Standardization of chromatographic signals - Part I: Towards obtaining instrument-agnostic fingerprints in gas chromatography.

One of the main causes for the sparse use of multivariate analytical methods in routine laboratory work is the dependency on the measuring instrument from which the analytical signal is acquired. This issue is especially critical in chromatographic equipment and results in limitations of their applicability. The solution to this problem is to obtain a standardized instrument-independent signal -or instrument-agnostic signal- regardless of the measuring instrument or of the state of the same instrument from which it has been acquired. The combined use of both internal and external standard series, allows us to have external and transferable references for the normalization of both the intensity and the position of each element of the data vector being arranged from the raw signal. From this information, a simple mathematical data treatment process is applied and instrument-agnostic signals can be secured. This paper describes and applies the proposed methodology to be followed for obtaining standardized instrumental fingerprints from two significant fractions of virgin olive oil (volatile organic compounds and triacylglycerols), obtained by gas chromatography coupled to mass spectrometry (GC-MS) and analysed with two temperature conditions (conventional and high-temperature, respectively). The results of both case studies show how the instrument-agnostic fingerprints obtained are coincidental, regardless of the state of the chromatographic system or the time of acquisition.

ROC curves for the optimization of one-class model parameters. A case study: Authenticating extra virgin olive oil from a Catalan protected designation of origin.

This paper proposes a ROC curve-based methodology to find optimal classification model parameters. ROC curves are implemented to set the optimal number of PCs to build a one-class SIMCA model and to set the threshold class value that optimizes both the sensitivity and specificity of the model. The authentication of the geographical origin of extra-virgin olive oils of Arbequina botanical variety is presented. The model was developed for samples from Les Garrigues, target class, Samples from Siurana were used as the non-target class. Samples were measured by FT-Raman with no pretreatment. PCA was used as exploratory technique. Spectra underwent pre-treatment and variables were selected based on their VIP score values. ROC curve and others already known criteria were applied to set the threshold class value. The results were better when the ROC curve was used, obtaining performance values higher than 82%, 75% and 77% for sensitivity, specificity and efficiency, respectively.

The occurrence: A meaningful parameter to be considered in the validation of multivariate classification-based screening methods - Application for authenticating virgin olive oil.

The development of multivariate screening analytical methods in the analytical chemistry field focused particularly on food authentication is growing in recent years, which is evidenced by the increase of scientific publications. Currently there are several guides and technical reports about how -univariate qualitative methods should be properly validated to produce reliable and accurate (fitted-for-purpose) results. Nevertheless, this is not the case when multivariate methods are considered. Aimed at redressing this untenable disadvantage, this paper proposes some guidelines for the validation of multivariate classification-based screening methods. As an application example, the detection of adulteration of virgin olive oil with any other edible vegetal oils is showed. The analytical techniques employed are liquid chromatography coupled to diode array detector (LC-DAD) and gas chromatography coupled to flame ionization detector (GC-FID). For the correct validation of the multivariate screening method a new parameter which never considered before, named occurrence, is accounted. Also, it has been developed two new applicability indicators of the multivariate screening methods: the assignation error index (IERROR) and the index saving (ISAVING) to establish the validation requirements. Then the validation parameters of the methods: precision (or target predictive value), sensitivity, non-target predictive value, specificity and accuracy were estimated. The main conclusion of the work has been the need to take accounts the occurrence value to establish the specific validation requirements to apply the multivariate screening method in a particular scenario.

Authentication of the geographical origin of extra-virgin olive oil of the Arbequina cultivar by chromatographic fingerprinting and chemometrics.

This paper proposes to use chromatographic fingerprints coupled to multivariate techniques to authenticate the geographical origin of extra-virgin olive oils (EVOO) of the Arbequina botanical variety. This methodology uses the whole or part of the chromatogram as input data for the classification models but does not identify or quantify the chemicals constituents. Arbequina monovarietal EVOOs from three geographical origins were studied: two from adjacent European Protected Designation of Origin areas, Siurana and Les Garrigues, in Catalonia in the northeast of Spain; and the third from the south of Spain (Andalucia and Murcia). Three chromatographic fingerprints of each sample were obtained by both reverse and normal phase liquid chromatography coupled to charged aerosol detector (HPLC-CAD), and high temperature gas chromatography coupled to flame ionization detector [(HT)GC-FID]. Principal component analysis (PCA) was used as exploratory technique and soft independent modelling of class analogy (SIMCA) and partial least square-discriminant analysis (PLS-DA) were used as classification methods. High and low-level data fusion strategies were also applied to improve the classification results obtained when the data acquired from each analytical technique were separately used. The results were best for the PLS-DA model with low-level fusion of two techniques (HT)GC-FID with HPLC-CAD, independently of the phase mode. Sensitivity and specificity were 100% in almost all classes, error was 0% for all classes and an inconclusive ratio of just 4% was obtained for the Les Garrigues class due to double assignations.

HPLC-UV and HPLC-CAD chromatographic data fusion for the authentication of the geographical origin of palm oil.

Data fusion combined with a multivariate classification approach (partial least squares-discriminant analysis, PLS-DA) was applied to authenticate the geographical origin of palm oil. Data fusion takes advantage of the synergistic effect of information collected from more than one data source. In this study, data from liquid chromatography coupled to two detectors -ultraviolet (UV) and charged aerosol (CAD)- was fused by high- and mid-level data fusion strategies. Mid-level data fusion combines a few variables from each technique and then applies the classification technique. Principal component analysis and interval partial least squares were applied to obtain the variables selected. High-level data fusion combines the PLS-DA classification results obtained individually from the chromatographic technique with each detector. Fuzzy aggregation connective operators were used to make the combinations. Prediction rates varied between 73% and 98% for the individual techniques and between 87% and 100% and 93% and 100% for the mid- and high-level data fusion strategies, respectively.

A new analytical method for quantification of olive and palm oil in blends with other vegetable edible oils based on the chromatographic fingerprints from the methyl-transesterified fraction.

A new analytical method for the quantification of olive oil and palm oil in blends with other vegetable edible oils (canola, safflower, corn, peanut, seeds, grapeseed, linseed, sesame and soybean) using normal phase liquid chromatography, and applying chemometric tools was developed. The procedure for obtaining of chromatographic fingerprint from the methyl-transesterified fraction from each blend is described. The multivariate quantification methods used were Partial Least Square-Regression (PLS-R) and Support Vector Regression (SVR). The quantification results were evaluated by several parameters as the Root Mean Square Error of Validation (RMSEV), Mean Absolute Error of Validation (MAEV) and Median Absolute Error of Validation (MdAEV). It has to be highlighted that the new proposed analytical method, the chromatographic analysis takes only eight minutes and the results obtained showed the potential of this method and allowed quantification of mixtures of olive oil and palm oil with other vegetable oils.

Fast-HPLC Fingerprinting to Discriminate Olive Oil from Other Edible Vegetable Oils by Multivariate Classification Methods.

A new analytical method for the differentiation of olive oil from other vegetable oils using reversed-phase LC and applying chemometric techniques was developed. A 3 cm short column was used to obtain the chromatographic fingerprint of the methyl-transesterified fraction of each vegetable oil. The chromatographic analysis took only 4 min. The multivariate classification methods used were k-nearest neighbors, partial least-squares (PLS) discriminant analysis, one-class PLS, support vector machine classification, and soft independent modeling of class analogies. The discrimination of olive oil from other vegetable edible oils was evaluated by several classification quality metrics. Several strategies for the classification of the olive oil were used: one input-class, two input-class, and pseudo two input-class.

One input-class and two input-class classifications for differentiating olive oil from other edible vegetable oils by use of the normal-phase liquid chromatography fingerprint of the methyl-transesterified fraction.

A new method for differentiation of olive oil (independently of the quality category) from other vegetable oils (canola, safflower, corn, peanut, seeds, grapeseed, palm, linseed, sesame and soybean) has been developed. The analytical procedure for chromatographic fingerprinting of the methyl-transesterified fraction of each vegetable oil, using normal-phase liquid chromatography, is described and the chemometric strategies applied and discussed. Some chemometric methods, such as k-nearest neighbours (kNN), partial least squared-discriminant analysis (PLS-DA), support vector machine classification analysis (SVM-C), and soft independent modelling of class analogies (SIMCA), were applied to build classification models. Performance of the classification was evaluated and ranked using several classification quality metrics. The discriminant analysis, based on the use of one input-class, (plus a dummy class) was applied for the first time in this study.