Reproducibility Relative Standard Deviation Research Articles

Determination of Gliadin as a Measure of Gluten in Food by R5 Sandwich ELISA RIDASCREEN® Gliadin Matrix Extension: Collaborative Study 2012.01.

BackgroundAccording to Codex Alimentarius, food products containing less than 20 mg/kg gluten can be labeled as “gluten-free.” Since 2002, the R5 antibody method allowed determination of gluten levels and led to a huge improvement of products available to celiac disease (CD) patients.MethodThe R5-containing test kit RIDASCREEN® Gliadin in combination with the cocktail solution was endorsed as Codex Type 1 Method in 2006 based on a collaborative study with corn-based bread, rice-based dough, wheat starches, rice, and corn flour. In 2012, the method was approved as First Action Official MethodSM2012.01 with an “in foods” claim. For Final Action in 2016, the matrix claim was reduced to rice- and corn-based matrixes.ObjectiveTherefore, R-Biopharm decided to start a collaborative study to demonstrate the wide applicability of Official Method 2012.01 for the quantitative analysis of gliadin in soy, starches, pseudo cereals, legumes, spices, juice, nut nougat crème, cream cheese, pesto, meat, vegetarian meat alternative, cookies, dessert, cake, fish, bread, candies, and potatoes. Materials for incurring were the MoniQA wheat flour and the PWG gliadin preparation. ResultsGliadin levels ranged from 3.4 up to 27.4 mg gliadin per kg. The results of the collaborative study with 14 participating laboratories showed recoveries ranging from 80 to 130%. Relative reproducibility standard deviations for contaminated samples were between 9.8 and 27.7%.ConclusionsThe collaborative study results confirmed that the method is accurate and suitable to measure gliadin in important gluten-free food matrixes.HighlightsThe title and applicability statement of Official Method 2012.01 were changed as proposed.

Fiber Nanoarchitectonics for Pre-Treatments in Facile Detection of Short-Chain Fatty Acids in Waste Water and Faecal Samples.

Short-chain fatty acids (SCFAs) are among the active metabolites in biological process both in the intestinal tract and the bioconversion of organic wastes, which has resulted in various human diseases and environmental problems. In order to accurately detect SCFAs, we introduced a novel extraction sorbent. Electrospun polyacrylonitrile (PAN) nanofiber membrane was synthesized, then poly (3, 4-Ethylenedioxythiophene) (PEDOT) was deposited onto the surface of electrospun PAN nanofibers by in situ polymerization. The morphology of the composite PAN/PEDOT nanofiber was characterized by scanning electronic microscopy (SEM) and FTIR spectrum. PAN/PEDOT was used to isolate and concentrate the SCFAs in waste water and fecal samples before gas chromatography mass spectrometry (GC-MS) analysis. The analytical method was evaluated systematically, and low limits of detection (LODs) of 0.34–0.87 μg/L and good linearity (R2 ≥ 0.9953) were obtained. The method was applied successfully for the determination of SCFAs in waste water and fecal samples, with good recovery (87.5–104.6%) and satisfactory reproducibility (relative standard deviation: 6.5–14.1%). The results indicated that the proposed method can be used as a potential approach for the determination of SCFAs with high sensitivity in waste water and biological samples.

Real-Time PCR Assay for the Detection and Quantification of Roe Deer to Detect Food Adulteration-Interlaboratory Validation Involving Laboratories in Austria, Germany, and Switzerland.

Game meat products are particularly prone to be adulterated by replacing game meat with cheaper meat species. Recently, we have presented a real-time polymerase chain reaction (PCR) assay for the identification and quantification of roe deer in food. Quantification of the roe deer content in % (w/w) was achieved relatively by subjecting the DNA isolates to a reference real-time PCR assay in addition to the real-time PCR assay for roe deer. Aiming at harmonizing analytical methods for food authentication across EU Member States, the real-time PCR assay for roe deer has been tested in an interlaboratory ring trial including 14 laboratories from Austria, Germany, and Switzerland. Participating laboratories obtained aliquots of DNA isolates from a meat mixture containing 24.8% (w/w) roe deer in pork, roe deer meat, and 12 meat samples whose roe deer content was not disclosed. Performance characteristics included amplification efficiency, level of detection (LOD95%), repeatability, reproducibility, and accuracy of quantitative results. With a relative reproducibility standard deviation ranging from 13.35 to 25.08% (after outlier removal) and recoveries ranging from 84.4 to 114.3%, the real-time PCR assay was found to be applicable for the detection and quantification of roe deer in raw meat samples to detect food adulteration.

Development, optimization, and validation of a method for detection of cartap, thiocyclam, thiosultap-monosodium, and thiosultap-disodium residues in plant foods by GC-ECD

A method was developed for the simultaneous determination of four nereistoxin-related pesticides, viz. cartap, thiocyclam, thiosultap-monosodium, and thiosultap-disodium, in 20 plant foods. The samples were extracted using a hydrochloric acid solution containing cysteine hydrochloride, derivatized to nereistoxin under alkaline conditions, and analyzed by gas chromatography with electron capture detector. The average recoveries of the method were 72–108%, with relative standard deviations (RSDs) of 0.3–14.7% (n = 1200, p < 0.05). The intermediate precision and reproducibility experiments using established methods were also carried out. All the results passed the Cochrane and Grubbs tests (n = 2400, p < 0.05). The RSDs of intermediate precision and RSDs of reproducibility among laboratories were in the ranges 1.7–10.9% and 2.4–15.3% (n = 2400, p < 0.05), respectively, indicating that the accuracy and precision of the method are satisfactory. This method can be used to detect nereistoxin-related pesticides in plant foods.

Determination of camostat and its metabolites in human plasma – Preservation of samples and quantification by a validated UHPLC-MS/MS method

ObjectivesCamostat mesilate is a drug that is being repurposed for new applications such as that against COVID-19 and prostate cancer. This induces a need for the development of an analytical method for the quantification of camostat and its metabolites in plasma samples. Camostat is, however, very unstable in whole blood and plasma due to its two ester bonds. The molecule is readily hydrolysed by esterases to 4-(4-guanidinobenzoyloxy)phenylacetic acid (GBPA) and further to 4-guanidinobenzoic acid (GBA). For reliable quantification of camostat, a technique is required that can instantly inhibit esterases when blood samples are collected. Design and methodsAn ultra-high-performance liquid chromatography-tandem mass spectrometry method (UHPLC-ESI-MS/MS) using stable isotopically labelled analogues as internal standards was developed and validated. Different esterase inhibitors were tested for their ability to stop the hydrolysis of camostat ester bonds. ResultsBoth diisopropylfluorophosphate (DFP) and paraoxon were discovered as efficient inhibitors of camostat metabolism at 10 mM concentrations. No significant changes in camostat and GBPA concentrations were observed in fluoride-citrate-DFP/paraoxon-preserved plasma after 24 h of storage at room temperature or 4 months of storage at −20 °C and −80 °C. The lower limits of quantification were 0.1 ng/mL for camostat and GBPA and 0.2 ng/mL for GBA. The mean true extraction recoveries were greater than 90%. The relative intra-laboratory reproducibility standard deviations were at a maximum of 8% at concentrations of 1–800 ng/mL. The trueness expressed as the relative bias of the test results was within ±3% at concentrations of 1–800 ng/mL. ConclusionsA methodology was developed that preserves camostat and GBPA in plasma samples and provides accurate and sensitive quantification of camostat, GBPA and GBA by UHPLC-MS/MS.

Determination of silanol group content on the surface of fumed silica by chemical reaction-headspace gas chromatography

建立了一种基于化学反应-顶空气相色谱测定气相二氧化硅表面硅羟基含量的新方法。实验取气相二氧化硅放入顶空瓶中于105 ℃烘箱中加热2 h去除水分,将甲苯稀释的格氏试剂注入密闭的顶空瓶中,格氏试剂与气相二氧化硅表面硅羟基快速反应产生甲烷(CH4),甲烷量与气相二氧化硅表面硅羟基含量成正比。经过气相色谱-氢火焰离子化检测器测定甲烷,通过外标法定量,根据化学反应方程式计算出样品中羟基含量。同时对反应溶液用量与反应时间等条件进行优化,确定2.0 mL反应溶液,反应15 min为最优的前处理条件。结果表明,硅羟基含量与气相色谱信号值之间存在良好的线性相关性,相关系数为0.9990,相对标准偏差小于3%,本方法的检出限为0.30 mg/g,定量限为1.00 mg/g,开展了4家实验室对5个不同比表面积的样品测试,数据结果的重复性限(r)小于2.5%,再现性限(R)小于6.5%。该方法结合自动化技术,顶空反应操作简单,样品量和试剂用量少,准确性高,重复性好,优于酸碱滴定法,适用于快速检测气相二氧化硅表面硅羟基的含量,解决了硅羟基利用传统方法难以准确测定的难题。该方法的建立对我国二氧化硅产业硅羟基检测标准的制定和产业技术优化,均具有重要的理论和现实意义。

An LC-MS/MS Method for the Determination of Antibiotic Residues in Distillers Grains: Collaborative Study.

Antibiotics are used in ethanol production to discourage the growth of bacteria that would lower the yield of the product. Any antibiotic residues remaining in distillers grain (DG) co-product could lead to antimicrobial resistance, which is a public health concern. The U.S. Food and Drug Administration (FDA) Center for Veterinary Medicine (CVM) previously developed an LC-MS/MS analytical method to detect residues of erythromycin A, penicillin G, virginiamycin M1, and virginiamycin S1 in DG to enable regulatory decision making. The objective of this study was to ensure the method's robustness by carrying out a multi-laboratory validation of the method. Test portions were extracted with a mixture of acetonitrile and buffer. The extract was cleaned by solid phase extraction. The concentrated eluant was reconstituted and analyzed by LC-MS/MS. Eight laboratories participated in the study. Average accuracies for the combined three matrixes for all four compounds at all fortification levels ranged from 83->109% with repeatability relative standard deviation (RSDr; within laboratory) ≤17% and reproducibility relative standard deviation (RSDR; between laboratory) ≤21%. The Horwitz ration (HorRat) values ranged 0.4-1.0 indicating that method reproducibility is acceptable. An interlaboratory study was successfully conducted to evaluate an LC-MS/MS method for the determination of the drugs of interest in DG. The results demonstrate that the method is fit for purpose to determine the drugs in DG and could serve as a regulatory method capable of being used for compliance actions for DG containing these antibiotic contaminants. The method was posted to the FDA/Foods Program Compendium of Analytical Laboratory Methods.

Au\u2013Ag assembled on silica nanoprobes for visual semiquantitative detection of prostate-specific antigen

BackgroundBlood prostate-specific antigen (PSA) levels are widely used as diagnostic biomarkers for prostate cancer. Lateral-flow immunoassay (LFIA)-based PSA detection can overcome the limitations associated with other methods. LFIAbased PSA detection in clinical samples enables prognosis and early diagnosis owing to the use of high-performance signal reporters.ResultsHere, a semiquantitative LFIA platform for PSA detection in blood was developed using Au–Ag nanoparticles (NPs) assembled on silica NPs (SiO2@Au–Ag NPs) that served as signal reporters. Synthesized SiO2@Au–Ag NPs exhibited a high absorbance at a wide wavelength range (400–800 nm), with a high scattering on nitrocellulose membrane test strips. In LFIA, the color intensity of the test line on the test strip differed depending on the PSA concentration (0.30–10.00 ng/mL), and bands for the test line on the test strip could be used as a standard. When clinical samples were assessed using this LFIA, a visual test line with particular color intensity observed on the test strip enabled the early diagnosis and prognosis of patients with prostate cancer based on PSA detection. In addition, the relative standard deviation of reproducibility was 1.41%, indicating high reproducibility, and the signal reporter showed good stability for 10 days.ConclusionThese characteristics of the signal reporter demonstrated the reliability of the LFIA platform for PSA detection, suggesting potential applications in clinical sample analysis.

A validated UHPLC-MS/MS method for rapid determination of senicapoc in plasma samples

The clinically tested KCa3.1 channel blocker, senicapoc, has been proven to have excellent pharmacological properties and prior clinical trials found it to be safe for use in patients with sickle cell anaemia. Currently, several preclinical projects are aiming to repurpose senicapoc for other indications, but well-described analytical methods in the literature are lacking. Our aim was to develop a sensitive, rapid and accurate ultra-high-performance liquid chromatography-tandem mass spectrometry method using pneumatically assisted electrospray ionisation (UHPLC-ESI-MS/MS) suitable for the determination of senicapoc in plasma samples. Unfortunately, direct analysis of senicapoc in crude acetonitrile extracts of human plasma samples by UHPLC-ESI-MS/MS was subjected to significant and variable ion suppression from coeluting phospholipids (PLs). The interferences were mainly caused by the presence of phosphatidylcholine and phosphatidylethanolamine classes of PLs, including their lyso-products. However, the PLs were easily removed from crude extracts by filtration through a sorbent with Lewis acid properties which decreased the total ion suppression effect to approximately 5%. Based on this technique, a simple high-throughput UHPLC-MS/MS method was developed and validated for the determination of senicapoc in 100-μL plasma samples. The lower limit of quantification was 0.1 ng/mL. The mean true extraction recovery was close to 100 %. The relative intra-laboratory reproducibility standard deviations of the measured concentrations were 8% and 4% at concentrations of 0.1 ng/mL and 250 ng/mL, respectively. The trueness expressed as the relative bias of the test results was within ± 2% at concentrations of 1 ng/mL or higher.

Magnetic molecularly imprinting polymers, reduced graphene oxide, and zeolitic imidazolate frameworks modified electrochemical sensor for the selective and sensitive detection of catechin.

A glassy carbon electrode (GCE) was modified with magnetic molecularly imprinted polymers (mMIPs) using catechin as a template, reduced graphene oxide (rGO), and zeolitic imidazolate frameworks-8 (ZIF-8) for the sensitive detection of catechin (mMIPs/rGO-ZIF-8/GCE). The prepared rGO, ZIF-8, and mMIPs exhibited typical structures and properties determined by various characterizations. The mMIPs showed good selectivity for catechin among several structural analogs. The mMIPs/rGO-ZIF-8/GCE showed a higher maximum peak current for catechin than that of a single component modified GCE. After the optimization of the material ratio, coating amounts, pH, and scan rate, the mMIPs/rGO-ZIF-8/GCE exhibited good selectivity, good linearity, and a low detection limit (LOD) for catechin. The linear range was 0.01nmol/L-10μmol/L and the LOD was 0.003nmol/L (S/N = 3). The relative standard deviations for reproducibility and stability tests (n = 6) were 5.2% and 6.1%, respectively. A recovery between 99.1 and 101.3% was obtained in the detection of catechin in spiked samples. Based on these findings, the proposed mMIPs/rGO-ZIF-8/GCE could be developed further, and future research could be conducted on alternate fabrication strategies and methods to create more portable and practical electrochemical sensors. Graphical Abstract.

Validation of a multi-analyte HPLC method for the determination of carotenoids used as feed additives in fish and poultry feed: results of an interlaboratory study

ABSTRACT The performance characteristics of a multi-analyte method for the determination of all 10 carotenoids authorised as feed additives within the EU were assessed via an interlaboratory study. The analytical method is based on reversed phase high performance liquid chromatography (RP-HPLC) coupled to an optical detector set at 410 nm. The analysis is particularly challenging due to the presence of various stereoisomers of each carotenoid, and the use of these compounds via natural or synthetic formulations, requiring a special sample preparation. EU regulations specifying the conditions of use set legal limits for these substances in compound feedingstuffs ranging from 6 mg kg−1 to 138 mg kg−1, depending on the individual carotenoid and the target animal for which the feed is supplemented with this carotenoid. The purpose of the multi-analyte method validated in this paper is to facilitate the monitoring of carotenoids at relevant levels when used as feed additives in compound feedingstuffs and pre-mixtures. The interlaboratory study delivered precision data for 43 different analyte/mass fraction/matrix combinations, covering a mass fraction range of the target analytes from 2.6 mg kg−1 to 3861 mg kg−1. The relative standard deviations for repeatability (RSDr) varied from 2.2 to 16.2 % with a mean value of 6 %, while the relative standard deviations for reproducibility (RSDR) varied from 6.8 to 39 % with a mean value of 21 %. Given the broad scope of the method covering 10 carotenoids added to compound feedingstuffs and pre-mixtures via different formulations, this multi-analyte method is considered fit for the intended purpose.

Determination of Zearalenone and Trichothecenes, Including Deoxynivalenol and Its Acetylated Derivatives, Nivalenol, T-2 and HT-2 Toxins, in Wheat and Wheat Products by LC-MS/MS: A Collaborative Study.

An analytical method for the simultaneous determination of trichothecenes—namely, nivalenol (NIV), deoxynivalenol (DON) and its acetylated derivatives (3- and 15-acetyl-DON), T-2 and HT-2 toxins—and zearalenone (ZEN) in wheat, wheat flour, and wheat crackers was validated through a collaborative study involving 15 participants from 10 countries. The validation study, performed within the M/520 standardization mandate of the European Commission, was carried out according to the IUPAC (International Union of Pure and Applied Chemistry) International Harmonized Protocol. The method was based on mycotoxin extraction from the homogenized sample material with a mixture of acetonitrile-water followed by purification and concentration on a solid phase extraction column. High-performance liquid chromatography coupled with tandem mass spectrometry was used for mycotoxin detection, using isotopically labelled mycotoxins as internal standards. The tested contamination ranges were from 27.7 to 378 μg/kg for NIV, from 234 to 2420 μg/kg for DON, from 18.5 to 137 μg/kg for 3-acetyl-DON, from 11.4 to 142 μg/kg for 15-acetyl-DON, from 2.1 to 37.6 μg/kg for T-2 toxin, from 6.6 to 134 μg/kg for HT-2 toxin, and from 31.6 to 230 μg/kg for ZEN. Recoveries were in the range 71–97% with the lowest values for NIV, the most polar mycotoxin. The relative standard deviation for repeatability (RSDr) was in the range of 2.2–34%, while the relative standard deviation for reproducibility (RSDR) was between 6.4% and 45%. The HorRat values ranged from 0.4 to 2.0. The results of the collaborative study showed that the candidate method is fit for the purpose of enforcing the legislative limits of the major Fusarium toxins in wheat and wheat-based products.

One-Step Rapid Synthesis of Al-Based Ag Dendrites for Highly Active and Cost Effective SERS Plasmonic Substrates

Highly active Al-based Ag dendrites SERS plasmonic substrates have been rapidly synthesized by the one-step galvanic displacement reaction without the use of any surfactants and templates. The as-prepared SERS substrates were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and Raman spectroscopy. XRD measurements confirmed the metallic nature of the formed Ag dendrites. None of the organic additives were used in the synthesis process, which ensures the substrates surfaces are completely clean and avoiding the introduction of organic contaminants. The innovative rough bionic substrates yield a final silver dendritic structure that offers large specific surface area and high-density hotspots. Using malachite green as a model target, the Al-based Ag dendrites SERS substrates exhibited acceptable reproducibility (relative standard deviation of 23.8%) and high enhancement capacities (pushed the detection limit down to 10 pM). Importantly, these Ag dendrites could potentially be employed as highly active and cost effective flexible SERS sensors for label-free ultrasensitive detection of biomolecules.

Particle size analysis of pristine food-grade titanium dioxide and E 171 in confectionery products: Interlaboratory testing of a single-particle inductively coupled plasma mass spectrometry screening method and confirmation with transmission electron microscopy

Titanium dioxide is a white colourant authorised as food additive E 171 in the EU, where it is used in a range of alimentary products. As these materials may contain a fraction of particulates with sizes below 100 nm and current EU regulation requires specific labelling of food ingredient to indicate the presence of engineered nanomaterials there is now a need for standardised and validated methods to appropriately size and quantify (nano)particles in food matrices.A single-particle inductively coupled plasma mass spectrometry (spICP-MS) screening method for the determination of the size distribution and concentration of titanium dioxide particles in sugar-coated confectionery and pristine food-grade titanium dioxide was developed. Special emphasis was placed on the sample preparation procedure, crucial to reproducibly disperse the particles before analysis. The transferability of this method was tested in an interlaboratory comparison study among seven experienced European food control and food research laboratories equipped with various ICP-MS instruments and using different software packages. The assessed measurands included the particle mean diameter, the most frequent diameter, the percentage of particles (in number) with a diameter below 100 nm, the particles' number concentration and a number of cumulative particle size distribution parameters (D0, D10, D50, D99.5, D99.8 and D100). The evaluated method's performance characteristics were, the within-laboratory precision, expressed as the relative repeatability standard deviation (RSDr), and the between-laboratory precision, expressed as the relative reproducibility standard deviation (RSDR). Transmission electron microscopy (TEM) was used as a confirmatory technique and served as the basis for bias estimation.The optimisation of the sample preparation step showed that when this protocol was applied to the relatively simple sample food matrices used in this study, bath sonication turned out to be sufficient to reach the highest, achievable degree of dispersed constituent particles. For the pristine material, probe sonication was required. Repeatability and reproducibility were below 10% and 25% respectively for most measurands except for the lower (D0) and the upper (D100) bound of the particle size distribution and the particle number concentration. The broader distribution of the lower and the upper bounds could be attributed to instrument-specific settings/setups (e.g. the timing parameters, the transport efficiency, type of mass-spectrometer) and software-specific data treatment algorithms. Differences in the upper bound were identified as being due to the non-harmonised application of the upper counting limit. Reporting D99.5 or D99.8 instead of the effectively largest particle diameter (D100) excluded isolated large particles and considerably improved the reproducibility. The particle number-concentration was found to be influenced by small differences in the sample preparation procedure. The comparison of these results with those obtained using electron microscopy showed that the mean and median particle diameter was, in all cases, higher when using spICP-MS. The main reason for this was the higher size detection limit for spICP-MS plus the fact that some of the analysed particles remained agglomerated/aggregated after sonication.Single particle ICP-MS is a powerful screening technique, which in many cases provides sufficient evidence to confirm the need to label a food product as containing (engineered) titanium dioxide nanomaterial according to the current EU regulatory requirements. The overall positive outcome of the method performance evaluation and the current lack of alternative standardised procedures, would indicate this method as being a promising candidate for a full validation study.

Collaborative study on the determination of 7-aminoflunitrazepam in urine by GC-MS

A previously published gas chromatography-mass spectrometry (GC-MS) method for the analysis of 7-aminoflunitrazepam (7-amino-FM2) in urine was subjected to an inter-laboratory collaborative study. The method was rigorously defined in terms of performance requirements, yet allowed some degrees of flexibility to individual analysts. The collaborative study followed the guidelines provided by the Association of Official Analytical Chemists International (AOAC International). Each of nine drug-abuse urine testing laboratories in Taiwan participated in the study to analyze 3 samples in duplicate. The repeatability relative standard deviation (RSDr) and the reproducibility relative standard deviation (RSDR) of the measurements of 7-amino-FM2 in urine were determined to be less than 9.91% and 14.84%, respectively. The method showed acceptable reproducibility, as evidenced by HORRAT values of 0.6 to 0.7. The method was thus proposed to be used by the testing laboratories in Taiwan.

Determination of pentazocine in urine by gas chromatography-mass spectrometry

A sensitive and specific gas chromatography-mass spectrometry (GC-MS) method using selected ion monitoring (SIM) has been developed for quantification of pentazocine in urine. This method uses solid-solid extraction (SPE) and N, O-bis (trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane derivatization, followed by GC-MS analysis using dextromethorphan as the internal standard. The method was linear between 125 and 1500 ng/mL (r2 = 0.997). The limit of quantification and limit of detection were 125 and 62.5 ng/mL, respectively. The intraday accuracy and precision were -6.3∼4.7% and 3.6∼11.2%, respectively. The interday accuracy and precision were -7.5∼7.0% and 6.2∼10.0%, respectively. These data showed that the GC-MS method was suitable for the determination of pentazocine in urine. This method was further applied to an interlaboratory study. The collaborative study was given by the Association of Official Analytical Chemists International (AOAC International). Nine drug abuse urine testing laboratories in Taiwan participated to analyze 3 samples in duplicate. The repeatability relative standard deviation (RSDr) and reproducibility relative standard deviation (RSDR) values were 3.0∼5.2% and 6.8∼10.3%, respectively. The method showed acceptable reproducibility, as evidenced by HORRAT values of 1.1 to 1.5. The method recommended for use by the drug abuse urine testing laboratories in Taiwan.

Collaborative study on the determination of phencyclidine in urine by gas chromatography-mass spectrometry

A gas chromatography-mass spectrometry method for the analysis of phencyclidine (PCP) in urine was subjected to an inter-laboratory study. The collaborative study followed the guidelines provided by the Association of Official Analytical Chemists International. Ten laboratories participated, analyzing 3 samples of PCP-spiked urine as blind duplicate. The repeatability relative standard deviation (RSDr) and the reproducibility relative standard deviation (RSDR) were between 2.1%-3.6% and 4.2%-7.3%, respectively. HORRAT values for the reproducibility showed 0.4-0.7, indicating acceptable precision between laboratories. The method was thus proposed to be used by the drug-abuse urine testing laboratories in Taiwan.

Simultaneous Determination of Sesquiterpene Lactones in Ixeris chinensis by HPLC

A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of six sesquiterpene lactones: 3β-[3-(4-hydroxyphenyl)acetyl-β-D-glucopyranosyloxy]-8β-hydroxy-4(15),10(14)11(13)-guaiatrien-12,6-olide (AR1) 3β-[3-(4-hydroxyphenyl)acetyl-β-D-glucopyranosyloxy]-4(15),10(14)11(13)-guaiatrien-12,6-olide (AR2), 3β-(β-D-glucopyranosyloxy)-8β-(4-hydroxyphenyl)acetoxy-4(15),10(14)11(13)-guaiatrien-12,6-olide (AR3), 3β-(β-D-glucopyranosyloxy)-8β-(2-hydroxy-3-methylbutanoyloxy)acetoxy-4(15),10(14)11(13)-guaiatrien-12,6-olide (AR4), 3β-(β-D-glucopyranosyloxy)-4(15),10(14)11(13)-guaiatrien-12,6-olide (AR5), and 3β-hydroxy-4(15),10(14)11(13)- guaiatrien-12,6-olide (AR6) in the whole plant of Ixeris chinensis. The separation by gradient elution was performed on Hypersil ODS-2 column (250 mm × 4.6 mm, 5 µm) at 30°C with acetonitrile and water as the mobile phase, and monitored by absorbance at 238 nm. The parameters of linearity, precision, accuracy, and specificity of the method were evaluated. The recovery of the method is 95.85-98.19%, and linearity (r ＞ 0.9993) was obtained for all sesquiterpene lactones. A high degree of specificity as well as repeatability and reproducibility (relative standard deviation values less than 2.0%) were also achieved. This assay was successfully applied to the determination of six sesquiterpene lactones in ten samples. The results indicated that the developed assay method was rapid, accurate, reliable and could be readily utilized as a quantitative analysis method for I. chinensis.

Determination of fluoroquinolones in milk, honey and water samples by salting out-assisted dispersive liquid-liquid microextraction based on deep eutectic solvent combined with MECC.

A simple and sensitive salting out-assisted dispersive liquid-liquid microextraction method using deep eutectic solvent combined with back extraction and micellar electrokinetic capillary chromatography (SO-DLLME-DES-BE-MECC) was developed for the determination of fluoroquinolones in milk, honey and water samples. Several parameters affecting the extraction efficiency including DES volume, vortex time, centrifugation time, salt type and amount, sample pH and volume, etc. were investigated. Good linearity were obtained for fluoroquinolones in a range of 0.020-3.200μgmL-1 and 0.030-4.800μgmL-1 with LODs less than 0.010μgmL-1. The recoveries were in the ranges of 95.0-104.9%, 90.1-110.2% and 87.8-114.1% for water, honey and milk samples, respectively. The relative standard deviations for reproducibility were all below 7.6%. Under the optimized conditions, the enrichment factors for analytes were achieved in the range from 531 to 858 folds. The presented method was successfully applied for the determination of fluoroquinolones in milk, honey and water samples.

Clay-Protein Nanocomposite Based Electrochemical Sensor for the Determination of Ascorbic Acid

This paper describes sensitive ascorbic acid (AA) determination on montmorillonite clay and silk protein sericin nanocomposite (MMT-Ser NC) modified pencil graphite electrode (PGE) for the first time. Nanocomposite structures were synthesized by desolvation technique and cross-linked with glutaraldehyde (GA). The synthesized MMT-Ser NC was evaluated by various methods. As a result of the characterization studies, it was determined that the sericin was successfully converged with the montmorillonite structure, and that the MMT-Ser NCs were homogeneous and uniform, with the size of 150 nm and zeta potentials of approximately -27.6 mV. MMT-Ser NC modified electrodes were evaluated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). Under the optimized conditions, working linear range for the modified sensor was found as 10-1000 µM with square wave voltammetry and the limit of detection (LOD) was found as 8 µM. The relative standard deviation for reproducibility was calculated as % 4.82 (n=6). Real sample analysis was performed with drug samples and the recovery values ranged from 94.64% to 111.2%.