RP-HPLC Column Research Articles

Retention behavior of Hg2+, MeHg+, thimerosal and phenylmercuric acetate on a C18 RP-HPLC column

Humans are exposed to potentially toxic mercuric mercury (Hg2+) and methylmercury (MeHg+) by the ingestion of food, to the bactericidal vaccine additive thimerosal (THI), and/or to the antifungal compound phenylmercuric acetate (PMA) which is used in some lens cleaning ophthalmic fluids. While numerous HPLC methods have been developed to separate Hg2+ and MeHg+ in environmental samples (e.g. food, surface waters), comparatively few have been reported for THI and PMA, in part owing to their increased hydrophobicity. We investigated the retention behavior of Hg2+, MeHg+, THI and PMA on a reversed-phase (RP) HPLC column using a flame atomic absorption spectrometer (FAAS) as a Hg-specific detector. Mobile phases comprised of 50 mM phosphate buffer (pH 7.4) with acetonitrile (ACN) concentrations of 30–50 % (v:v) produced single Hg-peaks, which eluted in the order THI, Hg2+, MeHg+ and PMA. With the 50 % ACN mobile phase, all mercurials eluted within 5 min. While the utilization of a FAAS precludes the analysis of environmental waters with the developed RP-HPLC-FAAS method, the latter is useful to probe the stability of THI and PMA in the presence of physiologically relevant concentrations of salt (100 mM in blood plasma) and l-cysteine (0.5 mM in hepatocyte cytosol), which is important as both mercurials have been recently shown to effectively inhibit the main protease of SARS-CoV-2, though the actual inhibitory Hg-species is unknown.

A new sensitive and robust method development for estimation of tolcapone and quinapril in the bulk and formulations

Tolcopone and Quinapril are anti-hypersensitive drugs and, in combined dosages, are effective in managing hypertension. The RP-HPLC method was developed using OPA and Acetonitrile in 50:50 v/v as Mobile phase and 250*4.6mm, 5µ RP-HPLC column measured with a wavelength of 240nm. The HPLC system was fixed with a flow rate of 1.0ml/min; Both drugs, TOLCO and QUINA, were successfully detected at 2.153 and 3.203 min, respectively. The validation of the method as per ICH Linearity r2 lies at 0.999, and Precision, system suitability, and selectivity RSD values are within permissible limits. The accuracy of the process was found to be 99.73 and 99.52 for TOLCO and QUINA, respectively. The LOD 0.72 and 0.3 µl/ml and LOQ 0.84 and 0.13 µl/ml. The robustness and degradation studies show that the selected method is acceptable per the Guidelines.Bottom of Form

Application of advanced environmentally benign assessment tools in determining ternary cardiovascular drug combination by RP-HPLC with analytical quality by design: Application to stability indicating method evaluation

Assessing any analytical method is becoming an important aspect for showcasing its importance and impact on the environment. The present work aimed to showcase different advanced tools available that need to be applied to analytical methods and make their developed methods essential for sustainable development. The work discussed the method development and validation of fixed combination of cardiovascular drugs, namely, cilnidipine (CIL), telmisartan (TEL), and chlorthalidone (CLD), using ICH Q14 (Recent amendment). Further application of stress studies in different conditions followed by application of recently developed assessment tools to determine the developed methodology. The developed method was optimized by using the rotatable quality by design approach, and the final optimized conditions for the separation of the mentioned drugs were performed on the Eclipse plus RP-HPLC column (C18 250 × 4.6 mm (i.d), 5 μm), with a gradient-elution mobile phase consisting of the ethanol and phosphate buffer (0.01 M, pH 3.5). Initially, the mobile phase composition was set at 40: 60. v/v (ethanol: buffer), then changed to 70: 30, v/v at 4.66 min. With a constant flow rate of 1.103 mL/min. The development method had a linearity concentration of 12.5–37.5 μg/mL for CLD, 40.0–120.0 μg/mL for TEL, and 10.0–30.0 μg/mL for CIL with a correlation coefficient (r) 0.9991, 0.9993 and 0.9991, respectively. The results of the degradation study revealed that the drugs degraded under stressed conditions, and the degradation products were separated using the same method. The degradation results indicated that the drugs degraded in stressful environments and that the degradation products have also been separated successfully using the same method. The application of greenness and whiteness evaluation tools provided comprehensive insights into the environmental impact and sustainability of the suggested methodology. The findings confirmed its favourable performance in terms of eco-friendliness and highlighted its alignment with sustainable practices. Further strengthens the credibility and relevance of the developed method in promoting a greener and more sustainable future.

Influence of Seasonal Variation on Diosgenin Content in Costus speciosus (J. Koenig) Sm. Rhizome Quantified Through Validated RP-HPLC-PDA Method

Background Costus speciosus (J. Koenig) Sm. (Syn. Cheilocostus speciosus) is an ethnic anti-diabetic plant, used for its high diosgenin content. Objectives This study aimed to evaluate the seasonal variation of diosgenin content in Costus speciosus rhizome, quantified through validated RP-HPLC method. Materials and Methods The rhizomes were collected in four different seasons, such as rainy (August), autumn (October), winter (February) and summer (May), from Lucknow, India. The HPLC method validation was done in terms of linearity, precision, repeatability, accuracy, sensitivity and robustness. Results Diosgenin was separated under isocratic elution on an RP-HPLC column (4.6 × 250 mm, 5 µm) with a mobile phase consisting of methanol and water, eluted at retention time (Rt) of 18.396 min and content was calculated with the help of standard curve. The limit of detection and limit of quantification (LOQ) was found to be 522.68 and 1583.90 ng (nanogram), respectively. The diosgenin content varies significantly in different seasons. Conclusion The diosgenin content was found higher in rainy (193.97 µg/mg) season and was concluded to be optimum season for collection of rhizomes as quality raw material. Harvesting at optimal season may fulfill the commercial demand of diosgenin and may reduce the diosgenin-intended overexploitation of the species from the wild.

Simultaneous determination of heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan glycosaminoglycan disaccharides by high-performance liquid chromatography using a reverse-phase column with adamantyl groups

Glycosaminoglycans (GAGs), which are one of the major components of proteoglycans, play a pivotal role in physiological processes such as signal transduction, cell adhesion, growth, and differentiation. Characterization of GAGs is challenging due to the tremendous structural diversity of heteropolysaccharides with numerous sulfate or carboxyl groups. In this present study, we examined the analysis of 2-aminobenzamide (2-AB) labeled GAG disaccharides by high-performance liquid chromatography (HPLC) using a reverse-phase (RP)-column with adamantyl groups. Under the analytical conditions, 17 types of 2-AB labeled GAG disaccharides derived from heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan were sequentially separated in a single analysis. The analysis time was fast with high retention time reproducibility. Moreover, the RP-HPLC column with adamantyl groups allowed the quantification of GAGs in various biological samples, such as serum, cultured cells, and culture medium.

Derivatization of sugars with N,O-dimethylhydroxylamine. Efficient RP-HPLC separation of sugar mixtures

Sugars were derivatized with N,O-dimethylhydroxylamine (DMHA) using a simple procedure. The disaccharides lactose and chitobiose and the human milk tetrasaccharides lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT) were used as examples. The β-glycosylamines were formed exclusively in good yields (80–84%). The derivatives were very well suited for RP-HPLC, giving rise to single peaks for each sugar, without the usual complications caused by mutarotation. The LNT- and LNnT-derivatives separated very well on an ordinary RP-HPLC column, despite their close structural similarity. Also, three human milk pentasaccharides (LNF I, II and III) were derivatized with DMHA. Again, good separation of these isomers was obtained. The DMHA derivatization was easily reversed. The free oligosaccharides were recovered quantitatively by mild acidic hydrolysis. To demonstrate usefulness on a preparative scale, an LNDI-rich human milk oligosaccharide fraction was derivatized, and three HPLC fractions (one major and two minor) were collected. Hydrolysis and desalting gave saccharides LNDI, LNnDII, and LNDII, the latter mixed with minor amounts of LNnDI.

Reversed Phase-HPLC Method for Low Level Quantitation of Dimethyl dibenzylidene Sorbitol

Dimethyl dibenzylidene sorbitol (DMDBS) is one of the most commonly used nucleating agent in polypropylene formulations. In present study, a simple and robust reversed phase liquid chromatography (RP-LC) based analytical method was developed for the quantitative low-level determination of DMDBS. The best separation was achieved on a RP-HPLC column TSK gel G2000 SW, 7.5mm x 30cm, Particle Size 10 and#181;m from TOSOH Bioscience. A 0.05 M NH4AC (pH = 6.6) + 10% ACN solution was used as mobile phase at a flow rate of 1.2 ml/min. UV detection was performed at 216 nm. Retention time was found to be 13.056 min. for DMDBS. The response was a linear function of concentration over the range of 2.00 to 8.00 and#181;g/ml with correlation coefficient of 0.9996 for DMDBS. The LOD and LOQ for DMDBS were found to be 1.00 and#181;g/mL and 2.00 and#181;g/mL respectively. The method is simple, rapid and robust, which is suitable for application in quality control and can also be used for the estimation of DMDBS as a leachable/ extractable from polypropylene (PP) resin and resin products like medical device containers.

In depth investigation of the retention behavior of structurally related β-blockers on RP-HPLC column: Quality by design and quantitative structure-property relationship complementary approaches for optimization and validation

The persistent introduction of new β-blockers motivates the demand for optimizing RP-HPLC well-designed analytical procedures that could be applied to this structurally related and commonly prescribed pharmacological group in order to reduce time and chemicals consumption in quality control units. Betoxolol HCl (BEX) and Carvidolol (CAR) were selected as representative examples to conduct predictive studies based on two complementary approaches, Quality by design (QBD) and Quantitative structure property relationship (QSPR). In concern QBD, a Box-Behnken design was adopted at variable chromatographic parameters to achieve the most proper conditions that might be applied for efficient analysis of the majority of group members. On the other hand, the retention time was chosen as the target property in the QSPR study that was conducted onto seven β. blockers (the two investigated drugs in addition to five other β. blockers) to find the best correlated molecular descriptors to the retention behavior. Both external and internal validation studies have comparable quality with training levels. Hence a simple selection algorithm of conventional features provides robust confirmatory predictive QBD and QSPR models. Derringer’s desirability function as as a multi-criteria approach was applied for getting the optimum chromatographic analysis conditions. Efficient analysis of BET and CAR was achieved at column temperatures of 26.00 and 27.50 °C, respectively using acetonitrile and phosphate buffer (pH 4.55) 70:30 v/v as a mobile phase with a flow rate of 1.00 mL/min, and UV detection at 220 nm. The method was validated in accordance to ICH guidelines, and had exhibited acceptable precision, accuracy, linearity, and robustness.

Furanocoumarin A: A Novel Anticancer Agent on Human Lung Cancer A549 Cells from Fructus liquidambaris.

The fruit of Fructus liquidambaris, which is recently being used for cancer treatment, has a history to be used as a traditional medicine in China for thousands of years. Ten kg of dried F. liquidambaris was obtained with 70% alcohol-water solution under reflux for three times. The condensed extract was obtained from petroleum ether, ethyl acetate and N-butyl alcohol, respectively. Ethyl acetate extract was subjected to silica gel column, Sephadex LH-20, ODS column chromatography and RP-HPLC column chromatography to yield a new compound (1). The structure was identified through intensive analysis of NMR and MS spectra. The antitumor mechanism of the furanocoumarin A on human lung cancer A549 cells was confirmed by detecting the apoptosis-related proteins. Furanocoumarin A (1), a novel furanocoumarin constituent was isolated and identified from F. Liquidambaris. The IC50 value of furanocoumarin A on A549 cell lines was 65.28±5.36µM obtained by the method of MTT. The compound could induce the apoptosis of A549 cells by inducing 21.5% early apoptosis and 32.4% late apoptosis at the concentration of 60µmol/L. Western blot analysis indicated that protein expressions of p53, caspase 3 and Bax increased in a dose-dependent manner between the concentrations from 40 to 80µM. The protein expression of Bcl-2 decreased the concentration of 60 and 80µM. The ratio of Bcl-2 to Bax was inversely proportional to the dose concentration. Furanocoumarin A could be a novel anticancer agent from herbal medicine.

Development and validation of a novel RP-HPLC method for the simultaneous quantification of ascorbic acid, gallic acid, ferulic acid, piperine, and thymol in a polyherbal formulation

A novel, accurate, precise, and linear RP-HPLC method for the simultaneous estimation of ascorbic acid, gallic acid, ferulic acid, piperine, and thymol in a polyherbal formulation was developed and validated as per ICH guidelines. A good chromatographic separation was achieved using gradient profile with the help of Shim-pack RP-HPLC column (250 mm*4.6 mm, 5 µm) and a mobile phase consisting of acetonitrile and 1% acetic acid was used. The column temperature was maintained at 28 °C throughout the run with a wavelength of 272 nm using UV-Visible detector. The retention time of ascorbic acid, gallic acid, ferulic acid, piperine, and thymol was found to be 3.9, 6.7, 21.5, 44.6, and 45.9 min respectively. The linearity of ascorbic acid, gallic acid, ferulic acid, piperine, and thymol was found to be in a range of 10-100, 5-50, 1-10, 5-50, and 2-20 µg/mL, respectively with correlation coefficient >0.99. The high recovery values (98-102%) indicate satisfactory accuracy. The % RSD values were found to be less than 2% in the precision study which reveals that the method is precise. Hence the developed method can be used for quality control and quantitative analysis of extracts and commercial containing these selected phytoconstituents.

ANALYSIS OF METABOLITES OF SYNTHETIC CANNABINOIDS JWH-018 AND JWH-073 BY CHROMATOGRAPHYMASS-SPECTROMETRIC (LC-MS/MS)METHOD IN BIOLOGICAL FLUIDS

A liquid chromatographic method was developed to resolve a comprehensive set of metabolites of JWH-018 and JWH-073. In addition to the chromatographic analysis method, an extraction method was developed to recover a broad range of synthetic cannabinoid metabolites, including carboxylic acid metabolites that are not traditionally recovered using a high pH liquid/liquid extraction. The extraction and analysis methods were used to identify and quantify the significant metabolites in urine samples. The chromatographic method detailed in this application note employed a RP-HPLC column and MS/MS detection. The quantitative range validated for all metabolites was 1 ng/mL to 500 ng/mL in urine. Based on the data, this method is suitable for quantifica- tion of metabolites of JWH-018 and JWH-073 to support broader research studies that positively identify clinically significant metabolites and their concentrations in urine.

Immunomodulatory Functions of the Human Cathelicidin LL-37 (aa 13-31)-Derived Peptides are Associated with Predicted α-Helical Propensity and Hydrophobic Index.

The anti-endotoxin activity of the cationic peptide LL-37 and its derivative IG-19 is attributed to electrostatic interaction of the peptides’ positive charge with negatively charged bacterial lipopolysaccharides (LPS), and in part to the alteration of intracellular mechanisms independent of peptide binding to LPS. We examined the immunomodulatory responses induced by IG-19 and four IG-19-derived scrambled peptides (IG-19a–d), in the presence and absence of LPS, in macrophages and peripheral blood-derived mononuclear cells. All peptides had identical net charge (+5) and amino acid composition, but different hydrophobicity and α-helical propensity. Peptide IG-19 suppressed LPS-induced cytokine/chemokine production by >90%, IG-19a and IG-19b suppressed it by 40–50%, and IG-19c and IG-19d did not suppress cytokine/chemokine production at all. In silico prediction algorithms and the peptide retention time (RT) on a C18 RP HPLC column indicated a linear association between α-helical propensity and hydrophobicity with the ability of the peptides to inhibit LPS-induced responses. Peptide RT exhibited a significant correlation (>70%) between the suppression of LPS-induced cytokine/chemokine production and peptide-induced production of the anti-inflammatory cytokine IL-1RA. These results indicate that RT on a C18 column can be used as a predictor for the immunomodulatory functions of cationic peptides. Overall, we demonstrated that the immunomodulatory functions of LL-37-derived peptides with identical positive charge and amino acid composition are directly associated with the predicted α-helical propensity and hydrophobicity of the peptides.

A new antioxidant flavonoid glycoside from flowers of Hosta plantaginea

Phytochemical investigation of the flowers of Hosta plantaginea led to isolate of one new flavonoid glycoside,plantanone C( 1) by silica gel,Sephadex LH-20,and RP-HPLC column chromatographies. Its structure was extensively determined on basis of HR-ESI-MS and NMR spectroscopic data. Compound 1 exhibited moderate antioxidant activity against DPPH radical scavenging activity,with an IC50 value of 240. 2 μmol·L~(-1).

Development and Validation of RP-HPLC Method for Estimation of Empagliflozin and its Stability Studies In vitro Simulated Gastric and Intestinal Fluids

A simple and precise high performance liquid chromatography method for determination of Empagliflozin and its stability studies invitro simulated gastric and intestinal fluids. The separation was carried out on Cosmosil C18 (250mm X 4.6, 5μ) RP-HPLC column using a 0.8ml/min as flow rate with methanol: water (80: 20 v/v) mobile phase and RT was 5.017min. Quantification was performed with a UV detector at 224nm, solubility and stability was determined individually in simulated biological fluids at various pH was investigated. The calibration curves of EMPA were linear in the range of 10–50 μg/ml(R²=0.999). The developed method was applied to pharmaceutical formulation successfully with no interfering peaks. The percentage recovery was 99.71–100.29%. It was observed that solubility of EMPA was increased in all in vitro interaction medium, freely soluble in pH 6.8 and slightly in 1.2 and 7.4 pH and the stability of EMPA was stable or less degraded in pH 6.8 for 24 hours.

Development and Validation of HPLC Method for Estimation of Pharmaceutical Drug and its Stability Studies in Simulated Biological Fluid: Comparative Study

A simple and precise high performance liquid chromatography method for determination of candesartan cilexetil in their pharmaceutical formulation was developed and validated. The separation was carried out on Cosmosil C18 (250mm X 4.6, 5μ) RP-HPLC column using an 0.8ml/min as flow rate with methanol: water (80: 20 v/v) mobile phase. Quantification was performed with a UV detector at 214nm, solubility and stability was determined individually in simulated biological fluids at various pH was investigated. The calibration curves of candesartan cilexetil were linear in the range of 10–50 μg/ml (R²=0.999). The developed method was applied to pharmaceutical formulation successfully with no interfering peaks. The percentage recovery was 99.71–100.29%. It was observed that solubility of candesartan cilexetil was increased in all invitro medium, in 1.2 and slightly 7.4 pH and the stability of candesartan cilexetil was stable in pH 1.2 or degraded in pH 7.4 for 24 hours.

Phase Ratio and Equilibrium Constant in RP-HPLC Obtained from Octanol/Water Partition Constant Through Solvophobic Theory

The experimentally known dependence in RP-HPLC of the retention factor k′ on octanol/water partition coefficient (K ow) has been examined based on solvophobic theory. The result showed that the dependence provides a means for the evaluation of phase ratio (Φ) of RP-HPLC columns, and of the equilibrium constant for a given compound and mobile phase. Using this theory, the phase ratio was evaluated for a set of seven different C18 columns (five having fully porous particles and two core–shell particles), and the equilibrium constants were calculated for four homologous series of compounds in two mobile phase systems. One mobile phase was methanol/aqueous solution of 0.1% H3PO4, and the other was acetonitrile/aqueous solution of 0.1% H3PO4. Besides providing the values for Φ for the evaluated columns, the results of the study indicated that for a specific composition of the mobile phase and for a given compound displaying only hydrophobic interactions, the equilibrium constant K(X) for different C-18 columns is basically the same. The data were further used to provide guidance in the selection of a chromatographic column for a specific separation based on K ow values and chemical structure of the analytes. The study indicated that the separation of compounds with identical polar groups (or no polar groups) and with very close values for the K ow cannot be achieved based only on hydrophobic interactions that dominate the separation on RP-type columns. Only column that displays polar interactions may provide a solution to such separations. For hydrocarbons with close K ow values, the separation cannot be achieved even on columns with some polarity. On the other hand, even compounds with equal K ow values, but with different functionalities can be separated on RP-HPLC columns without involving polar interactions. The compounds with different K ow values are expected to be easily separated on RP-HPLC columns.

Simultaneous UPLC-MS/MS determination of antiepileptic agents for dose adjustment.

Therapeutic drug monitoring (TDM) of anti-epileptic drugs (AED) is a routine application. Carbamazepine (CRB) is monitored as the parent drug while oxcarbazepine (OXC) and eslicarbazepine acetate (ESL) are monitored as their active metabolite (eslicarbazepine; MHD). We have developed a UPLC-MS/MS method for determining CRB, OXC, ESL and MHD in plasma or serum with a simplified extraction protocol. The developed method detects sildenafil (SLD), which clinically interferes with AED and is likely to be co-administered in epileptic patients suffering from sexual insufficiency (60%). MHD was prepared in-house. AED were simultaneously determined in presence of SLD using gatifloxacin as an internal standard (IS). Separation was achieved using acetonitrile, methanol and 100 mm ammonium acetate in water (32:3:65, v/v/v) on an Intersil® RP-HPLC column (250 × 4.6 mm, 5 μm). A one-step extraction was performed by simultaneous protein and phospholipids precipitation. Detection was done by tandem mass spectrometry. No relative matrix effect was observed. The method was linear (0.5-40 μg/mL for CRB, ESL and MHD and 0.05-4 μg/mL for OXC), accurate and selective. Recoveries were 64.41 ± 5.07, 45.62 ± 1.73, 61.41 ± 4.77 and 60.33 ± 1.36 for CRB, OXC, ESL and MHD, respectively. The method was successfully applied for TDM of AED.

A validated LC-MS/MS method for the estimation of glimepiride and pitavastatin in rat plasma: Application to drug interaction studies.

Glimepiride (GLI) is prescribed for the management of type-2 diabetes where as pitavastatin (PIT) for the treatment of diabetes associated dyslipidemia. Both the drugs are metabolized by CYP2C9 and have the potential of altering the enzyme through either inhibition or induction. In this respect, we present a simple, fast and validated bioanalytical LC-MS/MS method for the simultaneous estimation of GLI and PIT from rat plasma. Waters XTerra RP HPLC column (4.6×100mm, 5μm) with mobile phase consisting of acetonitrile and 10mM ammonium acetate (pH-6.0) in the ratio 85:15 (v/v) at a flow rate of 1mL/min was used for the chromatographic separation. The negative ionization mode with MRM transitions: m/z 420.17→288.13 for PIT, m/z 489.59→350.12 for GLI and m/z 380.08→316.31for celecoxib as internal standard (IS). A total run time of 3min and LLOQ was found to be 5ng/mL for both PIT and GLI. The method was applied to study the drug interaction between GLI and PIT in rat liver microsomes. In vivo rat pharmacokinetics study showed there was a 1.29-fold increase in AUC0-∞ and 1.2-fold decrease in the clearance of PIT in presence of GLI. No notable difference in the pharmacokinetic profile of GLI was observed upon the intravenous co-administration of PIT.

Liquid chromatographic enantioseparation of three beta-adrenolytics using new derivatizing reagents synthesized from (S)-ketoprofen and confirmation of configuration of diastereomers.

Diastereomers of racemic β-adrenolytic drugs [namely (RS)-propranolol, (RS)-metoprolol and (RS)-atenolol] were synthesized under microwave irradiation with (S)-ketoprofen based chiral derivatization reagents (CDRs) newly synthesized for this purpose. (S)-Ketoprofen was chosen for its high molar absorptivity (εo ~ 40,000) and its availability as a pure (S)-enantiomer. Its -COOH group was activated with N-hydroxysuccinimide and N-hydroxybenzotriazole; these were easily introduced and also acted as good leaving groups during nucleophilic substitution by the amino group of the racemic β-adrenolytics. The CDRs were characterized by UV, IR, 1 H-NMR, HRMS and CHNS. Separation of diastereomers was achieved by RP HPLC and open column chromatography. Absolute configuration of the diastereomers was established with the help of 1 HNMR supported by developing their optimized lowest energy structures using Gaussian 09 Rev. A.02 program and hybrid density functional B3LYP with 6-31G* basis set (based on density functional theory), and elution order was established. RP HPLC conditions for separation were optimized and the separation method was validated. The limit of detection values were 0.308 and 0.302 ng mL-1 . Copyright © 2016 John Wiley & Sons, Ltd.

High-performance liquid chromatography — Ultraviolet method for the determination of total specific migration of nine ultraviolet absorbers in food simulants based on 1,1,3,3-Tetramethylguanidine and organic phase anion exchange solid phase extraction to remove glyceride

The glyceride in oil food simulant usually causes serious interferences to target analytes and leads to failure of the normal function of the RP-HPLC column. In this work, a convenient HPLC-UV method for the determination of the total specific migration of nine ultraviolet (UV) absorbers in food simulants was developed based on 1,1,3,3-tetramethylguanidine (TMG) and organic phase anion exchange (OPAE) SPE to efficiently remove glyceride in olive oil simulant. In contrast to the normal ion exchange carried out in an aqueous solution or aqueous phase environment, the OPAE SPE was performed in the organic phase environments, and the time-consuming and challenging extraction of the nine UV absorbers from vegetable oil with aqueous solution could be readily omitted. The method was proved to have good linearity (r≥0.99992), precision (intra-day RSD≤3.3%), and accuracy(91.0%≤recoveries≤107%); furthermore, the lower limit of quantifications (0.05–0.2mg/kg) in five types of food simulants(10% ethanol, 3% acetic acid, 20% ethanol, 50% ethanol and olive oil) was observed. The method was found to be well suited for quantitative determination of the total specific migration of the nine UV absorbers both in aqueous and vegetable oil simulant according to Commission Regulation (EU) No. 10/2011. Migration levels of the nine UV absorbers were determined in 31 plastic samples, and UV-24, UV-531, HHBP and UV-326 were frequently detected, especially in olive oil simulant for UV-326 in PE samples. In addition, the OPAE SPE procedure was also been applied to efficiently enrich or purify seven antioxidants in olive oil simulant. Results indicate that this procedure will have more extensive applications in the enriching or purification of the extremely weak acidic compounds with phenol hydroxyl group that are relatively stable in TMG n-hexane solution and that can be barely extracted from vegetable oil.