Determination Of Retinol Research Articles

Cyclic Voltammetry of Retinol in Surfactant Media and Its Application for the Analysis of Real Samples

AbstractRetinol is irreversibly oxidized at 790 mV in water 0.1 M LiClO4 that corresponds to formation of retinal. Effect of surfactant on voltammetric characteristics of retinol oxidation has been evaluated. The calibration graph was linear in the range 29.4–980 µM of retinol with detection limit of 15 µM in presence of 1.1×10−4 M of sodium dodecyl sulfate. Application of surfactant media leads to decrease of detection limit, enlargement of analytical range for retinol determination and permits analyze retinol in water media. Simple, express and reliable voltammetric approach for determination of retinol in real samples using surfactant media have been developed.

Vitamin A Is Systemically Bioavailable After Intratracheal Administration With Surfactant in an Animal Model of Newborn Respiratory Distress

Chronic lung disease (CLD) is a major cause of long-term morbidity in extremely LBW infants with respiratory distress syndrome. Parenteral vitamin A administration decreases the risk of CLD. We tested the hypothesis that intratracheal vitamin A administration with surfactant is systemically bioavailable without interfering with the functional properties of exogenous surfactant. Newborn piglets were ventilated with 100% FiO2 and sequential saline lavage induced respiratory distress syndrome. During lung injury induction, ventilator changes were allowed, but none were made following treatment allocation. Animals were assigned by chance in a blinded control trial to three groups: I=control; II=surfactant; III=surfactant+vitamin A. Hemodynamics, lung mechanics, and blood gases were measured following instrumentation, pre- and posttreatment for 4 h, at which time the liver was sampled for retinol determination. All parameters improved in animals receiving surfactant. A significant interaction existed between time and group for PaO2 and alveolar-arterial oxygen difference (A-aDO2). Hepatic levels of retinol were higher (p<0.001) in animals receiving retinyl acetate. Intratracheal administration of surfactant+vitamin A did not alter the beneficial effects of surfactant on lung compliance and gas exchange. Intratracheal Vitamin A was associated with rapid hepatic uptake. Further studies are warranted.

Determination of retinol, retinyl palmitate, and retinoic acid in consumer cosmetic products

Retinol and retinyl palmitate are frequently used in cosmetic products. A simple, rapid, and sensitive reversed-phase high-performance liquid chromatography (HPLC) method with ultraviolet (UV) detection was developed for the quantitation of retinol, retinyl palmitate, and retinoic acid in cosmetic preparations. The analytes were extracted from a cosmetic/Celite mixture using a solvent system composed of equal amounts of hexane, isopropanol, and ethyl acetate, and the extract was injected directly into an HPLC chromatograph with a C18 column and UV detector set at 330 nm. Chromatographic separation was achieved by gradient elution with a mobile phase, starting with aqueous ammonium acetate buffer/methanol that was gradually changed to methanol/dichloromethane. The average recoveries of retinol, retinyl palmitate, and retinoic acid from spiked cosmetic products were 95% or higher. In a survey of twenty-nine consumer cosmetic skin care products labeled to contain retinoids, most products were found to contain either retinol or retinyl palmitate at concentrations up to 2.2% (w/w), while a few products contained both ingredients. A number of products also contained cis isomers of retinol that could be quantitatively distinguished from the all-trans compound. The method can be used to quantitate several retinoids and their isomers in cosmetic products. The method will be useful for obtaining information needed to estimate levels of exposure to retinoids from cosmetic products.

Suitability of ultra-high performance liquid chromatography for the determination of fat-soluble nutritional status (vitamins A, E, D, and individual carotenoids)

Our aim was to assess the suitability of ultra-high performance liquid chromatography (UHPLC) for the simultaneous determination of biomarkers of vitamins A (retinol, retinyl esters), E (alpha- and gamma-tocopherol), D (25-OH-vitamin D), and the major carotenoids in human serum to be used in clinical practice. UHPLC analysis was performed on HSS T3 column (2.1 x 100 mm; 1.8 microm) using gradient elution and UV-VIS detection. The system allows the simultaneous determination of retinol, retinyl palmitate, 25-OH-vitamin D, alpha- and gamma-tocopherol, lutein plus zeaxanthin, alpha-carotene, beta-carotene, alpha- and beta-cryptoxanthin and lycopene. The method showed a good linearity over the physiological range with an adequate accuracy in samples from quality control programs. Suitability of the method in clinical practice was tested by analyzing samples (n = 286) from patients. In conclusion, UHPLC constitutes a reliable approach for nutrient/biomarker profiling allowing the rapid, simultaneous and low-cost determination of vitamins A, E, and D (including vitamers and ester forms) and the major carotenoids in clinical practice.

Miniaturisation of solid phase extraction method for determination of retinol, alpha- and gamma-tocopherol in human serum using new technologies

The aim of this study was to develop rapid and simple solid phase extraction (SPE) and HPLC methods for simultaneous determination of retinol, gamma- and alpha-tocopherol in human serum using a special auto sampler with micro titration plates. Separation of vitamins was performed at ambient temperature using monolithic column on a HPLC containing rack changer for micro titration plates. As the mobile phase methanol with flow rate 2.5 mL min−1 was used. The injection volume was 20 µL. Retinol was detected at 325 nm, gamma- and alpha-tocopherol were carried out at 295 nm, respectively. The total time of analysis was 1.8 minutes. Extraction method was developed using Spe-ed 96 C18, 100 mg/2 mL micro titration plates and SPE vacuum manifold. The consumption of the sample was 50 µL. Time of the analysis for 96 samples on one micro titration plate was 1.5 hour. In order to validate the developed method, precision, accuracy, linearity, detection and quantitation limits were evaluated. This method is suitable for rapid automated large-batch analysis of retinol, alpha- and gamma-tocopherol in small sample volumes of human serum.

Simultaneous determination of retinol, retinyl palmitate and β-carotene in rat serum treated with 7,12-dimethylbenz[a]anthracene and Hypericum Perforatum L. by high-performance liquid chromatography with diode-array detection

Abstract A new and simple high-performance liquid chromatography method was developed and validated for the simultaneous determination of retinol, retinyl palmitate and β-carotene in rat serum treated with Hypericum Perforatum L. and 7,12-dimethylbenz[a]anthracene. Furthermore, vitamin C was determined spectrophotometrically. High-performance liquid chromatography analysis was performed utilizing an Inertsil ODS3 reversed phase column with methanol-acetonitrile-tetrahydrofuran (65:30:5, v/v/v) as mobile phase, at a flow rate of 1.5 mL min−1 and 40°C. Diode-array detection was conducted at 325 and 450 nm for retinol and retinyl palmitate, and β-carotene, respectively with a running time of 26 min. The high-performance liquid chromatography assay and extraction procedure proposed are simple, rapid, sensitive and accurate. This method was then applied to determine the amounts of retinol, retinyl palmitate and β-carotene in rat serum. Results of this study demonstrated that at 60th day in the 7,12-dimethylbenz[a]anthracene-treated group there was a significant decrease (pa] anthracene + Hypericum Perforatum L. treated group compared to the control group..

Solid phase extraction with electrospun nanofibers for determination of retinol and α-tocopherol in plasma

A novel packed-fiber solid phase extraction procedure based on electrospun nanofibers for simultaneous determination of vitamins A (retinol) and E (α-tocopherol) in human plasma has been developed. Parameters affecting extraction efficiency were investigated in detail. The limit of detection is 0.01 μg mL−1 for retinol, and 0.3 μg mL−1 for α-tocopherol. The linear range is from 0.05 to 2.0 μg mL−1 for retinol, and from 0.5 to 30 μg mL−1 for α-tocopherol. The precision (RSD) is 90%. The method was applied to analysis of retinol and α-tocopherol in human plasma with satisfactory results.

Plasma Levels of Vitamins A and E, Coenzyme Q10, and Anti-ox-LDL Antibody Titer in Children Treated with an Elimination Diet Due to Food Hypersensitivity

Has elimination diet applied in children with food hypersensitivity in infancy any effect on plasma levels of anti-oxidative vitamins and antibodies to oxidized low-density lipoprotein (anti-ox-LDL antibody) titer in these children at their pre-school age?" The study involved 92 children (3 to 7 years of age) with food hypersensitivity treated in their infancy and early childhood with soy formula or casein hydrolysate, as a milk substitute for at least 12 months. Control group comprised 62 children, who had never been treated with an elimination diet. The status of the anti-oxidative system was evaluated by determination of retinol, alpha-tocopherol, and coenzyme Q10 plasma levels by high-performance liquid chromatography (HPLC). The titer of antibodies to oxidized LDL lipoproteins was specified by immunoenzymatic assay. On the basis of the RESULTS, the following CONCLUSIONS have been reached: 1. It was shown that alpha-tocopherol and retinol levels in pre-school children who had received dietary treatment in their infancy, were higher than in the control group. No deficiencies in anti-oxidative vitamins within the control group were found. 2. A type of milk-substitute formula applied in the elimination diet had no effect on the status of the anti-oxidative system in the children examined.

Evaluation of uncertainty of measurement from method validation data: An application to the simultaneous determination of retinol and α-tocopherol in human serum by HPLC

An isocratic RP-HPLC method for the determination of retinol and α-tocopherol in serum, with fluorescence and UV/VIS detection, respectively, was developed and validated according to international guidelines. Detection (retinol 0.015 mg/L, α-tocopherol 0.29 mg/L) and quantification (retinol 0.05 mg/L, α-tocopherol 0.95 mg/L) limits were determined. Repeatability was <3.3% and <2.9% and intermediate precision was <4.6% and <3.2%, for retinol and α-tocopherol, respectively. Certified reference materials were utilised to assess bias and guarantee traceability to SI units. Expanded uncertainties (retinol 8.9%; α-tocopherol 7.9%), estimated according to the EURACHEM/CITAC guide from method validation data, satisfied fit-for-purpose requirements based on biological variability.

Flow-Injection Determination of Retinol and Tocopherol in Pharmaceuticals with Acidic Potassium Permanganate Chemiluminescence

A simple and rapid flow-injection method is reported for the determination of retinol, its derivatives and alpha-tocopherol by its enhancement effect on a potassium permanganate-formaldehyde chemiluminescence system in an acidic medium. Linear calibration graphs were obtained between 5.0 x 10(-8) to 5.0 x 10(-6) mol L(-1) for all-trans-retinol, 5.0 x 10.8 - 2.5 x 10(-6) mol L(-1) for all-trans-retinal, 5.0 x 10(-7) - 1.0 x 10.5 mol L(-1) for retinoic acid, 1.0 x 10(-7) - 5.0 x 10(-6) mol L(-1) for retinyl acetate and retinyl palmitate and 1.0 x 10(-8) - 5.0 x 10(-6) for alpha-tocopherol, with relative standard deviations (RSD; n = 4) in the range of 0.8 - 2.8%. The limits of detection (S/N = 3) were in the range of 5.0 x 10(-9) - 1.0 x 10(-7) mol L(-1) with a sample throughput of 100 h(-1). The influence of common ions, excipients in pharmaceutical preparations and related organic compounds were studied on the determination of retinol and alpha-tocopherol individually. The method was applied to determine retinol and alpha-tocopherol in pharmaceutical preparations, and the results obtained were in reasonable agreement with the amount labelled.

Isocratic rapid liquid chromatographic method for simultaneous determination of carotenoids, retinol, and tocopherols in human serum

An improved isocratic and rapid HPLC method was developed for the measurement of carotenoids, retinol and tocopherols in human serum. Vitamins were extracted with hexane. Mobile phase consisted of a mixture acetonitrile:methylene chloride:methanol with 20 mM ammonium acetate. This method used a small bead size (3 μm) Spherisorb ODS2 column with titane frits. Diode array and fluorescence detectors were used respectively for the detection of carotenoids and retinol/tocopherols. Chromatographic separation was complete in 13 min for β-cryptoxanthin, cis–trans-lycopene, α-carotene, β-carotene, cis-β-carotene, retinol, δ-tocopherol, γ-tocopherol and α-tocopherol. Echinenone and tocol were employed as internal standards for diode array and fluorescence detection. Accuracy was validated using standard reference material (SRM) 968C. Intra-assay and inter-assay precision were respectively 0.2–7.3% and 3.6–12.6%. Sensitivity was verified using the ICH recommendations and the limit of detection (LOD) obtained was sufficient for routine clinical application.

Lycopene for Advanced Hormone Refractory Prostate Cancer: A Prospective, Open Phase II Pilot Study

We investigated the influence of lycopene on the clinical and laboratory course in men with hormone refractory prostate cancer. To our knowledge this study represents the first time that subjective assessments of the course of therapy have been recorded. We performed a prospective, open phase II pilot study, in which patients with progressive hormone refractory prostate cancer were included. Lycopene supplementation (15 mg) was given daily for 6 months. Followup laboratory tests and clinical examinations were done monthly. Changes to analgesic use and quality of life (European Organisation for Research and Treatment of Cancer QLQ-C30) were measured. The study end point was a significant change in serum prostate specific antigen, clinical progression or the end of the 6-month observation period. A total of 18 patients 64 to 85 years old (median age 73) were enrolled in the study during a 20-month period, of whom 17 could be analyzed. Five of the 17 patients (29%) withdrew from the study prematurely, including 4 of 5 because of prostate specific antigen progression and/or tumor associated complications, and 1 due to an allergic reaction to lycopene. Median prostate specific antigen doubled in 6 months from 42.7 ng/ml (range 13.8 to 521.6) in 17 patients to 96.4 ng/ml (range 13.5 to 1,240) in 12. Stable prostate specific antigen was observed in 5 of 17 patients (29%). None of the patients had a greater than 50% decrease in prostate specific antigen. Patients experienced a slight deterioration in mean health status at the end of the study compared to the outset. However, two-thirds of the patients experienced an improved or unchanged situation regardless of the clinical and biochemical course. No clinically relevant benefits were shown for patients with advanced stages of the disease.

The role of retinol in the etiology and outcome of suppurative otitis media

Vitamin A deficiency is still a serious public health problem affecting an estimated 127 million preschool children. The resulting immunological dysfunctions lead to increased risks of respiratory tract infections, diarrhoeal diseases and blindness, among others. The aim of this study is to determine the significance of the role of serum retinol in the etiology of acute suppurative otitis media (ASOM) and its chronicity (CSOM). In a prospective follow-up of patients with ASOM for 6-9 months, serum retinol determination was done using the high performance liquid chromatography. Participants comprised 358 ASOM and 52 control subjects. Six-month follow-up was achieved in 264 subjects (74%); of these, there was persistence of otorrhoea (CSOM) in 116, while 148 had resolved ASOM. Of the 264 subjects, 146 were males and 118 were females, between the ages of 6 months and 9 years and a mean of 7 years (SD = 2.32), whereas the control subjects comprised 29 males and 22 females, between the ages of 6 months and 11 years and a mean of 7.8 years (SD = 3.6). The range of serum retinol in the ASOM subjects was 1.63-2.64 microg/L, mean of 1.53 microg/L, median value of 2.61 microg/L and (SD = 0.16). Among control subjects, the range was 2.5-2.8 microg/L, mean of 2.58 microg/L and median value of 2.61 microg/L (SD = 0.14) (Table 1). The range of serum retinol in the resolved ASOM subjects was 1.61-2.63 microg/L, mean of 2.07 microg/L and median value of 2.09 microg/L (SD = 0.16) while the CSOM subjects ranged between 0.8-2.86 microg/L, mean of 1.58 microg/L and median value of 1.28 microg/L, (SD = 0.48) (Table 2). Univariate analysis using unpaired t test to compare the mean serum retinol revealed significant difference between ASOM and control (P = 0.0000) and between resolved ASOM and CSOM (P = 0.0000). In conclusion, hyporetinolaemia was a significant etiological factor in the etiology ASOM and CSOM, suggesting retinol supplementation as one strategy in control of SOM.

Simultaneous determination of retinol and α-tocopherol in polymeric diets for enteral nutrition

Existing methods for simultaneous measurements of retinol and α-tocopherol in enteral formulas require large sample and solvent volumes and are time-consuming and costly. We have developed a simple, sensitive, cost-effective method for the determination of these vitamins in polymeric diets that can easily be applied to standard quality control of large numbers of samples. Our analytical procedure comprises deproteinization with pure ethanol, saponification with a 3.6 M KOH solution in a sonicator for 30 min at 65 °C under a nitrogen atmosphere, solubilization of samples in phosphate buffer and extraction with hexane. Vitamins are separated by reversed-phase HPLC and quantified by dual-wavelength spectrophotometry. The method gives satisfactory results, with recovery rates of 106.3 ± 1.5% for retinol and 102.3 ± 1.5% for α-tocopherol and RSDs ranging between 1.2 and 4.8% for precision. This method is suitable for the quality control of enteral formulas.

Simultaneous determination of benzophenone‐3, retinol and retinyl acetate in pig ear skin layers by high‐performance liquid chromatography

A new and simple HPLC method was developed and validated for the simultaneous determination of benzophenone-3, retinol and retinyl acetate in pig ear skin layers and percutaneous penetration samples after in vitro permeation experiments. HPLC analysis was performed utilizing a NovaPak C18 column with acetonitrile-water-acetic acid as mobile phase. UV detection was at 325 nm and the run time was 25 min. The detector response was found to be accurate, precise and linear across the analytical range. Analyte extraction from skin layers was done with methanol from the stratum corneum and epidermis, and with acetone from the dermis. Recovery was in all cases better than 90%. The HPLC assay and extraction procedure proposed are simple, rapid, sensitive and accurate. The method was then applied for the determination of benzophenone-3, retinol and retinyl acetate in pig ear skin layers after topical application.

Simultaneous measurement of retinol, α-tocopherol and six carotenoids in human plasma by using an isocratic reversed-phase HPLC method

A simple and sensitive isocratic reversed-phase high-performance liquid chromatography (HPLC) method for simultaneous determination of retinol, α-tocopherol and six carotenoids in human plasma was described. Sample preparation of the earlier published method was further developed by addition of ultrapure water, which enabled aqueous layer to freeze facilitating phase separation without pipetting thus also improving precision of the method. Developed method appeared to be less laborious and time consuming compared to the traditional extraction methods, which require removal of organic layer by pipetting. The recoveries (absolute and relative) were between 80% and 103%. The intra-assay CVs were 1.1–4.0% (normal level) and 3.3–9.0% (low level). Inter-assay CVs were 5.3–8.8%. Reference method for all these analytes was not available, but a comparison with another published method was carried out. The results of the comparison matched satisfactorily. The method is used routinely in our laboratory in a large population-based study.

Improvement in Retinol Analysis by Fluorescence and Solid Phase Extraction (SPE) in Micellar Medium

In this paper, the determination of retinol, the structure with the most activity as vitamin A, was carried out in an aqueous micellar medium with a low quantity of a short-chain alcohol. The analytical technique used in this work was fluorescence, which gave us very much information qualitative and quantitative. The sensitivity of the method is higher than that obtained in other media; the detection limit is 0.03 mg L(-1) and retinol was stable in solution for at least 5 days. The use of solid phase extraction (SPE) for organic samples, allowed us to change the organic matrix by a mixture CTAB 5%/n-butanol 10%/water 85% w/w/w with recoveries in retinol spiked samples close to 100%. In addition, the combination of SPE and fluorescence is a good preconcentration technique, sensitive and fast for the identification and determination of retinol, simultaneously.

Effect of low vitamin A diets with high-moisture or dry corn on marbling and adipose tissue fatty acid composition of beef steers

Angus-cross steers (n = 165; 295 +/- 16 kg of BW) were used evaluate the effect of low vitamin A diets with high-moisture corn (HMC) or dry corn (DC) on marbling and fatty acid composition. Steers were allotted to 24 pens (7 steers/pen), such that each pen had the same average initial BW. Treatments were randomly allotted to the pens. The experiment had a completely randomized design, with a 2 x 2 factorial arrangement of treatments: low vitamin A (Lo, no supplemental vitamin A) and HMC (LoHMC); LoDC; high vitamin A (Hi, supplemented with 2,200 IU of vitamin A/kg of DM) and HMC (HiHMC); and HiDC. Diets contained 76% corn, 10% corn silage, 11% protein supplement, and 3% soybean oil (DM basis). Samples of feed ingredients were collected for carotenoid analysis. Blood samples were collected for serum retinol determination. Steers were slaughtered after 145 d on feed. Carcass characteristics and LM composition were determined. Samples from the s.c. fat depot were analyzed for fatty acid composition. High-moisture corn had a greater vitamin A content, based on its carotenoid content, than DC (614 vs. 366 IU/kg of DM, P < 0.01). No vitamin A x corn type interactions were detected for feedlot performance, carcass characteristics, or serum, s.c. fat, or liver retinol concentration. Average daily gain, DMI, and G:F were not affected by vitamin A (P > 0.05). Marbling score and USDA quality grade were greater (P < 0.05) in Lo vs. Hi steers. Hot carcass weight, backfat, and yield grade were not affected by the treatments (P > 0.05). Vitamin A and corn type did not affect LM composition (DM, ash, CP, or ether-extractable fat, P > 0.05). Vitamin A supplementation increased (P < 0.06) serum retinol on d 112 and 145 and increased (P < 0.01) liver retinol at slaughter (Lo = 38.7 vs. Hi = 102.9 mug/g). The s.c. fat retinol concentrations were less (P < 0.01) for Lo (0.8 mug/g) than for Hi (1.4 mug/g) at slaughter. Cell diameter of adipocytes in the i.m. depot was not affected by dietary vitamin A (P > 0.05). A vitamin A x corn type interaction was observed (P < 0.05) for the s.c. fat cellularity. Feeding HMC increased the number of cells per square millimeter when Lo diets were fed (LoHMC = 128 vs. LoDC = 100 cells/mm(2), P < 0.05), but not when Hi diets were fed (HiHMC = 109 vs. HiDC = 111 cells/mm(2), P > 0.05). The CLA content of adipose tissue was not affected by the treatments. Regardless of the corn type used, feeding low vitamin A diets for 145 d to Angus-cross steers increased marbling and quality grade without affecting yield grade, animal health, or performance.

Comparison of a novel ultra-performance liquid chromatographic method for determination of retinol and α-tocopherol in human serum with conventional HPLC using monolithic and particulate columns

Retinol and alpha-tocopherol are biologically active compounds often monitored in blood samples because of their evident importance in human metabolism. In this study a novel ultra-performance liquid chromatographic (UPLC) method used for determination of both vitamins in human serum has been compared with conventional HPLC with particulate and monolithic C(18) columns. In UPLC a sub-two-micron particle-hybrid C(18) stationary phase was used for separation, in contrast with a five-micron-particle packed column and a monolithic column with a highly porous structure. Methanol, at flow rates of 0.48, 1.5, and 2.5 mL min(-1), respectively, was used as mobile phase for isocratic elution of the compounds in the three methods. Detection was performed at 325 nm and 290 nm, the absorption maxima of retinol and alpha-tocopherol, respectively. Analysis time, sensitivity, mobile-phase consumption, validation data, and cost were critically compared for these different chromatographic systems. Although cost and mobile-phase consumption seem to make UPLC the method of choice, use of the monolithic column resulted in almost the same separation and performance with a slightly shorter analysis time. These methods are alternatives and, in routine laboratory practice, more economical means of analysis of large numbers of biological samples than use of a traditional particulate column.

Development and validation of a liquid chromatography method for the simultaneous determination of α‐tocopherol, retinol and retinyl esters in human serum using a monolithic column for the monitoring of anticancer therapy side effects

Among other side effects, administration of anticancer agents is accompanied by manifestations of gastrointestinal toxicity and disturbances of antioxidant balance. The monitoring of these toxic effects in clinical practice is impeded by a dearth of reliable laboratory methods. Therefore, a simple and rapid reversed-phase high-performance liquid chromatography procedure for selective and sensitive determination of retinol, a-tocopherol, and retinyl esters (retinyl-palmitate and retinyl-stearate) in blood serum has been developed and presented in this study. A Series 200 LC HPLC instrument from Perkin Elmer (Norwalk, USA) with diode-array detector (DAD) was used for the analysis. Separations of retinol, alpha-tocopherol, retinyl-palmitate, and retinyl-stearate were performed using a Chromolith Performance RP-18e, 100 x 4.6 mm monolithic column from Merck (Darmstadt, Germany). Gradient elution was used at a flow rate of 3 mL/min; the mobile phase was methanol-water (95:5, v/v) for 0-2.1 min and methanol-2-propanol (60:40, v/v) for 2.1-4.9 min. The total time of analysis was 6 min. The injection volume was 20 microL and the analysis was performed at ambient temperature. Detection of retinol, alpha-tocopherol, and retinyl esters was carried out at 325, 295, and 330 nm, respectively. For practical assessment of the method, the vitamin A absorption test was performed on seven healthy controls as well as on six patients with non-small cell lung carcinoma or head and neck carcinoma previously treated by chemotherapy and/or radiotherapy, six patients with rectal carcinoma before chemoradiotherapy, four patients with gastrointestinal stromal tumor (GIST) before treatment with imatinib, and a breast cancer patient with chemotherapy-induced diarrhea. Present data demonstrate the feasibility of large scale HPLC determination of vitamin E, vitamin A, and retinyl esters in human serum using a silica monolithic column, and this method may represent a valuable aid in the laboratory monitoring of the toxicity of anticancer therapy.