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Spectrophotometric Determination of Paracetamol in Urine with Tetrahydroxycalix[4]arene as a Coupling Reagent and Preconcentration with Triton X-114 Using Cloud Point Extraction

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Hayati Filik at Istanbul University-Cerrahpaşa
Hayati Filik
İzzet Şener at Kastamonu University
İzzet Şener
Sema Demirci Cekiç
Sema Demirci Cekiç
Sema Demirci Cekiç
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Emine Kilic at Ankara University
Emine Kilic
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Abstract

In the present paper, conventional spectrophotometry in conjunction with cloud point extraction-preconcentration were investigated as alternative methods for paracetamol (PCT) assay in urine samples. Cloud point extraction (CPE) was employed for the preconcentration of p-aminophenol (PAP) prior to spectrophotometric determination using the non-ionic surfactant Triton X-114 (TX-114) as an extractant. The developed methods were based on acidic hydrolysis of PCT to PAP, which reacted at room temperature with 25,26,27,28-tetrahydroxycalix[4]arene (CAL4) in the presence of an oxidant (KIO(4)) to form an blue colored product. The PAP-CAL4 blue dye formed was subsequently entrapped in the surfactant micelles of Triton X-114. Cloud point phase separation with the aid of Triton X-114 induced by addition of Na(2)SO(4) solution was performed at room temperature as an advantage over other CPE assays requiring elevated temperatures. The 580 nm-absorbance maximum of the formed product was shifted bathochromically to 590 nm with CPE. The working range of 1.5-12 microg ml(-1) achieved by conventional spectrophotometry was reduced down to 0.14-1.5 microg ml(-1) with cloud point extraction, which was lower than those of most literature flow-through assays that also suffer from nonspecific absorption in the UV region. By preconcentrating 10 ml sample solution, a detection limit as low as 40.0 ng ml(-1) was obtained after a single-step extraction, achieving a preconcentration factor of 10. The stoichiometric composition of the dye was found to be 1 : 4 (PAP : CAL4). The impact of a number of parameters such as concentrations of CAL4, KIO(4), Triton X-100 (TX-100), and TX-114, extraction temperature, time periods for incubation and centrifugation, and sample volume were investigated in detail. The determination of PAP in the presence of paracetamol in micellar systems under these conditions is limited. The established procedures were successfully adopted for the determination of PCT in urine samples. Since the drug is rapidly absorbed and excreted largely in urine and its high doses have been associated with lethal hepatic necrosis and renal failure, development of a rapid, sensitive and selective assay of PCT is of vital importance for fast urinary screening and antidote administration before applying more sophisticated, but costly and laborious hyphenated instrumental techniques of HPLC-SPE-NMR-MS.
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Pharmaceutical Society of Japan
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Pharmaceutical Society of Japan
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Pharmaceutical Society of Japan
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Pharmaceutical Society of Japan
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Pharmaceutical Society of Japan
NII-Electronic Library Service
... CPE is effectively integrated with various spectroscopic and chromatographic detection techniques, such as inductively coupled plasma-optical emission spectrometry (ICP-OES) [38], inductively coupled plasma-mass spectrometry (ICP-MS) [39,40], high-performance liquid chromatography (HPLC) with different detectors [5,6,10,12,13], liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) [8,9], α-spectrometry [41], gas chromatography (GC) [42], flame atomic absorption spectrometry (FAAS) [23,43], electrothermal atomic absorption spectrometry (ETAAS) [44], and spectrophotometry [45]. This method makes it possible to achieve a high preconcentration factor and greatly improves the selectivity of the analysis while keeping the procedure simple and environmentally friendly [3,7,8]. ...
... In a study by Filik et al. [45], a spectrophotometric method for determining paracetamol in urine was developed using tetrahydroxycalix [4]arene as a coupling reagent and preconcentration with TX-114 through cloud point extraction. The proposed CPE spectrophotometric method proved to be specific for p-aminophenol. ...
Cloud Point Extraction as an Environmentally Friendly Technique for Sample Preparation
Article
Full-text available
  • Feb 2025
Cloud point extraction is a sample preparation technique that involves using surfactants that are not harmful to the environment. It is based on micelle formation in which the extracted compound is encapsulated in the hydrophobic core of the micelles, which are the extracting agent. The most commonly used surfactants are nonionic. The others are anionic, cationic, or zwitterionic. The effectiveness of cloud point extraction might be enhanced by the addition of neutral salts, the application of proper pH, as well as acidic conditions and temperature. This sample preparation technique may be applied to extract analytes from the following matrices, such as biological and environmental samples. Cloud point extraction may be combined with various analytical techniques and detectors such as HPLC-UV, HPLC-MS, HPLC-FLD, inductively coupled plasma–optical emission spectrometry, gas chromatography, and flame atomic absorption spectrometry. When it is combined with electrothermal atomic absorption spectrometry, the limit of quantitation is low—even of the order of ng/L. The recovery of the analyte may reach the value of 100%.
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... Molecules 2020, 25, x FOR PEER REVIEW 5 of 27 effective separation of the aqueous and micellar phases. On the other hand, too high concentration of surfactant is associated with a high volume of the micelle-rich phase, and the unwanted dilution of the analyte [5,10,32]. The range of tested Triton X-114 concentrations, added to samples in multicompound determination, varied from 1.5 to 9% (w/v) [1,2,4,5,8,32], with the most frequently used concentration being 4% [32]. ...
... This was because the overly low concentration of Triton X-114 decreases recovery and prevents effective separation of the aqueous and micellar phases. On the other hand, too high concentration of surfactant is associated with a high volume of the micelle-rich phase, and the unwanted dilution of the analyte [5,10,32]. The range of tested Triton X-114 concentrations, added to samples in multi-compound determination, varied from 1.5 to 9% (w/v) [1,2,4,5,8,32], with the most frequently used concentration being 4% [32]. ...
Determination of Antidepressants in Human Plasma by Modified Cloud-Point Extraction Coupled with Mass Spectrometry
Article
Full-text available
  • Dec 2020
Cloud-point extraction (CPE) is rarely combined with liquid chromatography coupled to mass spectrometry (LC–MS) in drug determination due to the matrix effect (ME). However, we have recently shown that ME is not a limiting factor in CPE. Low extraction efficiency may be improved by salt addition, but none of the salts used in CPE are suitable for LC–MS. It is the first time that the influences of a volatile salt—ammonium acetate (AA)—on the CPE extraction efficiency and ME have been studied. Our modification of CPE included also the use of ethanol instead of acetonitrile to reduce the sample viscosity and make the method more environmentally friendly. We developed and validated CPE–LC–MS for the simultaneous determination of 21 antidepressants in plasma that can be useful for clinical and forensic toxicology. The selected parameters included Triton X-114 concentration (1.5 and 6%, w/v), concentration of AA (0, 10, 20 and 30%, w/v), and pH (3.5, 6.8 and 10.2). The addition of 10% of AA increased recovery twice. For 20 and 30% (w/v) of AA, three phases were formed that prolonged the extraction process. The developed CPE method (6% Triton X-114, 10% AA, pH 10.2) was successfully validated through LC–MS/MS simultaneous determination of 21 antidepressants in human plasma. The linearity was in the range of 10–750 ng/mL (r2 > 0.990).
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... The results of the Triton optimal concentration study showed that 1.2 mL of Triton X114 had the highest drug absorption, as shown in Fig. 7d. The findings were in line with those documented in the existing literature, as stated by the reference Filik, H. [53]. ...
Determination of candesartan and amlodipine using ion pair and cloud point methods
Article
Full-text available
  • Jan 2024
No communicable diseases are a growing concern in the field of cardiovascular disease, where the use of amlodipine and candesartan, two antihypertensive drugs, has gained significant attention as a commonly used treatment strategy. Bromothymol blue and bromocresol green were used as reagents in UV spectrophotometry to test amlodipine and candesartan. The results obtained were in the dual-ion method, assuming a linear range between 5 and 55 μg mL − 1 at pH = 11.5. The optimal dye volume is 0.7 mL. The temperature at which maximum absorption occurred was recorded at 25 • C, with corresponding recovery percentages of 98.2312 % and 102.2449 %. The range of relative standard deviation values is 0.02154-0.096745. The cloud results have a linear range extending from 1 to 55μg mL − 1. Thus, at pH = 11, the most appropriate dye volume is 1 mL. The observed recovery rates for this procedure are 98.1045 % and 100.5664 %, respectively. The relative standard deviation (RSD%) results range from 0.013205 % to 0.02427 %. The above spectral estimates were performed with a high level of accuracy, uniformity, and responsiveness. The methods described in this study have been applied in specific contexts.
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... The results of the Triton optimal concentration study showed that 1.2 mL of Triton X114 had the highest drug absorption, as shown in Fig. 7d. The findings were in line with those documented in the existing literature, as stated by the reference Filik, H. [53]. ...
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... CPE Cloud Point Extraction method which is interest with the separation method metal ions field by forming small volume of the surfactant-rich phase (colloidal system) as a result of heating non-ionic aqueous solution of surfactant to a certain temperature at which the solution become turbid ,then the colloidal solution is separated into a small volume of surfactant-rich phase and an aqueous phase (1)(2)(3)(4)(5)(6) .A list of advantages of this method than other analytical methods because of i-it green method to avoid hazardous solvents ii-its selectivity for obtaining high coefficient of preconcentration iiiits economic for capital cost iv-its rapidity for reduce the time (7)(8)(9) iiv-simplicity application for separation and purification of a variety samples of pharmaceutical products, vital and solid,and enviromental (10)(11)(12)(13)(14)(15)(16) . ...
Cloud Point Extraction, Preconcentration and Spectrophotometric Determination of Co (II) and Cu (II) using 15-Crown-5
Article
Full-text available
  • May 2021
In the current study, a cloud point extraction (CPE) with UV-vis spectrometry for detection of Cu (II) and Co (II) ions with 15-crown-5 as a chelating agent was developed to be more selective and sensitive. The CPE method is improving the analytical signal and permits the preconcentration of metals ions. After phase separation induced by change temperature to become higher more than the temperature of the cloud, the point is performed with centrifugation. The Co (II) and Cu (II) ions are determined in the enriched phase by UV-vis spectrometry. Several factors affecting the extraction efficiency were optimized, e.x. pH, the concentration of the chelating agent, non-ionic surfactant concentrations, and cloud point temperature. Under optimum conditions, a linear calibration graph in the range of 0.5-5 μg mL-1 of Co (II) in the initial solution with r 2 = 0.9994 (n=8) and 0.2-7μg of Cu (II) in the initial solution (mL-1) with r 2 = 0.9991 (n=8) were obtained. Detection limits of 0.4and 0.1μg are for Co(II) and Cu(II) along with preconcentration factors (66 and 50) for these ions, respectively. The method was used for detection of Co(II) and Cu(II) in some food samples.
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... The TX-114 was chosen as the surfactant due to its less toxicity, accessibility in purified form, high density and low cost to enable the better phase separation. Moreover, it has low cloud point temperature (about 40 °C), which allows its application for the extraction of numerous hydrophobic complexes (Filik et al. 2006). The concentration of TX-114 was studied in the range of 0.05-0.3% ...
Efficiency of different green shaking extraction methods for the preconcentration of trace quantity of mercury in artificial saliva extract of snuff products: impact on adult consumers
Article
Full-text available
  • Feb 2021
The present study is based on developing a dual-step ultrasonically modified cloud point extraction (DsUm-CPE) method for enrichment of mercury (Hg) in artificial saliva extract (ASE) of the numerous snuff products (dry and moist snuff, having green, brown and black in colors). This method relies on the complexation of Hg with ammonium O, O-diethyldithiophosphate, followed by the entrapped in nonionic surfactant (Triton X-114) prior to analysis by cold vapor atomic absorption spectrometry (CVAAS). The dispersion of hydrophobic complex in micellar phase was carried out by ultrasound energy, in addition other shaking method, vortex mixer was also applied for comparison purposes, vortex shaking method was also used. Several variables of developed methods, such as complexing agent, Triton X-114, dispersion modes (sonication/vortex shaking), equilibrium temperature, centrifugation time and concentration of back extraction reagent, have been studied. The detection and quantification limit under the optimum conditions were attained as 0.004 and 0.014 µg L−1, respectively. The accuracy for the analysis of total Hg was checked by certified samples of Virginia tobacco leaves (ICHTJ-cta-VTL-2) and both biological samples. The obtained values of all three certified samples have insignificant difference (p > 0.05) among their certified and experimental values. The resulted statistics specified that dry snuff has about fourfold higher contents of Hg as compared to moist snuff, whereas the biological samples of male subject (25 to 60 years) sniffing moist and dry snuff were also analyzed to assess the Hg exposure through their consumption. The biological samples of snuff consumers have about twofold higher concentration of Hg as compared to referents.
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... The work has aimed at developing a new spectrophotometric method with cloud point extraction (CPE) preconcentration for Salbutamol and Methyldopa by using of eosin Y as a reagent. In the CPE method, surfactant Triton X-114 was used as the extractant solution which is used for promoting phase separation [25]. ...
Application of Cloud Point Method for Spectrophotometric Determination of Salbutamol Sulphate and Methyldopa
Article
Full-text available
  • Jun 2020
Abstract A simple and efficient cloud point spectrophotometric method has been used for the determination of salbutamol sulphate and methyldopa both in pure and pharmaceutical preparations. The procedure was based on the ion association formation with eosin Y. The extraction of ion association, drown to Triton X-114 micelles, was measured spectrophotometrically. The phase separation was studied and optimized. Beer's law was rectilinear over the concentration ranges of 0.1-20 and 0.3-10 µg/mL with molar absorptivity 4x10 and 5.7x10 4 L.mol -1 cm -1 and average recovery 98.21% and 101.27% for the above drugs, respectively. The method was applied successfully for the determination of salbutamol sulphate and methyldopa in pharmaceuticals. 4
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... The work has aimed at developing a new spectrophotometric method with cloud point extraction (CPE) preconcentration for Salbutamol and Methyldopa by using of eosin Y as a reagent. In the CPE method, surfactant Triton X-114 was used as the extractant solution which is used for promoting phase separation [25]. ...
Application of Cloud Point Method for Spectrophotometric Determination of Salbutamol Sulphate and Methyldopa
Article
Full-text available
  • Jun 2020
A simple and efficient cloud point spectrophotometric method has been used for the determination of salbutamol sulphate and methyldopa both in pure and pharmaceutical preparations. The procedure was based on the ion association formation with eosin Y. The extraction of ion association, drown to Triton X-114 micelles, was measured spectrophotometrically. The phase separation was studied and optimized. Beer's law was rectilinear over the concentration ranges of 0.1-20 and 0.3-10 µg/mL with molar absorptivity 4x104 and 5.7x104 L.mol-1 cm-1 and average recovery 98.21% and 101.27% for the above drugs, respectively. The method was applied successfully for the determination of salbutamol sulphate and methyldopa in pharmaceuticals.
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Facile synthesis of cotton flower like Ni–Co/Ni–Co–O–P as bifunctional active material for alkaline overall water splitting and acetaminophen sensing
Article
  • Aug 2022
  • INT J HYDROGEN ENERG
The development of electrode materials with simple preparation, favorable price, excellent electrocatalytic activity, and stability are some of the most important issues in the field of electrochemistry. Herein, we prepared Ni–Co/Ni–Co–O–P cotton flower like on a copper sheet (CS) by a convenient, efficient, and scalable electrodeposition method. The Ni–Co/Ni–Co–O–P was employed as effective binder free electrode material in two different applications such as electrocatalytic water splitting and acetaminophen (APAP) sensor. Remarkably, the Ni–Co/Ni–Co–O–[email protected] exhibits low overpotentials of 310 and 90 mV at 10 mA cm⁻² for oxygen and hydrogen evolution reactions in alkaline media, respectively. Besides, the Ni–Co/Ni–Co–O–[email protected] || Ni–Co/Ni–Co–O–[email protected] couple needs a low cell voltage of 1.62 V to achieve a current density of 10 mA cm⁻², and its potential change is negligible after 20 h of continuous operation. Furthermore, Ni–Co/Ni–Co–O–P displays good electrochemical sensing performance toward APAP with a high sensitivity of 803.74 μA mM⁻¹cm⁻², low limit of detection of 0.16 μM, a wide linear range of 0.05 mM–3 mM, and a fast response time of 3.3 s. This work proposes a simple approach for synthesis of Ni–Co/Ni–Co–O–P as an efficient electrode material for water splitting and APAP sensing.
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Determination of Rhodamine B by UV–Vis spectrophotometry in cosmetics after microextraction by using heat-induced homogeneous liquid–liquid extraction method
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An environmentally friendly microextraction method which includes determination of trace levels of rhodamine B in cosmetic and detergent samples by using UV–vis spectrophotometric determination after enrichment with has been established heat-induced homogeneous liquid–liquid microextraction method. As extraction solvent, cyclohexylamine has been used, and the rhodamine B concentration in extraction phase was determined at 550 nm by using UV–vis spectrophotometry. The major parameters influencing in the method including pH, volume of cyclohexylamine, amount of NaCl, sample volume have been assessed. The influences of the matrix components were also investigated. The limit of detection, the limit of quantitation and linear range were found as 0.014, 0.047 and 0.047–4.79 μg mL−1, respectively. The relative standard deviation was 1.03%. The extraction procedure was applied to determination of rhodamine B in the cosmetic and detergent samples.
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Determination of acetaminophen and phenacetin in plasma by high pressure liquid chromatography
Article
Full-text available
  • Jul 1977
We describe a sensitive and precise high-pressure liquid chromatographic method in which acetoacetanilide is used as the internal standard to simultaneously determine acetaminophen and phenacetin in plasma. Therapeutic as well as toxic concentrations can be determined on as little as 0.1 ml of plasma. Sample preparation is rapid and chromatography is complete in 5 min. Quantitation is accurate at 0.5 mg/liter concentration for both drugs. Day-to-day precision within 5% is attainable. Of 36 other drugs tested, only theophylline interfered, with the determination of acetaminophen.
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Screening for Paracetamol Poisoning
Article
  • Jan 1973
The commonly used tests for the detection of paracetamol or its metabolites in cases of self-poisoning are either non-specific or time consuming. The need for screening all cases of self-poisoning for paracetamol ingestion is emphasised and a test is recommended which combines the advantages of increased specificity with those of a short procedure.
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A Validated High-Performance Liquid Chromatographic Method for the Determination Of Paracetamol and Its Major Metabolites in Urine
Article
  • Sep 1995
  • J Liq Chrom
A reversed-phase high-performance liquid chromatographic (RP-HPLC) method employing UV-detection (248 nm), 5 μm Resolve C18 as stationary phase (in a 8 × 100 mm Waters Radial-Pak cartridge) and 0.1 M potassium phosphate monobasic/methanol/glacial acetic acid (95:4:1, v/v/v) as mobile phase was developed for the rapid and simultaneous determination of paracetamol and its major metabolites (its glucuronide and sulphate conjugates) in urine. The method requires only minimal sample preparation and chromatographic run time is only 6 min. For determination of paracetamol, the precision (inter-assay RSD ranged from 1.60–0.66% between 5 and 100 μg mL) and limits of detection (2 ng mL) were satisfactory, as were these parameters for determination of the major metabolites. In the course of studies on paracetamol bioavailability and metabolism, over 1500 samples have been assayed using this method. Herein we report its use for monitoring of the levels of paracetamol and its major metabolites in the urine of fifteen normal healthy volunteers given a single oral dose of 500 mg.
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Usefulness of the Evaporative Light Scattering Detector for Direct Screening of Biological Fluids
Article
  • May 2001
  • ANAL CHIM ACTA
This paper reports a fast, simple procedure for the direct screening of biological fluids by using a flow configuration coupled on-line to an evaporative light scattering detector with no chromatographic separation. The potential of this coupling was exemplified for paracetamol. The configuration adopted permits selective retention of paracetamol and its metabolites from dilute urine or serum samples on a LiChrolut®EN sorbent column located in the loop of a high pressure injection valve. Retained paracetamol is eluted with acetonitrile and driven directly to the evaporative light scattering detector. Some analytical features are: detection limit 0.3μgml−1, quantitation interval 1–100μgml−1, repeatability, as RSD, 1.0%. The reliability of the method was studied at three concentration levels of paracetamol spiked to urine or serum blanks (1, 2 and 4 times the detection limit) yielding 0% of false positives and negatives, thanks to the high precision of the method. Finally a comprehensive study of 50 real urine samples (paracetamol free) obtained from different individuals (healthy, drug addicts and patients from a hospital) provided 4% of false positives and 0% false negatives for the threshold limit established.
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Catalytic oxidation and flow detection of acetaminophen at a dicyanobis(1,10-phenanthroline)iron(II)-modified electrode
Article
  • Jun 1994
  • ANAL CHIM ACTA
Dicyanobis(1,10-phenanthroline)iron(II)-modified glassy carbon electrodes were shown to exhibit an electrocatalytic response for the oxidation of acetaminophen with a decrease of 100 mV in the potential required. It can also inhibit the oxidation of ascorbic acid and uric acid. When used for amperometric monitoring of acetaminophen in a flowing stream, the coated electrode may permit detection at lower potentials compared with the naked surface. The method gives a linear range from 0.05 to 100 μg ml−1 and a detection limit of 0.02 μg ml−1 (signal-to-noise ratio = 2). The application of the method to the determination of acetaminophen in urine by LC is briefly described.
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High‐Pressure Liquid Chromatographic Determination of Acetaminophen in Biological Fluids
Article
  • Jul 1976
  • J PHARM SCI-US
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Liquid-chromatographic determination of acetaminophen in serum
Article
  • Apr 1979
We describe a sensitive and precise "high-pressure" liquid-chromatographic method for determining acetaminophen in serum. The 2-acetaminophenol and 3-acetaminophenol structural isomers of acetaminophen are used as internal standards. The method, which involves solvent extraction and adsorption chromatography on silica, provides excellent sensitivity, accuracy, and selectivity. The standard curve is linear over the range of acetaminophen concentrations of 0.5 to 300 mg/L, which makes the method useful for both pharmacokinetic studies and overdose monitoring. Analytical recovery is 97% for acetaminophen concentrations ranging from 5 to 300 mg/L. Many commonly used drugs were tested and found not to interfere. The procedure has been successfully adapted as a microscale method requiring only 50 microL of sample. The microscale method is particularly useful for pediatric and neonatal patients for whom sample size is a major concern.
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Emergency determination of acetaminophen
Article
  • Oct 1978
I describe a senstive colorimetric micromethod in which a pH 11.0 carbonate buffer is used to induce acetaminophen to react with Folin-Ciocalteau reagent at room temperature. These reagents and a simple solvent extraction favor formation of a stable indophenol dye with acetaminophen, giving this procedure a good degree of specificity. Results correlate well with those by liquid-chromatographic procedures, and day-to-day precision is less than 8%.
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