Rapid identification of cervus antlers by species-specific PCR assay

Abstract

A rapid PCR technology was developed to differentiate Cervus antlers species and adulteration based on the difference in mitochondrial genome. Three specifically designed primer sets were confirmed to have high inter-species specificity and good intra-species stability. Limits of detection were estimated to be 1?ng of genomes for reindeer and 10?ng for the other species. Especially, when the mixture of Cervus antlers and reindeer or sambar was assayed, these primer sets still exhibited strong capability of differentiation but not the conventional COI barcoding. By using the newly developed approach, five batches out of fourteen commercial Cervus antler products were identified to be fake products made from reindeer antlers. It has shown its good potential to be extensively applied in the identification of counterfeits or adulterates of Cornu Chinese medicines for their pulverized and processed form, and even the traditional Chinese patent medicines composed of these species.     

Quantitative Determination and Variation Trends of Six Alkaloids in Crude and Processed Corydalis Yanhusuo

A convenient, sensitive, and accurate analytical method using reversed-phase high-performance liquid chromatography coupled with ultraviolet-visible detection was investigated for the determination of 6 alkaloids (tetrahydrocoptisine, protopine, tetrahydropalmatine, coptisine, corydaline, and palmatine) in crude and processed samples of corydalis yanhusuo. The results revealed that the content of the alkaloids in crude corydalis yanhusuo samples varied from those of the processed samples. The concentrations of tetrahydrocoptisine, protopine, tetrahydropalmatine, coptisine, and corydaline were reduced after processing; however, the concentration of palmatine increased after processing. This method was established using a C₁₈ column (250 mm × 4.6 mm, 5 µm). The mobile phase, a mixture (19:81) of acetonitrile and a 0.6% solution of acetic acid (pH 3.5, adjusted using triethylamine), was used to elute the target compounds in the isocratic elution mode. The solvent flow rate was 1.0 mL/min, the column temperature was 35°C, and the detection wavelength was 280 nm. All of the calibration curves exhibited good linearity with R >0.9999. The inter- and intra-day precisions for all of the investigated components ranged from 0.10–2.64% RSD, and the recoveries were in the range of 97.3–102.5%. The validated method provides a new solution for analyzing the contents of crude and processed samples of corydalis yanhusuo.     

LC-DAD-ESI-MS Characterization of Carbohydrates Using a New Labeling Reagent

A sensitive and selective liquid chromatographic tandem mass spectrometric (LC–MS–MS) method was developed for simultaneous identification and quantification of tamsulosin and dutasteride in human plasma, which was well applied to clinical study. The method was based on liquid–liquid extraction, followed by an LC procedure with a Gemini C-18, 50 mm × 2.0 mm (3 μm) column and using methanol:ammonium formate (97:3, v/v) as the mobile phase. Protonated ions formed by a turbo ionspray in positive mode were used to detect analytes and internal standard. MS–MS detection was by monitoring the fragmentation of 409.1 → 228.1 (m/z) for tamsulosin, 529.3 → 461.3 (m/z) for dutasteride and 373.2 → 305.3 (m/z) for finasteride (IS) on a triple quadrupole mass spectrometer. The lower limit of quantification for both tamsulosin and dutasteride was 1 ng mL⁻¹. The proposed method enables the unambiguous identification and quantification of tamsulosin and dutasteride for clinical drug monitoring.

     

Identification and Quantification of Heterologous Compounds Parthenin and Organic Acids in Parthenium Hysterophorus L. Using HPLC-PDA-MS-MS

Parthenium hysterophorus L., is an obnoxious weed known for its environmental health hazards and medicinal uses. These characteristics are due to presence of sesquiterpene lactones and organic acids; therefore a rapid and sensitive analytical procedure using HPLC-PDA-MS-MS was developed and optimized for separation, identification, and quantification of parthenin and six organic acids. Separation and characterization of compounds was achieved on a RP-C18 column with 1% acetic acid in water (A) and acetonitrile (B) as a mobile phase at a flow rate of 0.6 mL min^?1 and by matching their UV and mass spectra with reference compounds. Six organic acids (ferulic acid, 0.1 mg g^?1 to coumaric acid, 13.6 mg g^?1) and parthenin (27.4 mg g^?1) were characterized within 26 minutes of chromatographic separation in plant extract. The calibration curves are linear with correlation coefficients from 0.985 to 0.998, limit of detection and quantification ranged between 1.0 µg mL^?1 (anisic acid) to 2.2 µg mL^?1 (parthenin) and 2.5 µg mL^?1 (coumaric acid) to 5.2 µ g mL^?1 (parthenin) and recovery ranged between 90.9% to 97.3%. To the best of our knowledge this is the first report for the simultaneous separation of parthenin and organic acids. The method is applicable for screening of commercial crops, medicinal plants, and their products which might be mixed with P. hysterophorus during harvesting period.     

Identification and quantification of domoic acid by UHPLC/QTOF tandem mass spectrometry,with simultaneous identification of non-target photodegradation products

Amnesic shellfish poisoning is a potentially lethal human toxic syndrome which is caused by domoic acid (DA), a neurotoxin produced by marine phytoplankton, principally from Pseudonitzschia genus. In this report, a method to identify and quantify the DA toxin, with simultaneous identification of its photodegradation products, has been developed. It uses an ultra high performance liquid chromatography coupled to a quadrupole-time-of-flight tandem mass spectrometer (UHPLC–QTOF) after solid-phase extraction (SPE). An unambiguous identification of DA was carried out by considering both the retention time of DA in UHPLC and the exact mass of protonated DA molecule ([M + H]⁺ = 312.1447 m/z) and of the most intense fragment ion (m/z 266.1391). The quantification was conducted using protonated DA molecule with protonated Glafenin as internal standard, obtaining a limits of detection of 0.75 µg L^?1. Large screening with UHPLC–QTOF could also give structural information about degradation products of DA present in samples after UV-irradiation. This method was applied for the determination of DA in complex liquid samples after SPE and is applicable for environmental monitoring of this toxic substance in the aquatic environment.     

Optimization of Sample Preparation of Carrot-Fruit Juice for Determination of Antimony,Arsenic, and Selenium by Hydride Generation-Inductively Coupled Plasma Optical Emission Spectrometry

Sample preparation procedures for the determination of As, Sb, and Se in carrot-fruit juice by hydride generation inductively coupled plasma optical emission spectrometry (HG-ICP OES) were examined. The applicability of a partial decomposition using aqua regia and simple dilution with a 2% (v/v) HNO₃ solution were tested and compared to a traditional treatment based on the wet digestion with a HNO₃/H₂O₂ mixture. The pre-reduction and hydride generation reaction conditions were evaluated. Under the optimal conditions, the hydrides were produced in the reaction of an acidified sample with NaBH₄ after pre-reduction with ascorbic acid [0.5% (m/v)] and KI [0.5% (m/v)] in 3 mol L^?1 HCl for total As and Sb, and boiling with HCl (6 mol L^?1) for total Se. The best results were obtained for the aqua regia procedure, resulting in limits of detection (LODs) between 1.2–2.4 ng g^?1 in the samples and recoveries from 90.9% to 109.1%. The method was successfully applied (without matrix effects) for the determination of As in dense mousse and pulp juice samples and for Sb in pulp juices. Standard solutions, processed in the same way as samples, were used for the calibration. Undecomposed matrix constituents strongly influenced Se; hence this element was determined using the method of standard addition. Concentrations of studied elements in analyzed products were at the trace level, that is, 6–32 ng g^?1, 4–10 ng g^?1, and 4–13 ng g^?1 for Se, As, and Sb, respectively.     

Evaluation of seasonal chemical composition,antibacterial, antioxidant and anticholinesterase activity of essential oil from Eugenia brasiliensis Lam.

This study describes the seasonal composition and the antibacterial, antioxidant and anticholinesterase activity of the essential oil from Eugenia brasiliensis leaves. Analysis by using GC allowed the identification of 40 compounds. It was observed that the monoterpenes varied more (42%) than the sesquiterpenes (14%), and that the monoterpene hydrocarbons suffered the greatest variation throughout the year (64%). Major compounds were spathulenol in the spring (16.02 ± 0.44%) and summer (18.17 ± 0.41%), τ-cadinol in the autumn (12.83 ± 0.03%) and α-pinene (15.94 ± 0.58%) in the winter. Essential oils were tested for their antibacterial activity, and the best result was obtained from the autumn oil, with MIC = 500 μg mL^? 1 against Staphylococcus saprophyticus and Pseudomonas aeruginosa. Antioxidant activity was evaluated using DPPH, lipid peroxidation and iron-reducing power assays, as well as the anticholinesterase activity. Both tests showed a weak performance of the essential oils.     

Quality evaluation of raw and processed Crataegi Fructus by color measurement and fingerprint analysis

Crataegi Fructus and its processed products have been used as a traditional medicine for a long time, and numerous active components are responsible for their curative effects. However, a comprehensive and fast method for the quality control of its processed products is still lacking. In this study, two analytical methods based on color measurements and fingerprint analysis are established. In the color measurements, the color values of the peel and flesh of Crataegi Fructus were evaluated spectrophotometrically. Based on the results, a color reference range was established using percentiles, and the standard color difference value was established using the median color values. Then, the color values of Crataegi Fructus and its processed products were analyzed using Bayes linear discriminant analysis and mathematical functions were built in order to predict the degree of processing. Moreover, high‐performance liquid chromatography fingerprint analysis was performed on a Hibar C₁₈ column, and a high‐performance liquid chromatography fingerprint pattern was obtained, from which nine peaks were identified. Chemometric methods were successfully applied to differentiate raw and processed Crataegi Fructus.     

LC-ESI-MS/MS Method for Simultaneous Determination of Triterpenoid Glycosides and Aglycones in Centella asiatica L.

A new and rapid method for simultaneous identification and estimation of bioactive triterpenoid glycosides [asiaticoside (AS) and madecassoside (MS)] and their aglycones [asiatic acid (AA) and madecassic acid (MA)] in Centella asiatica was developed by using high-performance liquid chromatography (HPLC) coupled with triple-quadrupole mass spectrometry (MS/MS). Estimation was based on multiple reaction monitoring (MRM) using the precursor → product ion combination for determination of four analytes using Alltima C18 column (50 × 4.6 mm, 3 µm). An electrospray ionization (ESI) tandem interface in positive mode was employed prior to mass-spectrometric detection. The method was subjected to a thorough validation procedure in terms of linearity, limit of detection (LOD) and quantification (LOQ), accuracy, and precision. Six-point calibration curves were linear in the range of 50–500 ng mL⁻¹ for AS and MS, and 25–250 ng mL⁻¹ for AA and MA, with excellent linearity (R ² > 0.98). With the optimized conditions, the four analytes were detected accurately within 10 min. LOD and LOQ ranged from 2.5 to 5 and 10 to 15 ng mL⁻¹, respectively. Method accuracy in terms of average recoveries of all four analytes ranged between 98.61 and 102.85 % at three spiking levels with intra- and interday precision relative standard deviation (RSD, %) of 1.01–4.62 and 1.13–4.16, respectively. The new method was successfully applied to estimate the concentration of these four bioactive compounds in extracts of C. asiatica prepared by nonpolar-to-polar solvents.

     

Semi-preparative HPLC purification of δ-tocotrienol (δ-T3) from Elaeis guineensis Jacq. and Bixa orellana L. and evaluation of its in vitro anticancer activity in human A375 melanoma cells

In this work, we report a rapid and convenient HPLC-UV-DAD method for the isolation of δ-T3 on semi-preparative scale from two different vitamin E rich processed, commercially available products obtained from the fruits of Elaeis guineensis Jacq. (oil palm) and from the seeds of Bixa orellana L. (achiote tree). Chromatography was run using reverse phase (RP) C-18 columns and HPLC-grade acetonitrile as mobile phase. The purity of the isolated δ-T3, assessed by GC–MS and ¹H NMR was above 98%. The δ-T3 cytotoxic activity found in vitro against the proliferation of human A375 melanoma cells compared to that of the other δ-T3 free tocols mixture suggest its primary role in the experimental anticancer activity observed for palm oil derived products. Taken altogether, the results of this study highlight the importance of the application of suitable purification systems for the preparations of tocotrienols prior to their experimental or clinical testing.     

Fast HPLC Method for Determination of Fenoxycarb and Permethrin in Antiparasitic Veterinary Shampoo Using Fused-Core Column

A new and fast high-performance liquid chromatography (HPLC) method using technology of fused-core columns for separation of fenoxycarb and cis-, trans-permethrin has been developed and used for their determination in antiparasitic veterinary shampoo. Separation of insecticides and internal standard sudan II was achieved on the fused-core column Ascentis Express RP-Amide (100 × 3.0 mm), particle size 2.7 μm, with mobile phase acetonitrile/water (55:45, v/v) at a flow rate of 1.0 mL min⁻¹ and at temperature 60 °C. The detection wavelength of detector was set at 225 nm for both compounds and internal standard sudan II. Under the optimum chromatographic conditions standard calibration curves were measured with good linearity [r ² = 0.99991 for fenoxycarb, r ² = 0.99987 for trans-permethrin, and r ² = 0.99984 for cis-permethrin (n = 8)]. Commercial samples of antiparasitic veterinary shampoo were extracted with ethanol in ultrasound bath for 5 min. A 2-μL sample volume of the filtered solution was directly injected into the HPLC system. Accuracy of the method defined as a mean recovery of insecticides from shampoo matrix was in the range 100.43–103.85 % for both insecticides.

     

Stability-Indicating Chromatographic Methods for Simultaneous Determination of Mosapride and Pantoprazole in Pharmaceutical Dosage Form and Plasma Samples

Simple, sensitive, selective, precise, and stability-indicating thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) methods for the determination of mosapride and pantoprazole in pharmaceutical tablets were developed and validated as per the International Conference on Harmonization guidelines. The TLC method employs aluminum TLC plates precoated with silica gel 60F₂₅₄ as the stationary phase and ethyl acetate/methanol/toluene (4:1:2, v/v/v) as the mobile phase to give compact spots for mosapride (R _f 0.73) and pantoprazole (R _f 0.45) separated from their degradation products; the chromatogram was scanned at 276 nm. The HPLC method utilizes a C18 column and a mobile phase consisting of acetonitrile/methanol/20 mM ammonium acetate (4:2:4, v/v/v) at a flow rate of 1.0 mL min⁻¹ for the separation of mosapride (t _R 11.4) and pantoprazole (t _R 4.4) from their degradation products. Quantitation was achieved with UV detection at 280 nm. The same HPLC method was successfully used in performing calibrations in lower concentration ranges for both drugs in human plasma using ezetimibe as internal standard. The methods were validated in terms of accuracy, precision, linearity, limits of detection, and limits of quantification. Mosapride and pantoprazole were exposed to acid hydrolysis and then analyzed by the proposed methods. As the methods could effectively separate the drugs from their degradation products, these techniques can be employed as stability-indicating methods that have been successively applied to pharmaceutical formulations without interference from the excipients. Moreover the HPLC method was successfully used in the determination of both drugs in spiked human plasma.

     

Separation of Lactoferrin from Human Saliva Using Monolithic Disc

In this study, we optimized method for the isolation and detection of lactoferrin from human saliva using 3 mm short monolithic disc. We optimized the conditions for separation as flow rate 4 mL min⁻¹ and ionic strength of effluent as 2 M·NaCl. We estimated limit of detection of whole method, which was hyphenated to the Bradford’s assay, down to 100 ng mL⁻¹. The purity of the isolated fractions was verified by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and recovery of isolation was found to be 51 % using minimally processed sample of saliva. Further, we tested the optimized method on group of healthy volunteers (n = 7). We were able to distinguish between the healthy subjects and subject suffering from celiac disease, which reported at least 2.5× higher level of lactoferrin in comparison to healthy ones. The results were correlated with standard enzyme-linked immunosorbent assay (ELISA) kit with obtained correlation coefficient R ² = 0.8446. Analysis of lactoferrin in saliva by monolithic disc and subsequent offline photometric detection is faster and cheaper method compared to ELISA commercial kit. The total analysis of one sample takes <20 min.

     

Thermal compatibility between hydroquinone and retinoic acid in pharmaceutical formulations

Thermogravimetry (TG) and differential scanning calorimetry (DSC) are used in pharmaceutical studies for characterization of drugs, purity, compatibility of formulations, identification of polymorphism, evaluation of stability, and thermal decomposition of drugs and pharmaceutical formulations. Hydroquinone (HQ) and products containing HQ have been widely used as depigmentation agents for lightening the skin. Retinoids are compounds that have the basic core structure of vitamin A and its oxidized metabolites, or synthetic compounds that share similar mechanisms of action as naturally occurring retinoids. Depigmentants and excipients were analyzed by TG and DSC. The dynamic thermogravimetric curves were obtained on a SHIMADZU thermobalance, model DTG-60, using an alumina crucible, at the heating rate of 10 °C min^?1, in the temperature range of 25–900 °C, under an atmosphere of nitrogen at 50 mL min^?1. The sample's mass was 10 ± 0.05 mg. The DSC curves were obtained using Shimadzu calorimeter, model DSC-60, using aluminum crucible, at the heating rate of 10 °C min^?1, in the temperature range of 25–400 °C. The thermogravimetric and calorimetric curves were analyzed using TASYS software SHIMADZU. In this study were found the interaction between retinoic acid (RA) and the following excipients: cetyl alcohol(CA), cetostearyl alcohol (CTA), glycerin(GLY), and dipropylene glycol (DPG), and that between HQ and the excipient, DPG. Therefore, additional studies are necessary to evaluate final formulations. Thermal analysis is an effective and reliable technique that can be used in the control of raw materials and pharmaceutical products, and for evaluating their employment potential in the development and characterization of products.     

A new triterpenoid saponin from Gleditsia sinensis and its antiproliferative activity

Chemical investigation of the anomalous fruits of Gleditsia sinensis led to the isolation and identification of a new triterpenoid saponin, 3-O-β-D-xylopyranosyl-(1 → 2)-α-L-arabinopyranosyl-(1 → 6)-β-D-glucopyranosyl oleanolic acid 28-O-β-D-xylopyranosyl-(1 → 4)-α-L-rhamnopyrano--syl-(1 → 4)-β-D-xylopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 3)-β-D-glucopyranosyl ester (1), along with other nine known compounds (2–10). All the isolates from this species were reported for the first time. The structure of Compound 1 was determined by a detailed analysis using various analytical techniques, including 1D and 2D NMR. In vitro antiproliferative activities of Compound 1 on MCF-7 and Hep-G2 tumor cell lines were evaluated. IC₅₀ values against the two cell lines were 9.5 and 11.6 μM, respectively.     

Study of Forced Degradation Behaviour of Brinzolamide Using LC–ESI–Q-TOF and In Silico Toxicity Prediction

Brinzolamide (BZA) is a topical ophthalmic drug which is generally used to lower the intraocular pressure during glaucoma. It was subjected to forced degradation studies under hydrolytic (acidic, basic), oxidative, photolytic and thermal stress conditions; the drug degraded significantly in hydrolytic and oxidative conditions, leading to the formation of seven degradation products in total. It was stable on exposure to light and dry heat in the solid state. An ultra-performance liquid chromatography (UPLC) method was developed on a Waters CSH phenyl hexyl column (100 × 2.1 mm, 1.7 µm), using gradient elution of 0.1 % formic acid and methanol as mobile phase. The method was extended to quadrupole time-of-flight mass spectrometry (Q-TOF–MS) for the structural characterisation. All degradation products were comprehensively characterised by UHPLC–ESI/MS/MS experiments. The most probable mechanisms for the formation of degradation products were also proposed. In silico toxicity of the drug and its degradation products was determined using TOPKAT toxicity prediction software.

     

TLC Determination of Amitriptyline HCl,Trifluoperazine HCl,Risperidone and Alprazolam in Pharmaceutical Products

A simple, sensitive, and precise thin layer chromatographic (TLC) method for simultaneous analysis of psychopharmacological drugs like amitriptyline HCl, trifluoperazine HCl, risperidone and alprazolam in their single dosage forms has been developed, validated, and used for determination of the compounds in commercial pharmaceutical products. The TLC separation was carried out on Merck TLC aluminium sheets of silica gel 60 F254 using carbon tetrachloride:acetone:triethylamine (8:2:0.3, v/v/v), as mobile phase. Densitometric measurements of their spots were achieved at 250 nm over the concentration range for amitriptyline HCl (50–1,200 ng spot⁻¹), trifluoperazine HCl (50–1,200 ng spot⁻¹), risperidone (100–2,400 ng spot⁻¹) and alprazolam (25–600 ng spot⁻¹). Limit of detection (LOD) for amitriptyline HCl (20 ng spot⁻¹), trifluoperazine HCl (20 ng spot⁻¹), risperidone (40 ng spot⁻¹) and alprazolam (5 ng spot⁻¹) was obtained. The study showed that TLC was sensitive and selective for determination of amitriptyline HCl, trifluoperazine HCl, risperidone and alprazolam using a single mobile phase. This proposed method is able for simultaneous determination of psychopharmacological drugs and also applicable for analysis of pharmaceutical formulations.

     

LC-DAD and LC–MS–MS Analysis of Piperidine Alkaloids of Lobelia inflata L. (In Vitro and In Vivo)

An LC-DAD method was developed for determination of lobeline from in vitro and in vivo cultures of Lobelia inflata. Samples were extracted with 0.1 N HCl–acetonitrile (1:1, v/v), and purified by solid-phase extraction. Optimized conditions resulted in high recovery. LC separations were performed on an Eurosphere C8 reversed-phase column using 30:70 (v/v) acetonitrile–0.1% trifluoroacetic acid as a mobile phase. Quantitative determination of lobeline was performed by external standard method at 250 nm, in the range of 2.4–80 μg mL⁻¹. Validation studies proved that the repeatability of the method was good and the recovery was satisfactory. In vitro organized cultures contained considerable amount of lobeline (herb: 175 μg g⁻¹, root: 100 μg g⁻¹). When these cultures were transplanted into the open field, the lobeline content increased significantly (herb: 323 μg g⁻¹, root: 833 μg g⁻¹). Plants obtained from seed propagation contained 382 μg g⁻¹ lobeline in the herb. For direct characterization of di-substituted piperidine alkaloids in extracts of L. inflata, tandem mass spectrometric method was developed using electrospray ionization. Analysis was performed in the positive ion mode on a triple quadropole LC–MS system. LC separations were achieved on Eurosphere C8 column with a modified mobile phase (acetonitrile–30 mM ammonium formate, pH 2.80) to ensure proper molecular ionization. The identification and structural elucidation of the alkaloids were performed by comparing their changes in molecular mass (ΔM), full-scan MS–MS spectra with those of lobeline, norlobelanine and lobelanidine. These alkaloids and ten other derivatives were identified in the plant extracts. Three piperidine alkaloids were reported in L. inflata for the first time.

     

Chemometrics-assisted cross injection analysis for simultaneous determination of phosphate and silicate

This work presents a novel method for simultaneous spectrophotometric determination of phosphate and silicate by using a cross injection analysis (CIA) coupled with the use of partial least squares (PLS) for data evaluation. The detection principle is based on the well-known ‘molybdenum blue’ method. The molybdate ions in the presence of stannous chloride in acidic medium give phosphomolybdenum blue and silicomolybdenum blue as products. In this work, all the liquids, including sample and reagents were simultaneously introduced into a CIA platform by using two peristaltic pumps for controlling the x-channel and y-channel flow which was automatically manipulated by using in-house control board. Crossflow provides sufficient mixing inside the platform prior detection of the absorption spectra of blue complexes in the wavelength of 400–900 nm. Since spectra of the blue colour product of phosphate and silicate are resemblant, these two analytes therefore reciprocally interfere with one another. This results in difficulty in simultaneous analysis of phosphate and silicate. In this work, PLS was utilised as assistor of CIA system for simultaneous analysis of phosphate and silicate using molybdenum blue reaction without using any modification of reagents and addition of selective masking agent. The calibration ranges are 0.1–6 mgP L^?1 and 5–100 mgSi L^?1 for phosphate and silicate, respectively. By using CIA coupled with PLS for data evaluation, the analysis of two analytes was achieved within 1.5 min with only single injection. The developed system was applied to natural water samples and the system was validated with the conventional methods. By statistical paired t-test, there was no evidence of significant difference at 95% confidence level (t_stat = 2.28, t_critical = 2.31 and t_stat = 0.62, t_critical = 2.31 for phosphate and silicate, respectively). This implied that the chemometrics-assisted CIA system was successfully developed for simultaneous spectrophotometric determination of phosphate and silicate.     

A Green Deep Eutectic Solvent-Based Ultrasound-Assisted Method to Extract Astaxanthin from Shrimp Byproducts

The extraction of valuable compounds from waste products using green and inexpensive solvents is a significant strategy for sample preparation. Accordingly, in the present study, the use of deep eutectic solvents, an emerging green approach, was used to extract astaxanthin, a well-known and widely-used antioxidant, from shrimp byproducts. After evaluating different combinations of extraction conditions and deep eutectic solvents, an ultrasonication method was established and optimized by a systematic investigation of the influencing factors. A comparison of the amount of astaxanthin (102 μg g^{? 1} ) extracted using a traditional organic solvent, ethanol, showed that more astaxanthin (146 μg g^{? 1} ) was obtained using the reported eco-friendly method. The excellent properties of deep eutectic solvents highlight their advantages as promising inexpensive green solvents for the extraction and determination of a range of bioactive compounds from natural products.