Single-laboratory Study Research Articles

Determination of Total Aflatoxins in Polished Rice by Liquid Chromatography-Fluorescence Detection with Multifunctional Column Cleanup and Precolumn Derivatization: Single-Laboratory and Inter-Laboratory Validation Studies.

Aflatoxins (AFs) are toxic metabolites produced by Aspergillus spp. Because AFs are potent carcinogens in humans and animals, many countries have set regulatory limits for AFs in foods to prevent dietary exposure. From a global food safety perspective, in 2023, the Codex Alimentarius Commission established the maximum level (ML) of total AFs in certain cereals and cereal-based products, including polished rice. Therefore, validated analytical methods for AFs detection are necessary. In this study, an HPLC-fluorescence method coupled with multifunctional column cleanup and trifluoroacetic acid derivatization was developed for the determination of AFlevels in polished rice. Our method was validated in a single-laboratory study using AF-spiked materials, followed by an inter-laboratory validation study. Twelve laboratories participated in the inter-laboratory validation study, and five polished rice test samples artificially contaminated with AFswere analyzed. In a single-laboratory study, the ranges of mean recoveries of AFB1, B2, G1, G2, and total AFswere 101, 100-103, 93-96, 95-98, and 97-99%, respectively. The RSDs for within-day and between-day variations were all ≤4.4%. In the inter-laboratory validation study, the RSDs for repeatability and reproducibility were from 0.7 to 2.7% and 3.3 to 8.9% for all analytes, respectively. In response to the Codex ML and method performance criteria for AFsin polished rice, an analytical method based on HPLC-fluorescence detection was developed. All method performance parameters estimated from the test results of the single-laboratory and inter-laboratory validation studies met the criteria required by the Codex. Single- and inter-laboratory studies for the validation of an analytical method for AFlevel determination in polished rice were successfully performed. This analytical method will be suitable to determine AFlevels around the Codex ML set for polished rice.

Rapid identification and quantitation of 34 synthetic colorants in 125 beverages and candies by ultraperformance liquid chromatography with photodiode array detection

Ultraperformance liquid chromatography with photodiode array detection was used to simultaneously quantify 34 permitted and prohibited synthetic colorants in beverages and candies. Solvent extraction was employed to extract colorants from food samples, proving to be simpler, more rapid and cost-effective, and less labor-intensive than other pretreatment techniques. Additionally, a single-laboratory study was conducted to validate the above method. The results of this validation, which was performed using blank beverage and candy samples, demonstrated satisfactory linearity and detection/quantitation limits. According to the FDA guidelines for the validation of chemical methods, the method exhibited a recovery of 85.4–108 % and repeatability of 0.427–6.73 % for intra- and interday tests. The established method enabled the simultaneous analysis of 34 permitted and prohibited colorants within a short timeframe of 13 min, and its applicability was confirmed by analyzing 93 types of beverages and 32 types of candies available on the market to ensure that the colorant content in all products was within permissible limits. Thus, the developed method was concluded to be particularly suitable for the rapid, accurate, and convenient monitoring of colorant content in commercial products.

Determination of Acrylamide in Coffee, Cereals, Baby Food, Cocoa, Dry Pet Food, Potato Products, Vegetable Crisps, Biscuits, Tea, Nuts, and Spices by LC-MS/MS in a Single-Laboratory Validation: First Action 2023.01.

Acrylamide (AA) is a process contaminant naturally formed during the cooking of starchy food at high temperatures. Considering existing risks of misquantification inherent to the analysis of AA, an AOAC initiative raised the need for a consensus standard to determine AA in a broad variety of food. A quantitative LC-MS/MS method for AA determination in food was validated in a single-laboratory study. Targeted performance requirements in terms of target matrixes, limit of quantification, recovery, and precision were as defined per Standard Method Performance Requirement (SMPR®) 2022.006. The proposed method derives from EN 16618:2015 standard pending modifications brought to the (1) sample preparation (simplified, potentially automated); (2) scope of application (significantly extended); and (3) LC conditions (improved selectivity). Confirmatory detection of AA is conducted by LC-MS/MS in the Selected Reaction Monitoring mode (SRM), and isotopic dilution was applied for quantification approach using either 2,3,3-d3-acrylamide (d3-AA), or 13C3-2,3,3-d3-acrylamide (13C3-d3-AA) as labeled internal standard. A total of 16 laboratory samples from nine matrix categories were included in the validation process. A full validation was conducted on coffee (instant, roast), infant cereal, cocoa powder, pet food (croquettes), tea (green tea), spices (black pepper), and nuts (roasted almonds) with satisfactory performances both in terms of recovery (97-108%) and precision (RSDr and RSDiR <12%). The method applicability was further demonstrated through the analysis of quality control materials and reference materials including French fries, potato crisps, vegetable crisps, instant coffee, infant food, and biscuits (cookies), with accuracy values determined within a 94-107% range. The performances of the presented method are in agreement with the acceptance criteria stipulated in SMPR 2022.006. The Expert Review Panel for acrylamide approved the present method as AOAC Official First Action 2023.01.

Exploring δ13 C data compatibility in a broad δ13 C range: Reassembling δ13 C values obtained for NIST CO2 RMs 8562-8564 at different laboratories.

Stable isotope datasets obtained in different laboratories over years must be compared and combined. The strict compatibility in δ13 C at ±0.01‰ and ±0.02‰, k= 1, respectively, is required for observations of greenhouse gases CO2 and methane. That implies the long-term compatibility of calibrations realized at different laboratories. Thus far, this level of compatibility has not been demonstrated for pure CO2 in a broad δ13 C range and over years. We reassemble the δ13 C obtained in different laboratories for NIST CO2 RMs 8562-8564, which are pure CO2 gases. If the measurements are based on independent calibrations and reported on the same 17 O correction framework, δ13 C of all three RMs can be used to estimate the long-term δ13 C data compatibility. δ13 C values obtained in different laboratories for NIST CO2 RMs 8562-8564 (a range of δ13 C from ~ -3.7‰ to ~ -41.6‰) from 1998 to 2020 agree within their uncertainties. Two NIST studies (1999 and 2004) have combined data from several laboratories using different Isotope ratio mass spectrometry (IRMS) methods and somewhat different approaches. Three single-laboratory studies (2003, 2008, and 2020) have used the same type of IRMS and similar approaches. The δ13 C values from these laboratories agree within ±0.01‰, k= 1, over the entire δ13 C range and have uncertainties about two times lower than those obtained at NIST by combining data from several laboratories. The δ13 C values for NIST CO2 RMs 8562-8564 obtained in three laboratories using the same IRMS (2003, 2008, and 2020) agree within ±0.01‰, k= 1. First, this confirms the long-term stability in δ13 C of RMs 8562-8564. Second, this example of compatibility in δ13 C calibrations on pure CO2 , from -3.7‰ to -41.6‰, is relevant to stable isotope measurements of greenhouse gases CO2 and methane. Third, this confirms the high accuracy of the δ13 C(air-CO2 ) calibrations performed in 2003 and 2008 for the project Civil Aircraft for the Regular Investigation of the atmosphere Based on an Instrumented Container (CARIBIC).

Using mice from different breeding sites fails to improve replicability of results from single-laboratory studies

Theoretical and empirical evidence indicates that low external validity due to rigorous standardization of study populations is a cause of poor replicability in animal research. Here we report a multi-laboratory study aimed at investigating whether heterogenization of study populations by using animals from different breeding sites increases the replicability of results from single-laboratory studies. We used male C57BL/6J mice from six different breeding sites to test a standardized against a heterogenized (HET) study design in six independent replicate test laboratories. For the standardized design, each laboratory ordered mice from a single breeding site (each laboratory from a different one), while for the HET design, each laboratory ordered proportionate numbers of mice from the five remaining breeding sites. To test our hypothesis, we assessed 14 outcome variables, including body weight, behavioral measures obtained from a single session on an elevated plus maze, and clinical blood parameters. Both breeding site and test laboratory affected variation in outcome variables, but the effect of test laboratory was more pronounced for most outcome variables. Moreover, heterogenization of study populations by breeding site (HET) did not reduce variation in outcome variables between test laboratories, which was most likely due to the fact that breeding site had only little effect on variation in outcome variables, thereby limiting the scope for HET to reduce between-lab variation. We conclude that heterogenization of study populations by breeding site has limited capacity for improving the replicability of results from single-laboratory animal studies.

CRYPTOGENIC ORGANIZING PNEUMONIA: A DIAGNOSTIC DILEMMA IN A WORLD DOMINATED BY COVID-19 PNEUMONIA

CRYPTOGENIC ORGANIZING PNEUMONIA: A DIAGNOSTIC DILEMMA IN A WORLD DOMINATED BY COVID-19 PNEUMONIA

Establishing a multicenter, preclinical consortium in resuscitation: A pilot experimental trial evaluating epinephrine in cardiac arrest

BackgroundLarge animal studies are an important step in the translation pathway, but single laboratory experiments do not replicate the variability in patient populations. Our objective was to demonstrate the feasibility of performing a multicenter, preclinical, randomized, double-blinded, placebo-controlled cardiac arrest trial. We evaluated the effect of epinephrine on coronary perfusion pressure (CPP) as previous single laboratory studies have reported mixed results. MethodsForty-five swine from 5 different laboratories (Ann Arbor, MI; Baltimore, MD; Los Angeles, CA; Pittsburgh, PA; Toronto, ON) using a standard treatment protocol. Ventricular fibrillation was induced and left untreated for 6 min before starting continuous cardiopulmonary resuscitation (CPR). After 2 min of CPR, 9 animals from each lab were randomized to 1 of 3 interventions given over 12 minutes: (1) Continuous IV epinephrine infusion (0.00375 mg/kg/min) with placebo IV normal saline (NS) boluses every 4 min, (2) Continuous placebo IV NS infusion with IV epinephrine boluses (0.015 mg/kg) every 4 min or (3) Placebo IV NS for both infusion and boluses. The primary outcome was mean CPP during the 12 mins of drug therapy. ResultsThere were no significant differences in mean CPP between the three groups: 14.4 ± 6.8 mmHg (epinephrine Infusion), 16.9 ± 5.9 mmHg (epinephrine bolus), and 14.4 ± 5.5 mmHg (placebo) (p = NS). Sensitivity analysis demonstrated inter-laboratory variability in the magnitude of the treatment effect (p = 0.004). ConclusionThis study demonstrated the feasibility of performing a multicenter, preclinical, randomized, double-blinded cardiac arrest trials. Standard dose epinephrine by bolus or continuous infusion did not increase coronary perfusion pressure during CPR when compared to placebo.

Improving reproducibility in animal research by splitting the study population into several \u2018mini-experiments\u2019

In light of the hotly discussed ‘reproducibility crisis’, a rethinking of current methodologies appears essential. Implementing multi-laboratory designs has been shown to enhance the external validity and hence the reproducibility of findings from animal research. We here aimed at proposing a new experimental strategy that transfers this logic into a single-laboratory setting. We systematically introduced heterogeneity into our study population by splitting an experiment into several ‘mini-experiments’ spread over different time points a few weeks apart. We hypothesised to observe improved reproducibility in such a ‘mini-experiment’ design in comparison to a conventionally standardised design, according to which all animals are tested at one specific point in time. By comparing both designs across independent replicates, we could indeed show that the use of such a ‘mini-experiment’ design improved the reproducibility and accurate detection of exemplary treatment effects (behavioural and physiological differences between four mouse strains) in about half of all investigated strain comparisons. Thus, we successfully implemented and empirically validated an easy-to-handle strategy to tackle poor reproducibility in single-laboratory studies. Since other experiments within different life science disciplines share the main characteristics with the investigation reported here, these studies are likely to also benefit from this approach.

Determination of Lactose in Lactose-Free and Low-Lactose Milk, Milk Products, and Products Containing Dairy Ingredients by the LactoSens®R Amperometry Method: First Action 2020.01.

The AOAC Stakeholder Panel on Strategic Food Analytical Methods approved Standard Method Performance Requirements (SMPR®) 2018.009 for lactose in low-lactose or lactose-free milk, milk products, and products containing dairy ingredients. The LactoSens®R Method is a biosensor assay kit developed for the determination of lactose in a variety of lactose-free or low-lactose milk, dairy, and infant formula products produced with yeast-neutral lactases. In response to a call for methods, the LactoSensR method was validated in a single laboratory study with comparison to SMPR 2018.009. The LactoSensR method was evaluated for calibration, interference, repeatability, recovery, and robustness. In a method comparison study samples naturally containing low levels of lactose were evaluated using LactoSensR and an accredited high-performance anion-exchange chromatography with pulsed amperometric detection. Calibration with lactose standard solutions was shown to be linear and the method was shown to be free of interference from a variety of sugars, vitamins, alcohols, flavorings, and other compounds. Matrix studies, including 85 spiked materials, 55 products naturally containing lactose, and 13 reference materials, resulted in RSDr of 0-10.5% at 8-100 mg lactose/100 g and 0.2-5.4% at >100 mg lactose/100 g for milk and dairy products and 1.0-6.8% for infant formula, in compliance with SMPR 2018.009 with few exceptions. Recovery was 85.0-110.3% at 8-100 mg lactose/100 g and 85.6-109.7% at >100 mg lactose/100 g for milk and dairy products and 91.1-97.0% for infant formula, also meeting the performance requirements with few exceptions. The method was shown to be robust to changes in ambient temperature, sample temperature, and sample volume. The LactoSensR method compares favorably with the requirements of SMPR 2018.009 and should be adopted as a First Action AOAC Official MethodSM. The LactoSensR method is a fast, easy-to-use method that meets the requirements of SMPR 2018.009.

The dog as a sentinel species for environmental effects on human fertility.

Despite the vast body of evidence that environmental toxicants adversely affect reproductive development and function across species, demonstrating true cause and effect in the human remains a challenge. Human meta-analytical data, showing a temporal decline in male sperm quality, are paralleled by a single laboratory study showing a similar 26-year decline in the dog, which shares the same environment. These data are indicative of a common cause. Environmental chemicals (ECs) detected in reproductive tissues and fluids induce similar, short term, adverse effects on human and dog sperm. Both pre- and post-natal stages of early life development are sensitive to chemical exposures and such changes could potentially cause long term effects in the adult. The environmental 'pollutome' (mixtures of ECs) is determined by industrialisation, atmospheric deposition and bioaccumulation and characterises real-life exposure. In Arctic ecosystems, dietary and non-dietary chemical contaminants are detectable in biological tissues and linked with adverse health effects in both dogs and their handlers. In the female, such exposure could contribute to disorders such as ovarian insufficiency, dysregulated follicle development, ovarian cancer, and polycystic ovarian syndrome. In the dog, ovarian chemical concentrations are greater in the testis. In addition, preliminary studies indicate that dietary exposures may influence the sex ratio in the offspring in favour of females. Within this article, we review current knowledge on chemical effects on human reproduction and suggest that the dog, as a sentinel species for such effects, is an essential tool for addressing critical data gaps in this field.

Heterogenising study samples across testing time improves reproducibility of behavioural data

The ongoing debate on the reproducibility crisis in the life sciences highlights the need for a rethinking of current methodologies. Since the trend towards ever more standardised experiments is at risk of causing highly idiosyncratic results, an alternative approach has been suggested to improve the robustness of findings, particularly from animal experiments. This concept, referred to as “systematic heterogenisation”, postulates increased external validity and hence, improved reproducibility by introducing variation systematically into a single experiment. However, the implementation of this concept in practice requires the identification of suitable heterogenisation factors. Here we show that the time of day at which experiments are conducted has a significant impact on the reproducibility of behavioural differences between two mouse strains, C57BL/6J and DBA/2N. Specifically, we found remarkably varying strain effects on anxiety, exploration, and learning, depending on the testing time, i.e. morning, noon or afternoon. In a follow-up simulation approach, we demonstrate that the systematic inclusion of two different testing times significantly improved reproducibility between replicate experiments. Our results emphasise the potential of time as an effective and easy-to-handle heterogenisation factor for single-laboratory studies. Its systematic variation likely improves reproducibility of research findings and hence contributes to a fundamental issue of experimental design and conduct in laboratory animal science.

Reproducibility of preclinical animal research improves with heterogeneity of study samples.

Single-laboratory studies conducted under highly standardized conditions are the gold standard in preclinical animal research. Using simulations based on 440 preclinical studies across 13 different interventions in animal models of stroke, myocardial infarction, and breast cancer, we compared the accuracy of effect size estimates between single-laboratory and multi-laboratory study designs. Single-laboratory studies generally failed to predict effect size accurately, and larger sample sizes rendered effect size estimates even less accurate. By contrast, multi-laboratory designs including as few as 2 to 4 laboratories increased coverage probability by up to 42 percentage points without a need for larger sample sizes. These findings demonstrate that within-study standardization is a major cause of poor reproducibility. More representative study samples are required to improve the external validity and reproducibility of preclinical animal research and to prevent wasting animals and resources for inconclusive research.

The Exeter Trauma Stem: A radiographic follow-up at minimum of five years post implantation

IntroductionThe Exeter Trauma Stem (ETS) has been recommended by National Institute of Clinical Excellence (NICE) guidelines in the United Kingdom as a proven, cemented stem. A single laboratory study in the literature has raised possible concerns about the polished finish of the ETS and subsequent potential for accelerated loosening although there is little clinical evidence to support or refute this. MethodsThe aim of this study was to assess clinical outcomes of the ETS at a minimum of five years post implantation. Primary outcomes were radiological loosening at a minimum of five years along with survivorship of the implant. Patient demographics were prospectively collected and followed up. Results218 ETS’s (in 214 patients) were implanted from June 2002 until August 2008 in a single centre by a wide variety of surgeons of differing grades. Of these, 16 underwent revision surgery for fracture (2), dislocation (3), infection (1) and acetabular erosion (10) but there were no revisions for aseptic loosening of the implant. There were 64.0% (137/214) patients that had died by the time of this study. Of the remaining patients, 90 had radiographs of their hips at a minimum of 5 years with 36 of these at a minimum of 7 years post implantation. None of these had evidence of loosening. ConclusionThe ETS is a robust and suitable stem for implantation in patients with hip fractures. There are no clinical suspicions or increased rates of loosening with the ETS in our study. The concerns about surface finish are not borne out in our clinical study which shows no evidence of loosening at a minimum of five years post operation. It confers many advantages including ease of revision and it should continue to be used as per NICE guidelines.

Guidelines for the Statistical Analysis of a Collaborative Study of a Laboratory Method for Testing Disinfectant Product Performance

This paper presents statistical techniques suitable for analyzing a collaborative study (multilaboratory study or ring trial) of a laboratory disinfectant product performance test (DPPT) method. Emphasis is on the assessment of the repeatability, reproducibility, resemblance, and responsiveness of the DPPT method. The suggested statistical techniques are easily modified for application to a single laboratory study. The presentation includes descriptions of the plots and tables that should be constructed during initial examination of the data, including a discussion of outliers and QA checks. The statistical recommendations deal with evaluations of prevailing types of DPPTs, including both quantitative and semiquantitative tests. The presentation emphasizes tests in which the disinfectant treatment is applied to surface-associated microbes and the outcome is a viable cell count; however, the statistical guidelines are appropriate for suspension tests and other test systems. The recommendations also are suitable for disinfectant tests using any microbe (vegetative bacteria, virus, spores, etc.) or any disinfectant treatment. The descriptions of the statistical techniques include either examples of calculations based on published data or citations to published calculations. Computer code is provided in an appendix.

Evaluation of variability and quality control procedures for a receptor-binding assay for paralytic shellfish poisoning toxins

The receptor-binding assay (RBA) method for determining saxatoxin (STX) and its numerous analogues, which cause paralytic shellfish poisoning (PSP) in humans, was evaluated in a single laboratory study. Each step of the assay preparation procedure including the performance of the multi-detector TopCount® instrument was evaluated for its contribution to method variability. The overall inherent RBA variability was determined to be 17%. Variability within the 12 detectors was observed; however, there was no reproducible pattern in detector performance. This observed variability among detectors could be attributed to other factors, such as pipetting errors. In an attempt to reduce the number of plates rejected due to excessive variability in the method's quality control parameters, a statistical approach was evaluated using either Grubbs’ test or the Student's t-test for rejecting outliers in the measurement of triplicate wells. This approach improved the ratio of accepted versus rejected plates, saving cost and time for rerunning the assay. However, the potential reduction in accuracy and the lack of improvement in precision suggests caution when using this approach. The current study has recommended an alternate quality control procedure for accepting or rejecting plates in place of the criteria currently used in the published assay, or the alternative of outlier testing. The recommended procedure involves the development of control charts to monitor the critical parameters identified in the published method (QC sample, EC50, slope of calibration curve), with the addition of a fourth critical parameter which is the top value (100% binding) of the calibration curve.

フルオレスカミン誘導体化HPLC法による魚および水産加工品中のヒスタミン分析の性能評価

A method for the quantification of histamine in fish and fish products using tandem solid-phase extraction and fluorescence derivatization with fluorescamine was previously developed. In this study, we improved this analytical method to develop an official test method for quantification of histamine in fish and fish products, and performed a single laboratory study to validate it. Recovery tests of histamine from fillet (Thunnus obesus), and two fish products (fish sauce and salted and dried whole big-eye sardine) that were spiked at the level of 25 and 50 µg/g for T. obesus, and 50 and 100 µg/g for the two fish products, were carried out. The recoveries of histamine from the three samples tested were 88.8-99.6% with good repeatability (1.3-2.1%) and reproducibility (2.1-4.7%). Therefore, this method is acceptable for the quantification of histamine in fish and fish products. Moreover, surveillance of histamine content in food on the market was conducted using this method, and high levels of histamine were detected in some fish products.

A protocol for the validation of qualitative methods of detection

This paper presents a draft protocol for analyzing the results of validation studies for qualitative methods of detection which is designed to meet three competing goals: (1) to give correct answers, (2) have a broad scope of application, and (3) be accessible to a wide range of users. The draft protocol can be applied to the validation of methods by collaborative trial or to single-laboratory studies. The protocol produces an estimate of the probability of a positive response with a prediction interval within which 95% of laboratories (or analytical runs) are expected to fall when the method is applied in practice. The interval is calculated from the observed reproducibility (or within-laboratory reproducibility) standard deviation. Then a simple plot of prediction intervals for the probability of detection against the concentration of analyte, where the concentration is known, is used to provide an estimate of the range of limits of detection and false positive probability that we can expect to see when the validated method is applied in practice. The use of the draft protocol is demonstrated using results produced by three collaborative trials. A simulation study showed that a conclusion that a method is fit for purpose that is generated by the draft protocol is likely to be safe.

A meta-analytical estimation of the detection limits of methods for Salmonella in food

The validation of microbial detection methods for foods does not typically state a limit of detection value. Therefore performances of rapid and reference methods for Salmonella detection in foods were compared retrospectively using data from 49 published studies. A total of 576 values for the limit of detection (LOD50) were calculated. The major scientific variables in these independent validation studies were food matrices, Salmonella serovars, and method types (reference, cultural and immunological and nucleic acid based). The basic design feature of the original studies was comparison of the performances of new methods with those of cultural reference methods. A major experimental design variable was the number of laboratories per study. There were 29 single laboratory studies with 20 replicates per level of Salmonella spiked into the food matrix. The 20 multi-laboratory (N≥10) studies had 5–6 replicates per level of Salmonella. The LOD50 values of the dataset had a mean value of 0.021MPNg−1 (standard deviation range (0.013–0.036) and the distribution of their logarithms was symmetrical about the logarithm of the mean if the outlier values were discounted. Eighty-nine percent of the values ranged from 0.01 to 0.04MPNg−1. On this basis about 11% of the values were deemed outliers. About three-quarters of them were >0.04MPNg−1. The distribution derived in this study can be used as a bench mark against which to evaluate LOD50 data generated in future studies of detection methods for Salmonella in foods and potentially offers a possible way to streamline method validation study experimental design.

Probability of Detection (POD) as a Statistical Model for the Validation of Qualitative Methods

A statistical model is presented for use in validation of qualitative methods. This model, termed Probability of Detection (POD), harmonizes the statistical concepts and parameters between quantitative and qualitative method validation. POD characterizes method response with respect to concentration as a continuous variable. The POD model provides a tool for graphical representation of response curves for qualitative methods. In addition, the model allows comparisons between candidate and reference methods, and provides calculations of repeatability, reproducibility, and laboratory effects from collaborative study data. Single laboratory study and collaborative study examples are given.

Method performance study for total solids and total fat in coconut milk and products

Many analytical methods are available for total solids and total fat in various kinds of food. However, none is specific for coconut milk and products. The aim of this study was to establish methods and to conduct the method performance study on determination of total solids and total fat in coconut milk and products. In the single laboratory study, three methods for total solids determination were conducted: drying in hot air oven at 102±2°C and at 130±3°C and drying in vacuum oven at 70°C, 70mmHg, as well as three methods for total fat determination: direct extraction method, acid hydrolysis method and alkaline hydrolysis method. Sixteen samples of aqueous coconut milk and coconut milk powder were selected as test materials. Total solids and total fat contents were analyzed in 10 individual samples of each test material (N=10). The statistical analysis (one-way ANOVA) showed no significant difference for total solids and no difference for total fat content by the used methods (p>0.05). Since the drying method by hot air oven at 102±2°C is the most commonly used for total solids in food laboratories, it was selected for further inter-laboratory study. For total fat determination, the alkaline hydrolysis method (Roese–Gottlieb) was selected for further inter-laboratory study since it is less time-consuming and involves fewer steps than the acid hydrolysis method. Seventeen laboratories participated in the inter-laboratory study and seven representative test materials with varying levels of total solids and fat were used. The method performance characteristics are presented. These methods were proposed to CODEX and endorsed as the CODEX standard methods (type I) for aqueous coconut milk and coconut cream analyses.