegrade petroleum hydrocarbons and diesel oil from biosurfactants that they can produce, a feature that turns these organisms into potential combatants for bioremediation techniques. Thus, this study aimed to determine an efficient method for isolating high quality RNA from Candida viswanathii biomass. To achieve this aim, three different RNA extraction methods, TRIzol, Hot Acid Phenol, and CTAB (Cetyltrimethylammonium Bromide), were tested. The three tested methods allowed the isolation of high-quality RNA from C. viswanathii biomass and yielded suitable RNA quantity for carrying out RT-qPCR studies. In addition, all methods displayed high sensitivity for the expression analysis of the CvGPH1 gene through RT-qPCR, with TRIzol and CTAB showing the best results and the CTAB method displaying the best cost-benefit ratio (US$0.35/sample).

Development of an aqueous ultrasound-assisted extraction process of bioactive compounds from beet leaves: a proposal for reducing losses and increasing biomass utilization.

BACKGROUND: Red beet plants are cultivated worldwide for the consumption of their roots, generating large amounts of unexploited by-products. In particular, beet leaves (BLs) represent about 50% of the whole plant and are usually discarded as waste. This constitutes not only an economic issue, since multiple resources invested in the production will be wasted, but also an environmental problem because of the pollution associated with their disposal. However, BLs comprise an important source of functional compounds (polyphenols and betalains) that could be recovered from the raw material, representing a sustainable solution for the underutilization of this by-product. This study proposes the recovery of polyphenols and betalains using an aqueous ultrasound-assisted extraction (UAE) process at different powers (35, 50, and 100 W) that was characterized and optimized. RESULTS: UAE significantly enhanced the recovery of bioactive compounds and shortened the time required for extraction in comparison with traditional macerations (35 < 50 < 100 W). During UAE, the temperature of the systems increased as a function of the power applied, favouring the recovery of these phytochemicals. Additionally, a Box-Behnken design and response surface methodology were employed to optimize UAE conditions (90 W ultrasound power, 1:20 solid:liquid ratio, 16 min extraction time), under which the yields were 14.9 mg g-1 (polyphenols), 949.1 µg g-1 (betaxanthins), and 562.2 µg g-1 (betacyanins), consistent with the values predicted by the models. CONCLUSION: This study enabled the development of a green-solvent UAE process that constitutes an effective post-harvest by-products strategy to minimize losses and increase biomass utilization through the recovery of bioactive compounds from BLs, promoting sustainability in the agri-food chain. © 2020 Society of Chemical Industry.

Environmentally Friendly Methods for Flavonoid Extraction from Plant Material: Impact of Their Operating Conditions on Yield and Antioxidant Properties.

The flavonoids are compounds synthesized by plants, and they have properties such as antioxidant, anticancer, anti-inflammatory, and antibacterial, among others. One of the most important bioactive properties of flavonoids is their antioxidant effect. Synthetic antioxidants have side toxic effects whilst natural antioxidants, such as flavonoids from natural sources, have relatively low toxicity. Therefore, it is important to incorporate flavonoids derived from natural sources in several products such as foods, cosmetics, and drugs. For this reason, there is currently a need to extract flavonoids from plant resources. In this review are described the most important parameters involved in the extraction of flavonoids by unconventional methods such as ultrasound, pressurized liquid extraction, mechanochemical, high hydrostatic pressure, supercritical fluid, negative pressure cavitation, intensification of vaporization by decompression to the vacuum, microwave, infrared, pulsed electric field, high-voltage electrical discharges, and enzyme-assisted extraction. There are no unified operation conditions to achieve high yields and purity. Notwithstanding, progress has been achieved in the development of more advanced and environmentally friendly methods of extraction. Although in literature are found important advances, a complete understanding of the extraction process in each of the unconventional techniques is needed to determine the thermodynamic and kinetic mechanisms that govern each of the techniques.

A feasible strategy based on high ultrasound frequency and mass spectrometry for discriminating individuals diagnosed with bipolar disorder and schizophrenia through ionomic profile.

RATIONALE: A viable and accurate method based on high-power ultrasound-assisted microextraction and inductively coupled plasma mass spectrometry was developed to determine metals in human serum from patients with bipolar disorder and schizophrenia. METHODS: A simple and rapid sample preparation method using a cup-horn sonoreactor was developed. The acid concentration of HNO3 (10, 20, and 40% v/v) and HCl (1, 5, 15, and 30% v/v) of the extraction solution, the sonication time (1, 3, 6, and 10 min), and the sonication amplitude (20, 40, 60, and 80%) were evaluated. Cd, Cu, Fe, Li, Pb, and Zn were determined in serum samples from patients with bipolar disorder and schizophrenia, and from healthy controls. Quantitative metal recoveries using the proposed method were compared under the same conditions using an ultrasonic bath, magnetic stirring, and microwave-assisted digestion. RESULTS: Optimum extraction conditions were obtained using HNO3 (40% v/v) + HCl (30% v/v) as the extraction solution with 3 min sonication time and 60% sonication amplitude. Significant differences were observed among the methods compared. On application of the sample preparation method based on high-power ultrasound-assisted microextraction coupled with inductively coupled plasma mass spectrometry, Pb and Cd in all the studied samples were below the limit of detection of our method. Compared with healthy controls, the concentration of Cu, Li, Fe, and Zn was found to be significantly higher for the bipolar disorder group, while these metals and Li were found at a lower level for the group diagnosed with schizophrenia. CONCLUSIONS: Principal component analysis showed a significant separation for the groups studied based on their ionomic profiles after the application of high-power ultrasound-assisted microextraction as a sample preparation strategy.

Optimization and validation of an SBSE-HPLC-FD method using laboratory-made stir bars for fluoxetine determination in human plasma.

Depression is the largest cause of disability worldwide, affecting 350 million people. Notwithstanding that clinical trials demonstrate antidepressants efficacy, the efficient response can vary individually concerning therapeutic dosage. Although important, plasma levels monitoring remains an analytical challenge whereas clean-up and pre-concentration represent critical steps. Therefore, this study aims to develop, optimize and validate a method for fluoxetine determination in human plasma, employing a laboratory-made device consisting of a PDMS stir bar sorptive for extraction, coupled with high-performance liquid chromatography-fluorescence detection (SBSE-HPLC-FD). Optimization involved sorption-desorption steps. For sorption, temperature and time were assessed by factorial and central composite design approaches, taking into account the desirability and the response surface results, with stirring speed also examined. For desorption kinetics and ultrasonic and magnetic stirring mode were evaluated. The proposed method after validation was robust, linear (25.00-1000.00 ng mL-1 , R2 > 0.98) and presented good intra- (RSD 4.18%) and inter-day-assay (RSD 11.60%) precision and accuracy (recovery 109.60%), allowing reliable quantitation without interference. The method was successfully applied to real samples. SBSE-HPLC-FD could represent a feasible alternative with good cost-benefit for low-volume samples and therapeutic drug monitoring, as well as contributing to correlation studies between plasma fluoxetine levels and clinical response, which is still little studied.

Byproduct Generated During the Elaboration Process of Isotonic Beverage as a Natural Source of Bioactive Compounds.

Agro-industrial byproducts are considered good sources of macronutrients and phytochemicals. Fruit and vegetable residues (FVR), obtained after the production of an isotonic beverage, have previously been characterized containing 80% insoluble dietary fibers from total fibers (48.4%), 26% available carbohydrates, 9.5% proteins and 5% lipids. Nevertheless, fruit and vegetables provide phytochemicals which have been related to human health such as phenolic compounds. The loss of specific compounds over the production process is related to their partitioning between fruit and vegetables and byproducts. However, phenolic profile of FVR remains unknown. This work is focused on the evaluation of FVR as a natural source of these bioactive compounds. For this purpose, pressurized liquid extraction (PLE) has been proposed as extraction technique for recovering phenolic compounds from FVR. The experimental variables were temperature and percentage of solvent (ethanol and water). Phenolic compounds extracts were characterized by UPLC-ESI-Q-TOF-MS and a discussion about phenolic and macronutrient interactions was established. Globally, 88 compounds were tentatively identified: phenolic acids (28), flavonoids (32), and other polyphenols (28). The PLE conditions applied yielded different breaking matrix-analyte interactions leading to an increase in the number of compounds. The highest phenolic acids content was achieved with high temperature while lower temperatures were more efficient in extracting flavonoid. By establishing the phenolics profile in food byproducts such as FVR, it is possible to more effectively apply these byproducts as nutraceutical, food or pharmaceutical ingredients. PRACTICAL APPLICATION: Flow diagram of bioactive compounds recovering from isotonic beverage byproduct is proposed using pressurized liquid extraction. The plant-bioactives mechanism relies on fruit and vegetable byproducts changes under different extraction conditions. The obtained extracts can most effectively be applied as nutraceuticals or as ingredients in food or pharmaceutical inputs.

Effect of the type and level of hydration of alcoholic solvents on the simultaneous extraction of oil and chlorogenic acids from sunflower seed press cake.

BACKGROUND: The present study aimed to evaluate the replacement of hexane by alcoholic solvents in oil extraction from sunflower seed press cake. The use of ethanol and isopropanol has important advantages, including low toxicity and good operational safety. Thus, in the present study, solid-liquid extractions were performed in a single stage from 60 to 90 °C and in consecutive extractions in three stages at 90 °C. RESULTS: Solvent hydration negatively affected the extraction of oil but favored the extraction of chlorogenic acids (CAs), especially when ethanol was used. Regarding oxidative stability, the oils extracted using ethanol presented long induction times, which could be related to the high levels of not only CAs and tocopherols, but also phospholipids. CONCLUSION: Alcoholic solvents can be used for extraction to produce sunflower seed oil containing minor compounds that give it greater oxidative stability. In addition, the results obtained using hydrous ethanol showed that this solvent can yield defatted sunflower seed meal with a low content of CAs, enabling future use of the protein fraction. © 2017 Society of Chemical Industry.

A microdevice assisted approach for the preparation, characterization and selection of continuous aqueous two-phase systems: from micro to bench-scale.

Aqueous two-phase systems (ATPS) have emerged as an alternative strategy for the recovery and purification of a wide variety of biological products. Typical process development requires a large screening of experimental conditions towards industrial adoption where continuous processes are preferred. In this work, it was proved that under certain flow conditions, ATPS could be formed continuously inside a microchannel, starting from stocks of phase components. Staggered herringbone chaotic micromixers included within the device sequentially and rapidly prepare two-phase systems across an entire range of useful phase compositions. Two-phase diagrams (binodal curves) were easily plotted using the cloud-point method for systems of different components and compared with previously reported curves for each system, proving that phase formation inside the device correlated with the previously reported diagrams. A proof of concept for sample partitioning in such a microdevice was performed with two different experimental models: BSA and red blood cells. Finally, the microdevice was employed to obtain information about the recovery and partition coefficient of invertase from a real complex mixture of proteins (yeast extract) to design a process for the recovery of the enzyme selecting a suitable system and composition to perform the process at bench-scale.

Multivariate optimization of headspace-GC for the determination of monoaromatic compounds (benzene, toluene, ethylbenzene, and xylenes) in waters and wastewaters.

The objective of this study was to optimize, by employing a central composite rotatable design, and validate an analytical method to detect and quantify monoaromatic compounds (benzene, toluene, ethylbenzene, and xylenes) in waters and wastewaters by using headspace extraction followed by GC coupled with photoionization detection. The extraction parameters optimized were: salinity, sample volume, incubation time, and extraction temperature. The results revealed that the sample volume was the most significant parameter in the extraction process, whereas the salinity effect was negligible, which extends the applicability of the analytical method to waters with different salinities. Finally, the studied method was very selective and, at the optimal extraction conditions (15 mL sample volume, 15 min incubation time, and temperature of 70°C), presented excellent repeatability (<4%), linearity (R > 0.999 for each compound), and sensitivity, since very low LODs (0.13-0.48 µg/L) and LOQs (0.43-1.61 µg/L) were achieved.

Repetitive use of polyacrylamide gels for the analysis and purification of oligonucleotides.

The repetitive use of polyacrylamide slab gels for the analysis of oligonucleotides by ultraviolet (UV) shadowing across the complete electrophoresis cassette is addressed. Visualization of the samples is possible by preparing the gel between a quartz plate and a highly sensitive glass fluorescent plate whose preparation is described here. Within a working day, gels can be reloaded up to five times with no detriment to the resolution of the oligonucleotide bands. Taking advantage of this detection approach, we also devised a strategy to reuse gels in the purification of oligonucleotides, combining electroelution and subsequent concentration of the samples using centrifugal filter units.

The enhancement of antioxidant compounds extracted from Thymus vulgaris using enzymes and the effect of extracting solvent.

We evaluate the total phenolic compounds (TPC) content and the antioxidant activity (AA) of extracts obtained from ground fresh thyme (FT) and depleted thyme (DT), a by-product of the process of essential oil extraction. In addition, enzymatic treatments were evaluated to improve the extraction yields of polyphenolic compounds from thyme. Extractions were performed using several solvents as methanol, ethanol, and water. Enzymes were applied prior to extraction or during the extraction process. The best results were obtained using a mixture of methanol and water, resulting in 2790 and 220 mg Gallic acid equivalent (GAE)/L of TPC for FT and DT, respectively. A similar result was observed for AA. With regard to enzymatic treatment, application of Grindamyl CA 150 enzyme as a pre-treatment resulted in the production of an extract from DT with 614 mg TE (trolox equivalent)/L of AA, 70% more than the control, and an AA of 621 mg TE/L (74% more than the control sample) was obtained using Grindamyl CA 150 during the extraction process. These results suggest that enzymatic treatment is an interesting alternative for producing antioxidant extracts from DT.

Microwave-assisted methanolysis of green coffee oil.

Optimisation of a microwave-assisted methanolysis was performed to obtain cafestol and kahweol directly from green coffee oil (Coffea arabica). A two-factor (the methanolysis period and temperature), three-level, factorial experimental design (3(2)) was adopted. The methanolysis procedure was performed under microwave irradiation, using closed vessel and accurate fast responding internal fibre-optic temperature probe. The effects on the responses were measured by HPLC. After 3 min of microwave irradiation (hold time) at 100°C, with 500 mg of green coffee oil, a yield higher than 99% was obtained. The yield of this reaction is 26% after 2h when working under conventional heating. The methods described in the literature lead to long reaction times, poor yields and formation of side products. The microwave-assisted technique proved to be faster, avoided undesired side products and gave better conversion, when compared to conventional heating process.

Rotating disk sorbent extraction for pre-concentration of chromogenic organic compounds and direct determination by solid phase spectrophotometry.

A novel and very simple microextraction approach for pre-concentration and direct solid phase spectrophotometric measurement has been developed for the determination of chromogenic analytes. The model analyte to assess this approach was the chromophore malachite green (MG). The analyte was extracted from water samples onto a small rotating disk made of Teflon containing a sorbent phase of polydimethylsiloxane (PDMS) on one of its surfaces. We refer to the extraction procedure as rotating disk sorptive extraction (RDSE). After extraction, the sorbent phase with the concentrated analyte was separated from the Teflon disk and used directly for MG determination by solid phase spectrophotometry at 624 nm, without the necessity of a desorption step. Chemical and extraction variables such as concentration of sodium sulfate, pH, disk rotational velocity, extraction time, and temperature were studied in order to establish the best conditions for extraction. Under optimum conditions, the extraction of MG was carried out in 18 min and 90 min, for sample volumes of 100mL or 1000 mL, respectively. The detection limit, based on three times the standard deviation of the blank phase (3σ(b)), was 1.4 µg L⁻¹ and the repeatability, expressed as relative standard deviation (RSD), for 20 µg L⁻¹ MG was 8.1%. This study also applied the method to real samples, obtaining quantitative recovery (mean recovery of 99.3%). The PDMS phases could be reused after desorbing the MG into methanol for 3h. Replacement of the PDMS film onto the disk is very easy and low cost.

Simple determination of 40 organophosphate pesticides in raw wool using microwave-assisted extraction and GC-FPD analysis.

A validated analytical method for the multiresidue analysis of 40 organophosphate pesticides (OPs) and conversion products in raw wool has been developed. The method is based on the selective microwave-assisted extraction (MAE) of raw wool with acetonitrile and analysis of extracts by gas chromatography-flame photometric detector. The optimum MAE conditions were 20 min duration at 80 °C with 30 mL of acetonitrile per gram of wool. A validation study was performed according to the European SANCO guidelines 10684/2009. Limits of detection and quantification for all pesticides tested were from 0.01 to 0.2 mg/kg and from 0.2 to 1.0 mg/kg, respectively. The average recoveries of pesticides spiked at different levels were in the range of 70-120% with relative standard deviations of ≤ 20%. The extraction performance was compared to the one obtained with a reference Soxhlet extraction. The method was also applied in the analysis of real wool (after field application) samples.

Use of multivariate statistical techniques to optimize the simultaneous separation of 13 phenolic compounds from extra-virgin olive oil by capillary electrophoresis.

Characterization of phenolic compounds in olive oil has not been achieved as yet, owing to the complexities of their chemical structures and analytical matrix. The aim of this work is to optimize and validate a method for simultaneous separation and quantification of 13 phenolic compounds from extra-virgin olive oil: tyrosol, hydroxytyrosol, oleuropein glycoside, ferrulic acid, p-coumaric acid, cinnamic acid, p-hydroxybenzoic acid, gallic acid, caffeic acid, luteolin, apigenin, vanillic acid and 3,4-dihydroxybenzoic acid. A statistical central composite design, response surface analysis and the simultaneous optimization method of Derringer and Suich were used to separate all the peaks. These multivariate procedures were efficient in determining the optimal separation condition, using five peak-pair resolutions and runtime as responses. The optimized method employed a fused-silica capillary of 50 µm i.d.× 60 cm effective length with extended light path, 50 mmol L(-1) boric acid electrolyte, 10.2 pH, 25°C, injection of 50 mbar for 25s with application of reverse voltage (-30 kV for 5s) before setting the running voltage (+30 kV) with detection at 210 nm and a run time of 12 min. Peak resolutions are found to be very sensitive to pH values outside the 10.15-10.25 range but acceptable electropherograms can be obtained for a wide range of boric acid concentrations within this pH interval.

Hollow fiber-based liquid-phase microextraction (HF-LPME) of isradipine and its main metabolite followed by chiral HPLC analysis: application to an in vitro biotransformation study.

An enantioselective liquid chromatographic method using two-phase hollow fiber liquid-phase microextraction (HF-LPME-HPLC) was developed for the determination of isradipine (ISR) enantiomers and its main metabolite (pyridine derivative of isradipine, PDI) in microsomal fractions isolated from rat liver. The analytes were extracted from 1 mL of microsomal medium using a two-phase HF-LPME procedure with hexyl acetate as the acceptor phase, 30 min of extraction, and sample agitation at 1,500 rpm. For the first time, ISR enantiomers and PDI were resolved. For this separation, a Chiralpak(®) AD column with hexane/2-propanol/ethanol (94:04:02, v/v/v) as the mobile phase at a flow rate of 1.5 mL min(-1) was used. The column was kept at 23 ± 2 °C. The drug and metabolite detection was performed at 325 nm and the internal standard oxybutynin was detected at 225 nm. The recoveries were 23% for PDI and 19% for each ISR enantiomer. The method presented quantification limits (LOQ) of 50 ng mL(-1) and was linear over the concentration range of 50-5,000 and 50-2,500 ng mL(-1) for PDI and each ISR enantiomer, respectively. The validated method was employed to an in vitro biotransformation study of ISR using rat liver microsomal fraction showing that (+)-(S)-ISR is preferentially biotransformed.

Simultaneous determination of rosiglitazone and its metabolites in rat liver microsomal fraction using hollow-fiber liquid-phase microextraction for sample preparation.

A three-phase hollow-fiber liquid-phase microextraction method for the analysis of rosiglitazone and its metabolites N-desmethyl rosiglitazone and ρ-hydroxy rosiglitazone in microsomal preparations is described for the first time. The drug and metabolites HPLC determination was carried out using an X-Terra RP-18 column, at 22°C. The mobile phase was composed of water, acetonitrile and acetic acid (85:15:0.5, v/v/v) and the detection was performed at 245 nm. The hollow-fiber liquid-phase microextraction procedure was optimized using multifactorial experiments and the following optimal condition was established: sample agitation at 1750 rpm, extraction for 30 min, hydrochloric acid 0.01 mol/L as acceptor phase, 1-octanol as organic phase, and donor phase pH adjustment to 8.0. The recovery rates, obtained by using 1 mL of microsomal preparation, were 47-70%. The method presented LOQs of 50 ng/mL and it was linear over the concentration range of 50-6000 ng/mL, with correlation coefficients (r) higher than 0.9960, for all analytes. The validated method was employed to study the in vitro biotransformation of rosiglitazone using rat liver microsomal fraction.

Hollow fiber-based liquid phase microextraction with factorial design optimization and gas chromatography-tandem mass spectrometry for determination of cannabinoids in human hair.

A new method, based on hollow fiber liquid-phase microextraction (HF-LPME) and gas chromatography-tandem mass spectrometry (GC-MSMS), was developed for determination of Delta(9)-tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN) in samples of human hair. Since hair is a solid matrix, the samples were subjected to alkaline digestion using NaOH. The aqueous solutions obtained were extracted using a 6cm polypropylene fiber (600microm i.d., 200microm wall thickness, 0.2microm pore size) for each extraction. A 2(5-1) fractional factorial design for screening, and a central composite design for optimization of significant variables, was applied during development of the extraction method. The variables evaluated were the type of extraction solvent, pH, stirring speed, extraction time, and acceptor phase volume. The optimized conditions for the proposed extraction procedure were 10mg of hair sample; 20microL of butyl acetate; aqueous (pH 14) donor phase containing 6.8% NaCl; 600rpm stirring speed; 20min extraction time. A linear response was obtained in the ranges 1-500pgmg(-1) (CBD and CBN) and 20-500pgmg(-1) (THC), with regression coefficients >0.99. Precision, determined as the relative standard deviation, was 3.3-8.9% (intra-day) and 4.4-13.7% (inter-day). Absolute recoveries varied in the ranges 4.4-4.8% (CBD), 7.6-8.9% (THC) and 7.7-8.2% (CBN). Limits of detection (LOD, S/N=3) and quantification (LOQ, S/N=10) were 0.5-15pgmg(-1) and 1-20pgmg(-1), respectively. The method was successfully used to determine CBD, THC and CBN in hair samples from patients in a drug dependency rehabilitation center. Concentrations varied in the ranges 1-18pgmg(-1) (CBD), 20-232pgmg(-1) (THC) and 9-107pgmg(-1) (CBN), confirming the suitability of the method for monitoring studies.

Two-step liquid-phase microextraction and high-performance liquid chromatography for the simultaneous analysis of the enantiomers of mefloquine and its main metabolite carboxymefloquine in plasma.

A method for the simultaneous analysis of the enantiomers of mefloquine (MQ) and its main metabolite carboxymefloquine (CMQ) in plasma is described for the first time. The assay involves two-step liquid-phase microextraction (LPME) and enantioselective high-performance liquid chromatography. In the first LPME step, the enantiomers of MQ were extracted from an alkalinized sample through a thin layer of di-n-hexyl ether immobilized in the pores of the hollow fiber and into 0.01 M perchloric acid as acceptor solution. In the second LPME step, the same sample was acidified to enable the extraction of CMQ using the same organic solvent and 0.05 M sodium hydroxide as acceptor phase. The analytes were resolved on a Chirobiotic T column in the polar-organic mode of elution and detected at 285 nm. The recovery rates from 1 mL of plasma were in the range 35-38%. The method presented limits of quantification of 50 ng/mL for all analytes and was linear up to 1,500 and 3,000 ng/mL for the enantiomers of MQ and CMQ, respectively. The plasmatic concentrations of (+)-(RS)-MQ were higher than those of (-)-(SR)-MQ after oral administration of the racemic drug to rats.