Simultaneous determination of salbutamol sulphate, bromhexine hydrochloride and etofylline in pharmaceutical formulations with the use of four rapid derivative spectrophotometric methods

Four simple, rapid, accurate, precise, reliable and economical spectrophotometric methods have been proposed for simultaneous determination of salbutamol sulphate (SS), bromhexine hydrochloride (BH) and etofylline (ET) in pure and commercial formulations without any prior separation or purification. They were first derivative zero crossing spectrophotometry (method 1), simultaneous equation method (method 2), derivative ratio spectra zero crossing method (method 3) and double divisor ratio spectra derivative method (method 4). The ranges for SS, BH and ET were found to be 1-35 μg mL⁻¹, 4-40 μg mL⁻¹ and 5-80 μg mL⁻¹. For methods 1 and 2, the values of limit of detection (LOD) were 0.2314 μg mL⁻¹, 0.4865 μg mL⁻¹ and 0.2766 μg mL⁻¹ and the values of limit of quantitation (LOQ) were 0.7712 μg mL⁻¹, 1.6217 μg mL⁻¹ and 0.9221 μg mL⁻¹ for SS, BH and ET, respectively. For method 3, LOD values were 0.3297 μg mL⁻¹, 0.2784 μg mL⁻¹ and 0.7906 μg mL⁻¹ and LOQ values were 0.9325 μg mL⁻¹, 0.9282 μg mL⁻¹ and 2.6352 μg mL⁻¹ for SS, BH and ET, respectively. For method 4, LOD values were 0.3161 μg mL⁻¹, 0.2495 μg mL⁻¹ and 0.2064 μg mL⁻¹ and LOQ values were 0.9869 μg mL⁻¹, 0.8317 μg mL⁻¹ and 0.6879 μg mL⁻¹ for SS, BH and ET. The precision values were less then 2% R.S.D. for all four methods. The common excipients and additives did not interfere in their determinations. The results obtained by the proposed methods have been statistically compared by means of Student t-test and by the variance ratio F-test.     

Study on the interaction of nucleic acids with silver nanoparticles—Al(III) by resonance light scattering technique and its analytical application

It is found that Al(III) can further enhance the intensity of resonance light scattering (RLS) of the silver nanoparticles (AgNPs) and nucleic acids system. Based on this, a novel method of determination of nucleic acids is proposed in this paper. Under optimum conditions, there are linear relationships between the enhancing extent of RLS and the concentration of nucleic acids in the range of 1.0 × 10⁻⁹-1.0 × 10⁻⁷ g mL⁻¹, 1.0 × 10⁻⁷-2.0 × 10⁻⁶ g mL⁻¹ for fish sperm DNA (fsDNA), 1.0 × 10⁻⁹-7.0 × 10⁻⁸ g mL⁻¹ for calf thymus DNA (ctDNA) and 1.0 × 10⁻⁹-1.0 × 10⁻⁷ g mL⁻¹ for yeast RNA (yRNA). The detection limits (S/N = 3) of fsDNA, ctDNA and yRNA are 4.1 × 10⁻¹⁰ g mL⁻¹, 4.0 × 10⁻¹⁰ g mL⁻¹ and 4.5 × 10⁻¹⁰ g mL⁻¹, respectively. The studies indicate that the RLS enhancement effect should be ascribed to the formation of AgNPs-Al(III)-DNA aggregations through electrostatic attraction and adsorption bridging action of Al(III). And the sensitivity and stability of the AgNPs-fsDNA system could be enhanced by Al(III).     

Development of a validated liquid chromatographic method for the quality control of Prunellae Spica: Determination of triterpenic acids

A simple and rapid reversed-phase HPLC-UV method was developed for the determination of triterpenic acids in the crude extract of Prunellae Spica. Five triterpenic acids were extracted and isolated from P. Spica as marker compounds for use in the quality control of herbal medicines. Various solvent extraction techniques were evaluated, and the greatest efficiency was observed with sonication in 100% ethanol. Elemental compositions of the five marker compounds were determined by high-resolution mass spectroscopy. The dynamic range of the HPLC-UV method depended on the specific analyte, and acceptable quantitation was obtained between 10 and 250 μg mL⁻¹ for oleanolic acid, between 10 and 300 μg mL⁻¹ for ursolic acid, between 3 and 75 μg mL⁻¹ for 2α,3α,24-trihydroxyolean-12en-28oic acid, between 5 and 100 μg mL⁻¹ for euscaphic acid, and between 5 and 100 μg mL⁻¹ for 2α,3α-dihydroxyurs-12en-28oic acid. The method was deemed satisfactory by inter- and intra-day validation and exhibited both high accuracy and precision (relative standard deviation <9.4%). Overall limits of quantitation and detection were approximately 0.5-2.5 μg mL⁻¹ at a signal-to-noise ratio (S/N) of 3 and were about 3.0-10.0 μg mL⁻¹ at a S/N of 10. In addition, principal component analysis (PCA) was performed on the analytical data of 15 different P. Spica samples in order to classify samples collected from different regions.     

Ultrasound-assisted dispersive liquid–liquid microextraction combined with high-performance liquid chromatography-fluorescence detection for sensitive determination of biogenic amines in rice wine samples

Ultrasound-assisted dispersive liquid–liquid microextraction coupled with high-performance liquid chromatography-fluorescence detection was used for the extraction and determination of three biogenic amines including octopamine, tyramine and phenethylamine in rice wine samples. Fluorescence probe 2,6-dimethyl-4-quinolinecarboxylic acid N-hydroxysuccinimide ester was applied for derivatization of biogenic amines. Acetonitrile and 1-octanol were used as disperser solvent and extraction solvent, respectively. Extraction conditions including the type of extraction solvent, the volume of extraction solvent, ultrasonication time and centrifuging time were optimized. After extraction and centrifuging, analyte was injected rapidly into high-performance liquid chromatography and then detected with fluorescence. The calibration graph of the proposed method was linear in the range of 5–500 μg mL⁻¹ (octopamine and tyramine) and 0.025–2.5 μg mL⁻¹ (phenethylamine). The relative standard deviations were 2.4–3.2% (n = 6) and the limits of detection were in the range of 0.02–5 ng mL⁻¹. The method was applied to analyze the rice wine samples and spiked recoveries in the range of 95.42–104.56% were obtained. The results showed that ultrasound-assisted dispersive liquid–liquid microextraction was a very simple, rapid, sensitive and efficient analytical method for the determination of trace amount of biogenic amines.     

A reversed-phase high performance liquid chromatography coupled with resonance Rayleigh scattering detection for the determination of four tetracycline antibiotics

A new reversed-phase high performance liquid chromatography with resonance Rayleigh scattering detection (HPLC-RRS) was developed for simultaneous separation and determination of four tetracycline antibiotics (TCs). A good chromatographic separation among the compounds was achieved using a Synergi Fusion-RP column (150 mm × 4.6 mm; 4 μm) and a mobile phase consisting of methanol-acetonitrile-oxalic acid (5 mM) at the flow rate of 0.8 mL min⁻¹. Column temperature was 30 °C. The RRS signal was detected at λ_ex = λ_em = 370 nm. The recoveries of sample added standard ranged from 95.3% to 103.5%, and the relative standard deviation was below 2.79%. A detection limit of 2.12-5.12 μg mL⁻¹ was reached and a linear range was found between peak height and concentration in the range of 10.36-518.0 μg mL⁻¹ for oxytetracycline (OTC), 12.11-605.5 μg mL⁻¹ for tetracycline (TC), 11.79-589.5 μg mL⁻¹ for chlortetracycline (CTC) and 10.32-516.0 μg mL⁻¹ for doxycycline (DC). The linear regression coefficients were all above 0.999. The method has been applied successfully to the determination of OTC, TC, CTC, DC in pharmaceutical formulations and in honey. The method was simple, rapid and showed a better linear relation and high repeatability.     

Determination of mono- and dichloroacetic acids in betaine media by liquid chromatography

A simple and sensitive method has been developed for the analysis of residue amounts of chloroacetic acids in betaine samples based on derivatization by 1-naphthylamine (NA). The derivatized compounds are analyzed by reverse phase high performance liquid chromatography using methanol and water as mobile phase in the ratio of 32/68 (v/v) and phenyl column and PDA detection at 222 nm. The detection limits (LOD) of monochloroacetic acid (MCA) and dichloroacetic acid (DCA) are 0.1 and 0.15 μg mL⁻¹, respectively. The limits of quantification (LOQ) and the linear dynamic ranges (LDR) of MCA are found to be 1 and 1-400 μg mL⁻¹, respectively, and for DCA are found to be 3 and 3-400 μg mL⁻¹, respectively. The precision at the 5 ppm level for MCA and DCA are about 3% and 2%, (n = 5), respectively. The average recovery for MCA and DCA spiked to betaine samples are 98% and 97%, respectively.     

In situ derivatization hollow fibre liquid-phase microextraction for the determination of biogenic amines in food samples

Hollow fibre liquid-phase microextraction with in situ derivatization using dansyl chloride has been successfully developed for the high-performance liquid chromatography-ultraviolet (HPLC-UV) determination of the biogenic amines (tryptamine, putrescine, cadaverine, histamine, tyramine, spermidine) in food samples. Parameters affecting the performance of the in situ derivatization process such as type of extraction solvent, temperature, extraction time, stirring speed and salt addition were studied and optimized. Under the optimized conditions (extraction solvent, dihexyl ether; acceptor phase, 0.1 M HCl; extraction time, 30 min; extraction temperature, 26 °C; without addition of salt), enrichment factors varying from 47 to 456 were achieved. Good linearity of the analytes was obtained over a concentration range of 0.1–5 μg mL⁻¹ (with correlation coefficients of 0.9901–0.9974). The limits of detection and quantification based on a signal-to-noise ratio of 3–10, ranged from 0.0075 to 0.030 μg mL⁻¹ and 0.03 to 0.10 μg mL⁻¹, respectively. The relative standard deviations based on the peak areas for six replicate analysis of water spiked with 0.5 μg mL⁻¹ of each biogenic amine were lower than 7.5%. The method was successfully applied to shrimp sauce and tomato ketchup samples, offering an interesting alternative to liquid–liquid extraction and solid phase extraction for the analysis of biogenic amines in food samples.     

Determination of naproxen in human urine by high-performance liquid chromatography with direct electrogenerated chemiluminescence detection

A simple and sensitive liquid chromatographic method coupled with electrogenerated chemiluminescence (ECL) was described for the separation and quantification of naproxen in human urine. The method was based on the ECL of naproxen in basic NaNO₃ solution with a dual-electrode system. Factors affected the ECL emission were investigated. Under the optimal conditions, the ECL intensity has a linear relationship with the concentration of naproxen in the range of 4.0 × 10⁻⁸ g mL⁻¹ to 2.0 × 10⁻⁶ g mL⁻¹ and the detection limit was 1.6 × 10⁻⁸ g mL⁻¹ (S/N = 3). Application of the method to the analyses of naproxen in human urine proved feasible.     

Study on the interaction between oxolinic acid aggregates and protein and its analytical application

It was found that oxolinic acid (OA) at high concentration can self-assemble into nano- to micro- meter scale OA aggregates in Tris-HCl (pH 7.48) buffer solution. The nanoparticles of OA were adopted as fluorescence probes in the quantitative analysis of proteins. Under optimum conditions, the fluorescence quenching extent of nanometer scale OA aggregates was in proportion to the concentration of albumins in the range of 3.0 × 10⁻⁸ to 3.0 × 10⁻⁵ g mL⁻¹ for bovine serum albumin (BSA) and 8.0 × 10⁻⁸ to 8.0 × 10⁻⁶ g mL⁻¹ for human serum albumin (HSA). The detection limits (S/N = 3) were 3.4 × 10⁻⁹ g mL⁻¹ for BSA, and 2.6 × 10⁻⁸ g mL⁻¹ for HSA, respectively. Samples were satisfactorily determined. The interaction mechanism of the system was studied using fluorescence, UV-vis, resonance light scattering (RLS) and transmission electron microscope (TEM) technology, etc., indicating that the nonluminescent complex was formed between serum albumin molecular and OA, to disaggregate the self-association of OA, which resulted in the dominated static fluorescence quenching in the system.     

Fractionation analysis of iodine in bovine milk by preconcentration neutron activation analysis

Iodine is an essential trace element for human beings. The main source of iodine is generally food items such as fish and milk. Either the lack or the excess of iodine can cause health problems. There exists an increasing interest in the determination of total iodine as well as various species of iodine in milk. We have developed an epithermal neutron activation analysis method with a Compton suppression (ENAA-CS) counting system for the determination of ng mL⁻¹ levels of iodine. We have also employed chemical separation methods prior to ENAA-CS to measure the fraction-specific concentrations of iodine in bovine milk. We have measured the following iodine concentrations in homogenized milk (3.25%milk fat): 0.48 ± 0.02 μg mL⁻¹ of total iodine, 0.020 ± 0.003 μg mL⁻¹ of lipid-bound iodine, 0.039 ± 0.002, 0.019 ± 0.002 and 0.021 ± 0.004 μg mL⁻¹ of protein-bound iodine depending on the protein separation method and 0.45 ± 0.02 μg mL⁻¹ of inorganic species.     

Separation and determination of benzene, toluene, ethylbenzene and o-xylene compounds in water using directly suspended droplet microextraction coupled with gas chromatography-flame ionization detector

The directly suspended droplet microextraction (DSDME) technique coupled with the capillary gas chromatography-flame ionization detector (GC-FID) was used to determine BTEX compounds in aqueous samples. The effective parameters such as organic solvent, extraction time, microdroplet volume, salt effect and stirring speed were optimized. The performance of the proposed technique was evaluated for the determination of BTEX compounds in natural water samples. Under the optimal conditions the enrichment factors ranged from 142.68 to 312.13, linear range; 0.01-20 μg mL⁻¹, limits of detection; 0.8-7 ng mL⁻¹ for most analytes. Relative standard deviations for 0.2 μg mL⁻¹ of BTEX in water were in the range 1.81-2.47% (n = 5). The relative recoveries of BTEX from surface water at spiking level of 0.2 μg mL⁻¹ were in the range of 89.87-98.62%.     

Determination of indole-3-acetic acid and indole-3-butyric acid in mung bean sprouts using high performance liquid chromatography with immobilized Ru(bpy)3-KMnO4 chemiluminescence detection

A novel method for determination of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) in an extract from mung bean sprouts using high performance liquid chromatography (HPLC) with chemiluminescence (CL) detection is described. The method is based on the CL reaction of auxin (indole-3-acetic acid and indole-3-butyric acid) with acidic potassium permanganate (KMnO₄) and tris(2,2′-bipyridyl)ruthenium(II), which was immobilized on the cationic ion-exchange resin. The chromatographic separation was performed on a Nucleosil RP-C18 column (i.d.: 250 mm × 4.6 mm, particle size: 5 μm, pore size: 100) with an isocratic mobile phase consisting of methanol-water-acetic acid (45:55:1, v/v/v). At a flow rate of 1.0 mL min⁻¹, the total run time was 20 min. Under the optimal conditions, the linear ranges were 5.0 × 10⁻⁸ to 5.0 × 10⁻⁶ g mL⁻¹ and 5.0 × 10⁻⁷ to 1.0 × 10⁻⁵ g mL⁻¹ for IAA and IBA, respectively. The detection limits were 2.0 × 10⁻⁸ g mL⁻¹ and 2.0 × 10⁻⁷ g mL⁻¹ for IAA and IBA, respectively. The relative standard deviation (RSD) of intra-day were 3.1% and 2.3% (n = 11) for 2 × 10⁻⁶ g mL⁻¹ IAA and 2 × 10⁻⁶ g mL⁻¹ IBA; The relative standard deviations of inter-day precision were 6.9% and 4.9% for 2 × 10⁻⁶ g mL⁻¹ IAA and 2 × 10⁻⁶ g mL⁻¹ IBA. The proposed method had been successfully applied to the determination of auxin in mung bean sprouts.     

Determination of itopride hydrochloride by high-performance liquid chromatography with Ru(bpy)3 electrogenerated chemiluminescence detection

In this work, a stable electrogenerated chemiluminescence (ECL) detector was developed. The detector was prepared by packing cation-exchanged resin particles in a glass tube, followed by inserting Pt wires (working electrode) in this tube and sealing. The leakage of Ru(bpy)₃²⁺ can be compensated by adding a small amount of Ru(bpy)₃²⁺ into solution phase. Coupled with high-performance liquid chromatography separation, the detector has been used for determination of itopride hydrochloride in human serum. Under the optimal conditions, the ECL intensity has a linear relationship with the concentration of itopride hydrochloride in the range of 1.0 × 10⁻⁸ g mL⁻¹ to 1.0 × 10⁻⁶ g mL⁻¹ and the detection limit was 3 × 10⁻⁹ g mL⁻¹ (S/N = 3). The as-prepared ECL detector displayed good sensitivity and stability.     

Quantitative analysis of malic and citric acids in fruit juices using proton nuclear magnetic resonance spectroscopy

¹H NMR spectroscopy was applied to the quantitative determination of malic and citric acids in apple, apricot, pear, kiwi, orange, strawberry and pineapple juices. Aspartic acid was studied as a potential interference. The effect of the sample pH on the chemical shifts of signals from malic, citric and aspartic acids was examined and a value of 1.0 was selected to carry out the determination. Integration of NMR signals at 2.89-2.95 and 3.00-3.04 ppm were used for calculating the concentration of malic and citric acids, respectively. At this pH the integrated signals were not overlapped. Sodium 3-(trimethylsilyl)tetradeuteropropionate (TSP) was used as an internal reference. The obtained results applying NMR procedures to analyze the juices from different fruits were compared to those obtained using enzymatic methods and both were in close agreement. The intra- and inter-day repeatability was tested for apple juice (7.86 g l⁻¹ malic acid, 0.32 g l⁻¹ citric acid) and apricot juice (5.06 g l⁻¹ malic acid, 4.79 g l⁻¹ citric acid) obtaining coefficients of variation lower than 3.4% for intra-day measures (n = 10) and lower than 3.8% for inter-day measures (n = 20).     

A novel microchip based on indium tin oxide coated glass for contactless conductivity detection

A microfluidic chip manufactured from glass substrate and indium tin oxide (ITO) coated glass use for contactless conductivity detection was developed. The detecting electrodes were fabricated by screen-printing and chemical etching methods using an ITO-coated glass wafer. Then, the glass substrate containing separation channels was bonded with the bare side of the processed ITO-coated glass, thus producing an electrophoresis chip integrated with contactless conductivity detector. The prepared microchip displayed considerable stability and reproducibility. Sensitive response was obtained at optimal conditions (including the gap between electrodes, excitation frequency, and excitation voltage). The feasibility of this microfluidic device was examined by detection of inorganic ions, and further demonstrated by the quantification of aminopyrine and caffeine in a compound pharmaceutical. The two ingredients can be completely separated within 1 min. The detection limits were 8 μg mL⁻¹ and 3 μg mL⁻¹, respectively; with the correlation coefficient of 0.996-0.998 in the linear range from 10 μg mL⁻¹ to 800 μg mL⁻¹. The results have showed that the present method is sensitive, reliable and fast.     

Improved porous silicon microarray based prostate specific antigen immunoassay by optimized surface density of the capture antibody

Enriching the surface density of immobilized capture antibodies enhances the detection signal of antibody sandwich microarrays. In this study, we improved the detection sensitivity of our previously developed P-Si (porous silicon) antibody microarray by optimizing concentrations of the capturing antibody. We investigated immunoassays using a P-Si microarray at three different capture antibody (PSA – prostate specific antigen) concentrations, analyzing the influence of the antibody density on the assay detection sensitivity. The LOD (limit of detection) for PSA was 2.5 ng mL⁻¹, 80 pg mL⁻¹, and 800 fg mL⁻¹ when arraying the PSA antibody, H117 at the concentration 15 μg mL⁻¹, 35 μg mL⁻¹, and 154 μg mL⁻¹, respectively. We further investigated PSA spiked into human female serum in the range of 800 fg mL⁻¹ to 500 ng mL⁻¹. The microarray showed a LOD of 800 fg mL⁻¹ and a dynamic range of 800 fg mL⁻¹ to 80 ng mL⁻¹ in serum spiked samples.     

A fast derivatization procedure for gas chromatographic analysis of perfluorinated organic acids

A rapid and simple derivatization procedure has been developed for gas chromatographic determination of perfluorinated organic acids (PFCAs, C₆–C₁₂), using isobutyl chloroformate (IBCF) to convert the acids into the more volatile isobutyl esters, under catalysis by pyridine. The procedure was optimized in an acetonitrile medium and applied to GC techniques with electron-capture detection (GC-ECD) and mass spectrometry with electron-impact ionization (GC-EI-MS); for the sake of comparison, HPLC with electrospray-ionization MS (HPLC-ESI(−)-MS) was also tested. The LOD and LOQ values obtained for these three techniques were compared, and the lowest LODs were obtained with GC-ECD (0.06–1.80 μg mL⁻¹). The procedure was further optimized in an aqueous medium, obtaining the best results in a phosphate buffer (pH 2.5, 50 mmol L⁻¹), in which the LOD and LOQ values were measured for GC-ECD a GC-EI-MS. The lowest LODs were found for GC-EI-MS (0.030–0.314 μg mL⁻¹). The practical applicability was tested on Vltava river water samples.     

On-line continuous flow ultrasonic extraction coupled with high performance liquid chromatographic separation for determination of the flavonoids from root of Scutellaria baicalensis Georgi

An on-line method, based on coupling dynamic ultrasonic extraction (DUE), continuously sampling the suspension of sample and solvent, high performance liquid chromatographic separation with diode array detection, has been developed for the determination of the flavonoids, including baicalin, baicalein and wogonin, from the root of Scutellaria baicalensis Georgi. Variables influencing the DUE were evaluated by orthogonal test. The extraction yields of baicalin, baicalein and wogonin in the roots of S. baicalensis Georgi obtained from five different cultivated areas are 73.8–131.5 μg mg⁻¹ (RSD ≤ 6.24%), 6.8–15.9 μg mg⁻¹ (RSD ≤ 5.36%) and 4.4–14.3 μg mg⁻¹ (RSD ≤ 5.30%), respectively. The limits of detection for baicalin, baicalein and wogonin are 0.30, 0.37 and 0.41 μg mL⁻¹, respectively. Linearity is from 0.55 to 109 μg mL⁻¹ for baicalin, from 0.51 to 105 μg mL⁻¹ for baicalein and from 0.53 to 102 μg mL⁻¹ for wogonin. Compared with off-line continuous flow-DUE, the proposed method would be more convenient for the determination of the analytes and the rapid optimization of the extraction process. The extraction yields of flavonoids obtained by the proposed method are comparable with those obtained by dynamic microwave assisted extraction, static ultrasonic extraction and reflux extraction. The result indicated that the proposed method is suitable to determine the active components in Chinese herbal medicine.     

A rapid, acetonitrile-free, HPLC method for determination of melamine in infant formula

A simple, precise, accurate and validated, acetonitrile-free, reverse phase high performance liquid chromatography (HPLC) method is developed for the determination of melamine in dry and liquid infant formula. The separation is performed on a Kromasil C18 column (150 mm × 3.2 mm I.D., 5 μm particle size) at room temperature. The mobile phase (0.1% TFA/methanol 90:10) is pumped at a flow rate of 0.3 mL min⁻¹ with detection at 240 nm. Melamine elutes at 3.7 min. A linear response (r > 0.999) is observed for samples ranging from 1.0 to 80 μg mL⁻¹. The method provides recoveries of 97.2-101.2% in the concentration range of 5-40 μg mL⁻¹, intra- and inter-day variation in <1.0% R.S.D. The limit of detection (LOD) and limit of quantification (LOQ) values are 0.1 μg mL⁻¹ and 0.2 μg mL⁻¹, respectively.     

Gas chromatographic determination of carbonyl compounds in biological and oil samples by headspace single-drop microextraction with in-drop derivatisation

A suitable method for the gas chromatographic determination of 10 characteristic carbonyls in biological and oil samples based on the in-drop formation of hydrazones by using 2,4,6-trichlorophenylhydrazine (TCPH), has been developed. The derivatisation-extraction procedure was optimized separately for aqueous and oil samples with respect to the appropriate organic drop solvent, drop volume, in-drop TCPH concentration, sample stirring rate, temperature during single-drop microextraction (SDME), reaction time and headspace-to-sample volume ratio. The optimization showed differentiation of optimum values between the studied matrices. The limits of detection were found to range from 0.001 to 0.003 μg mL⁻¹ for the aqueous biological samples and from 0.06 to 0.20 μg mL⁻¹ for the oil samples. The limits of quantification were in the range of 0.003-0.010 μg mL⁻¹ and 0.020-0.059 μg mL⁻¹ for aqueous and oil samples, respectively. The overall relative standard deviations of the within-day repeatability and between-day reproducibility were <4.4% and <8.2% for the aqueous biological samples and <3.9% and <7.4% for the oxidized oil samples.