Optimization of headspace high-capacity tool coupled to two-dimensional gas chromatography-mass spectrometry for mapping the volatile organic compounds of raw pistachios. A proof-of-concept on the classification ability by geographic origin.

An optimized procedure for extracting and analyzing raw pistachio volatiles was developed through headspace sampling with high-capacity tools and subsequent analysis using comprehensive two-dimensional gas chromatography coupled with mass spectrometry. The examination of 18 pistachio samples belonging to different geographic areas led to the identification of a set of 99 volatile organic compounds (VOCs). Molecules were putatively identified using linear retention index, mass spectra similarity, and two-dimensional plot location. The impact of preprocessing and processing techniques on the aligned data matrix from a set of samples of different geographical origins, after removing contaminants, was evaluated. The combination of scaling with log-transformation, normalization with z-score, and data reduction with random forest machine learning algorithm generated a panel of 16 discriminatory VOC molecules. As a proof of concept, raw pistachios' VOC profile was employed for the first time to tentatively classify them based on their geographical origin.

Microwave-Assisted Extraction Coupled to HPLC-UV Combined with Chemometrics for the Determination of Bioactive Compounds in Pistachio Nuts and the Guarantee of Quality and Authenticity.

Two novel microwave-assisted extraction (MAE) methods were developed for the isolation of phenols and tocopherols from pistachio nuts. The extracts were analyzed by reversed-phase high-pressure liquid chromatography coupled with a UV detector (RP-HPLC-UV). In total, eighteen pistachio samples, originating from Greece and Turkey, were analyzed and thirteen phenolic compounds, as well as α-tocopherol, (ß + γ)-tocopherol, and δ-tocopherol, were identified. The analytical methods were validated and presented good linearity (r2 > 0.990) and a high recovery rate over the range of 82.4 to 95.3% for phenols, and 93.1 to 96.4% for tocopherols. Repeatablility was calculated over the range 1.8-5.8%RSD for intra-day experiments, and reproducibility over the range 3.2-9.4%RSD for inter-day experiments, respectively. Principal component analysis (PCA) was employed to analyze the differences between the concentrations of the bioactive compounds with respect to geographical origin, while agglomerative hierarchical clustering (AHC) was used to cluster the samples based on their similarity and according to the geographical origin.

Genome-wide identification of Pistacia R2R3-MYB gene family and function characterization of PcMYB113 during autumn leaf coloration in Pistacia chinensis.

Pistacia chinensis is known for its biodiesel production. Several varieties of this plant have leaves that produce anthocyanin, which is responsible for their reddish coloration in autumn. This reddish hue is what makes them useful as ornamental plants. However, the mechanism of anthocyanin accumulation during autumn leaf coloration remains unclear. R2R3-MYB proteins reportedly regulated anthocyanin biosynthesis in many plant species. Here, we performed a genome-wide analysis and expression profiles of R2R3-MYB transcription factor in Pistacia. A total of 158 R2R3-MYB proteins were identified and grouped into 32 clades. Combining the data from RNA-seq and qRT-PCR, one key gene, EVM0016534, was screened and identified to have the highest correlation with anthocyanin accumulation. It was named PcMYB113 due to its sequence similarity to AtMYB113 and it could bind to the promoter of PcF3H. Furthermore, ectopic expression of PcMYB113 in Arabidopsis promoted the accumulation of anthocyanin in the seed coat, cotyledon, and mature leaves, thus confirming the function of PcMYB113 in anthocyanin biosynthesis. In addition, PcMYB113 had a specifically higher expression in senesced red leaves than in mature green leaves and young red leaves in P. chinensis, thereby suggesting the potential role of PcMYB113 in promoting anthocyanin biosynthesis during autumn leaf coloration. These findings enrich our understanding of the function of R2R3-MYB genes in anthocyanin biosynthesis and autumn leaf coloration.

Influence of cultivar and technological conditions on the volatile profile of virgin pistachio oils.

The main aim of this work was to characterize the volatile profile of virgin pistachio oils produced from eight cultivars (Aegina, Avdat, Kastel, Kerman, Larnaka, Mateur, Napoletana and Sirora), under different technological conditions (temperature, roasting, use of whole nuts, screw speed and nozzle diameter), and compare it with those of commercial pistachio oils. Terpenes (15.57-41.05 mg/kg), accounting for ~97% of total volatiles, were associated with appreciated sensory properties, with α-pinene as the main volatile (14.47-37.09 mg/kg). Other terpene compounds such as limonene (0.11-3.58 mg/kg), terpinolene (0.00-1.61 mg/kg), ß-pinene (0.12-1.20 mg/kg) and α-terpineol (0.00-1.17 mg/kg) were quantified at lower concentrations. Acids, alcohols, aldehydes, esters and hydrocarbons only summed to ~3% of the total volatile compounds. The volatiles content greatly depended on the pistachio cultivar employed. The influence of extraction conditions was also very relevant; in particular, terpenes doubled (28.38-53.84 mg/kg) using whole pistachios for oil extraction, also being incremented by mild processing conditions. On the contrary, higher temperature or roasting decreased the terpene content (~50-25% respectively), and pyrazines appeared (up to 3.12 mg/kg).

Functional and sensory properties of pistachio nuts as affected by cultivar.

BACKGROUND: Modern agriculture allows farmers to choose among different cultivars of the same fruit to fulfill their agronomic needs and consumers' demands; however, there are only a few studies that describe and compare key functional and sensory properties of different pistachio cultivars. The main objective of this study was to compare eight pistachio cultivars by analyzing key functional properties (phenolic compounds, polymeric procyanidins, antioxidant activity, and inhibition of α-amylase and ß-glucosidase), aromatic compounds with gas chromatography-mass spectrometry (GC-MS), and by examining their sensory properties with a trained panel. RESULTS: A combination of LC-PDA-MS-QTof (liquid chromatography photodiode array quadrupole time-off flight mass spectrometry) and electrospray ionization was used to determine two phenolic acids, nine flavonols, one anthocyanin, and three flavan-3-ols in pistachio cultivars, with a total concentration ranging from 500 to 6065 mg 100 g-1 dry weight (dw). The total polymeric procyanidins concentrations oscillated between 348 and 5919 mg 100 g-1 dw, (-)-epicatechin being the major monomer contributor. Pinene was the most abundant volatile compound (∼200 mg kg-1 dw), and, in the sensory analysis of samples, 23 sensory attributes were found to differ significantly among the cultivars. CONCLUSION: The cultivar 'Larnaka' stood out as having the best functional profile (high polyphenolic content, high antioxidant activity, and high values of α-amylase and ß-glucosidase inhibition), and the cultivars 'Kastel' and 'Kerman' showed the most attractive sensory properties, mainly the most intense flavor. © 2019 Society of Chemical Industry.

Identification and Pathogenicity of Fungal Species Associated with Canker Diseases of Pistachio in California.

A survey was conducted during 2015 and 2016 in pistachio orchards throughout the San Joaquin Valley of California to investigate the occurrence of canker diseases and identify the pathogens involved. Cankers and dieback symptoms were observed mainly in orchards aged >15 years. Symptoms of canker diseases included brown to dark brown discoloration of vascular tissues, wood necrosis, and branch dieback. In total, 58 fungal isolates were obtained from cankers and identified based on multilocus phylogenetic analyses (internal transcribed spacer, glyceraldehyde 3-phosphate dehydrogenase, ß-tubulin, calmodulin, actin 1, and translation elongation factor 1α) representing 11 fungal species: Colletotrichum karstii, Cytospora californica, Cytospora joaquinensis, Cytospora parapistaciae, Cytospora pistaciae, Diaporthe ambigua, Didymella glomerata, Diplodia mutila, Neofusicoccum mediterraneum, Phaeoacremonium canadense, and Schizophyllum commune. Pathogenicity tests conducted in the main pistachio cultivars Kerman, Golden Hills, and Lost Hills using the mycelium-plug method indicated that all fungal species were pathogenic to Pistacia vera. All species tested caused cankers in pistachio branches, although virulence among species varied from high to moderate. Overall, N. mediterraneum and Cytospora spp. were the most widespread and virulent species associated with canker diseases of pistachio in California.

Cumulative risk assessment of pesticide residues in different Iranian pistachio cultivars: Applying the source specific HQS and adversity specific HIA approaches in Real Life Risk Simulations (RLRS).

Iran is a major supplier of the world pistachio market. In this study, we collected five pistachio cultivars from four main pistachio-producing zones in August and September 2016, and determined the residues of 18 organophosphorus, carbamate, pyrethroid and nicotinoid pesticide in these samples using Gas Chromatography-Mass Spectrometry, as an efficient method for determination of pesticides' residues. Next, single-chemical and chronic cumulative risk assessment was done based on the new approaches of the food specific Hazard Quotient and adversity specific Hazard Index. Fifteen from eighteen food-specific Hazard Quotients were above 1 even in cases when the respective contamination was bellow MRLs. The adversity specific Hazard Indexes values were above 1 for five from six adversities indicating various risks in the resulted levels of pistachios' contamination from the pesticides' mixture. However, no risk for carcinogenicity was found. Our results indicate the necessity of taking appropriate measures to control/standardize pesticides practice in pistachio cultivation in Iran and the need to re-establish the MRLs based on cumulative exposure.

Comprehensive Study of the Phenolic Compound Profile and Antioxidant Activity of Eight Pistachio Cultivars and Their Residual Cakes and Virgin Oils.

Phenolic compounds of eight pistachio ( Pistacia vera L.) cultivars and their residual cakes and virgin oils (screw pressing) were studied using high-performance liquid chromatography-diode array detection-electrospray ionization-tandem mass spectrometry. A total of 25 compounds were identified and quantified for pistachio nuts and residual cakes, with the presence of five flavonols, six flavanols, and one gallotannin being reported for the first time. Total phenolics in pistachio nuts showed a concentration from 1359 mg/kg (Kastel) to 4507 mg/kg (Larnaka). Flavanols were the most abundant phenolics, at about 90%, with resulting procyanidin B1 and gallocatechin being the main phenolics, depending upon the cultivar. Other phenolic groups, such as anthocyanins (from 54 to 218 mg/kg), flavonols (from 76 to 130 mg/kg), flavanones (from 12 to 71 mg/kg), and gallotannins (from 4 to 46 mg/kg), were also identified. Residual cakes presented the same phenolic profile but with a concentration almost double because of the concentration effect caused by the oil separation. Virgin pistachio oils showed a very low phenolic content, with eriodyctiol being the only compound identified.

Chemical partitioning and DNA fingerprinting of some pistachio (Pistacia vera L.) varieties of different geographical origin.

The genus Pistacia (Anacardiaceae family) is represented by several species, of which only P. vera L. produces edible seeds (pistachio). Despite the different flavor and taste, a correct identification of pistachio varieties based on the sole phenotypic character is sometimes hard to achieve. Here we used a combination of chemical partitioning and molecular fingerprinting for the unequivocal identification of commercial pistachio seed varieties (Bronte, Kern, Kerman, Larnaka, Mateur and Mawardi) of different geographical origin. The total phenolic content was higher in the variety Bronte followed by Larnaka and Mawardi cultivars. The total anthocyanin content was higher in Bronte and Larnaka varieties, whereas the total proanthocyanidin content was higher in Bronte, followed by Mawardi and Larnaka varieties. HPLC-DAD-ESI-MS/MS analyses revealed significant (P < 0.05) higher amounts of cyanidin-3-glucoside, idein, eryodictol-7-galactoside, quercetin-3-glucoside, luteolin-glucoside and marein in the variety Bronte, whereas higher amounts of peonidin-3-glucoside, okanin 4'-galactoside, hyperoside and quercetin-4'-glucoside were found in the variety Larnaka. The highest content of catechin was found in the Mawardi variety. A significantly (P < 0.05) higher total amount of fatty acids was found in the varieties Mateur, Kern and Bronte, followed by the varieties Larnaka and Mawardi, whereas the variety Kerman showed the lowest total fatty acid content. GC-FID and GC-MS analyses revealed the presence of several polyunsaturated fatty acids. Kern and Mateur varieties showed a significantly (P < 0.05) higher amount of linoleic acid, whereas the variety Bronte showed the highest amount of oleic acid. Molecular fingerprinting was achieved by ITS DNA PCR-RFLP analysis. Three different restriction enzymes (RsaI, TaqαI and PstI) were used to selectively cleave the resulting amplicons. A TaqαI site could be selectively found in the varieties Kerman, Larnaka and Mateur, whereas the digestion of the PCR products by RsaI gave specific patters exclusively on Bronte and Mawardi. Digestion by PstI gave specific patters exclusively on the Kern variety. The results showed that the Mediterranean varieties (Mateur, Bronte and Larnaka) show similar chemical patterns and (particularly for Mateur and Larnaka) a close phylogenetic relationship, allowing a chemical and molecular partitioning with respect to the other varieties.

Discovery of Viruses and Virus-Like Pathogens in Pistachio using High-Throughput Sequencing.

Pistachio (Pistacia vera L.) trees from the National Clonal Germplasm Repository (NCGR) and orchards in California were surveyed for viruses and virus-like agents by high-throughput sequencing (HTS). Analyses of sequence information from 60 trees identified a novel virus, provisionally named "Pistachio ampelovirus A" (PAVA), in the NCGR that showed low amino acid sequence identity (approximately 42%) compared with members of the genus Ampelovirus (family Closteroviridae). A putative viroid, provisionally named "Citrus bark cracking viroid-pistachio" (CBCVd-pis), was also found in the NCGR and showed approximately 87% similarity to Citrus bark cracking viroid (CBCVd, genus Cocadviroid, family Pospiviroidae). Both PAVA and CBCVd-pis were graft transmissible to healthy UCB-1 hybrid rootstock seedlings (P. atlantica × P. integerrima). A field survey of 123 trees from commercial orchards found no incidence of PAVA but five (4%) samples were infected with CBCVd-pis. Of 675 NCGR trees, 16 (2.3%) were positive for PAVA and 172 (25.4%) were positive for CBCVd-pis by reverse-transcription polymerase chain reaction. Additionally, several contigs across multiple samples exhibited significant sequence similarity to a number of other plant virus species in different families. These findings require further study and confirmation. This study establishes the occurrence of viral and viroid populations infecting pistachio trees.

Chemical and biomolecular analyses to discriminate three taxa of Pistacia genus from Sardinia Island (Italy) and their antifungal activity.

This work reports the results and the comparison concerning the chemical and biomolecular analyses and the antifungal activity of three wild Pistacia species (Anacardiaceae) from Sardinia. Volatile oils from leaves and twigs of Pistacia x saportae, Pistacia lentiscus and Pistacia terebinthus were characterised using GC-FID and GC-MS techniques and tested against some fungal strains. Two DNA nuclear regions (ITS and 5S-rRNA-NTS) were amplified through PCR technique and sequenced. The three **Pistacia have similar chemical profile, although there are some important quantitative differences. The analysis of ITS and 5S-rRNA-NTS regions, reveals a species-specific nucleotide variation among the three **taxa. This method could emerge as a powerful tool for the species identification, especially because the discrimination of these three **taxa appears difficult for non-expert botanists. Concerning the antifungal activity, P. lentiscus and P. x saportae show the highest activity against Cryptococcus neoformans, with a MIC value of 0.32 µL/mL.

Health risk assessment of heavy metals via dietary intake of five pistachio (Pistacia vera L.) cultivars collected from different geographical sites of Iran.

Pistachio is an important horticultural product and Iran is considered as a main pistachio producing country. Assessment of heavy metals in this export fruit is crucial for protecting public health against toxic heavy metals. The concentration of selected heavy metals in soil, water and five pistachio cultivars from four geographical regions of Iran were measured. Although none of the elements were detected in water irrigation, infield metal content in the soil had good correlation with that of pistachio. The highest amounts of Al, As, Co, Ni and Se were reported in samples collected from Sarakhs, Iran. Considering both cultivar and region effects on selected heavy metals concentration, Kaleghoochi cultivar from Sarakhs site showed the highest amount of Al, As, Ni and Se. The maximum concentration of Hg was found in Akbari cultivar collected from Damghan. In the Akbari and the Ahmad aghaei cultivars collected from Sarakhs and Damghan cultivation zones, respectively, the highest amount of Co were observed. Based on our results, the HI value for the consumers of Iranian pistachio was 0.066. It seems that the levels of heavy metals in these pistachio samples pose no risk to consumers.

Genome survey of pistachio (Pistacia vera L.) by next generation sequencing: Development of novel SSR markers and genetic diversity in Pistacia species.

BACKGROUND: Pistachio (Pistacia vera L.) is one of the most important nut crops in the world. There are about 11 wild species in the genus Pistacia, and they have importance as rootstock seed sources for cultivated P. vera and forest trees. Published information on the pistachio genome is limited. Therefore, a genome survey is necessary to obtain knowledge on the genome structure of pistachio by next generation sequencing. Simple sequence repeat (SSR) markers are useful tools for germplasm characterization, genetic diversity analysis, and genetic linkage mapping, and may help to elucidate genetic relationships among pistachio cultivars and species. RESULTS: To explore the genome structure of pistachio, a genome survey was performed using the Illumina platform at approximately 40× coverage depth in the P. vera cv. Siirt. The K-mer analysis indicated that pistachio has a genome that is about 600 Mb in size and is highly heterozygous. The assembly of 26.77 Gb Illumina data produced 27,069 scaffolds at N50 = 3.4 kb with a total of 513.5 Mb. A total of 59,280 SSR motifs were detected with a frequency of 8.67 kb. A total of 206 SSRs were used to characterize 24 P. vera cultivars and 20 wild Pistacia genotypes (four genotypes from each five wild Pistacia species) belonging to P. atlantica, P. integerrima, P. chinenesis, P. terebinthus, and P. lentiscus genotypes. Overall 135 SSR loci amplified in all 44 cultivars and genotypes, 41 were polymorphic in six Pistacia species. The novel SSR loci developed from cultivated pistachio were highly transferable to wild Pistacia species. CONCLUSIONS: The results from a genome survey of pistachio suggest that the genome size of pistachio is about 600 Mb with a high heterozygosity rate. This information will help to design whole genome sequencing strategies for pistachio. The newly developed novel polymorphic SSRs in this study may help germplasm characterization, genetic diversity, and genetic linkage mapping studies in the genus Pistacia.

Identification of Phenolic Compounds in Red and Green Pistachio (Pistacia vera L.) Hulls (Exo- and Mesocarp) by HPLC-DAD-ESI-(HR)-MS(n).

Phenolic constituents of the nonlignified red and green pistachio hulls (exo- and mesocarp) were assessed by HPLC-DAD-ESI-MS(n) as well as by HR-MS. A total of 66 compounds was identified in the respective aqueous methanolic extracts. Among them, gallic acid, monogalloyl glucoside, monogalloyl quinic acid, penta-O-galloyl-ß-d-glucose, hexagalloyl hexose, quercetin 3-O-galactoside, quercetin 3-O-glucoside, quercetin 3-O-glucuronide, and (17:1)-, (13:0)-, and (13:1)-anacardic acids were detected at highest signal intensity. The main difference between red and green hulls was the presence of anthocyanins in the former ones. Differently galloylated hydrolyzable tannins, anthocyanins, and minor anacardic acids were identified for the first time. Pistachio hulls were thus shown to be a source of structurally diverse and potentially bioactive phenolic compounds. They therefore represent a valuable byproduct of pistachio processing having potential for further utilization as raw material for the recovery of pharmaceutical, nutraceutical, and chemical products.

Potential Start Codon Targeted (SCoT) and Inter-retrotransposon Amplified Polymorphism (IRAP) Markers for Evaluation of Genetic Diversity and Conservation of Wild Pistacia Species Population.

Wild pistachio species is important species in forests regions Iran and provide protection wind and soil erosion. Even though cultivation and utilization of Pistacia are fully exploited, the evolutionary history of the Pistacia genus and the relationships among the species and accessions is still not well understood. Two molecular marker strategies, SCoT and IRAP markers were analyzed for assessment of 50 accessions of this species accumulated from diverse geographical areas of Iran. A thorough of 115 bands were amplified using eight IRAP primers, of which 104 (90.4 %) have been polymorphic, and 246 polymorphic bands (68.7 %) had been located in 358 bands amplified by way of forty-four SCoT primers. Average PIC for IRAP and SCoT markers became 0.32 and 0.48, respectively. This is exposed that SCoT markers have been extra informative than IRAP for the assessment of variety among pistachio accessions. Primarily based on the two extraordinary molecular markers, cluster evaluation revealed that the 50 accessions taken for the evaluation may be divided into three distinct clusters. Those results recommend that the performance of SCoT and IRAP markers was highly the equal in fingerprinting of accessions. The results affirmed a low genetic differentiation among populations, indicating the opportunity of gene drift most of the studied populations. These findings might render striking information in breeding management strategies for genetic conservation and cultivar improvement.

Tempo de destilação e perfil volátil do óleo essencial de aroeira da praia (Schinus terebinthifolius) em Sergipe / Distillation time and volatile profile of the essential oil of Brazilian pepper (Schinus terebinthifolius) in Sergipe, Brazil

Aos frutos e óleos essenciais da aroeira da praia (Schinus terebinthifolius) atribui-se atividade antimicrobiana sobre bactérias gram positivas além de ação anti-inflamatória. O óleo essencial da aroeira é utilizado no tratamento de micoses, sendo a sua atividade antifúngica atribuída à alta concentração de monoterpenos. Este estudo teve como objetivo determinar o melhor rendimento de destilação de óleo essencial de sementes e folhas da aroeira e os componentes químicos do óleo essencial nos diversos períodos de destilação. Cem gramas de cada material foram destilados em aparato de Clevenger durante 2,5h, 4,0h, 5,5h e 7,0h. O material destilado foi analisado por cromatografia a gás acoplada a espectrofotômetro de massas, onde foi obtido o perfil cromatográfico do material. As sementes de aroeira apresentaram maiores quantidades de óleo essencial, quando comparadas com as folhas em todos os períodos de destilação. O óleo essencial de aroeira mostrou perfil volátil formado principalmente por mono e sesquiterpenos.

The fruits and essential oils of Brazilian pepper (Schinus terebinthifolius) have antimicrobial activity against gram-positive bacteria, as well as anti-inflammatory action. The essential oil of Brazilian pepper is used to treat fungal infections, and its antifungal activity is attributed to the high concentration of monoterpenes. This study aimed to determine the best yield of distillation of the essential oil of seeds and leaves of Brazilian pepper and the chemical components of the essential oil in different periods of distillation. One hundred grams of each material were distilled in Clevenger apparatus for 2.5 h, 4.0 h, 5.5 h and 7.0 h. The distilled material was analyzed by gas chromatography coupled to mass spectrometer, where the chromatographic profile of the material was obtained. Seeds of Brazilian pepper showed higher amounts of essential oils when compared with the leaves in all the periods of distillation. The essential oil of Brazilian pepper showed a profile consisting mainly of volatile mono- and sesquiterpenes.

Survey of ex situ fruit and leaf volatiles from several Pistacia cultivars grown in California.

BACKGROUND: California is the second largest cultivator of pistachios, producing over 375 million pounds and a revenue of $787 million in 2009. Despite the agricultural and economic importance of pistachios, little is known regarding their actual volatile emissions, which are of interest owing to their potential roles as semiochemicals to insect pests. RESULTS: The ex situ volatile analysis of leaves from Pistacia atlantica, P. chinensis, P. lentiscus, P. palaestina, P. terebinthus, P. vera and P. weimannifolia demonstrated emission differences between species as well as between female and male leaves. Leaves from the female P. vera cultivars Bronte, Damghan, II, III, Kerman and Ohadi as well as fruits of P. atlantica, P. chinensis, P. lentiscus, P. palaestina, P. terebinthus and P. vera (cultivars II, III, Kaleh, Kerman, Momtaz and Ohadi) showed differences in the composition and relative quantity of major volatiles. The compounds in highest relative quantities from the various analyses were sabinene, Δ(3)-carene, ß-myrcene, α-phellandrene, limonene, (Z)-ocimene, (E)-ß-ocimene and α-terpinolene. CONCLUSION: This is the first ex situ survey of fruit and leaf volatile emissions from California-grown Pistacia species and a number of corresponding cultivars. The study provides an overview of the major and minor volatile emissions and also offers evidence of chemotypes based on monoterpenes. The results highlight the dissimilarity of major components detected between ex situ volatile collection and essential oil analysis.

Use of chemical profiling to differentiate geographic growing origin of raw pistachios.

The objective of this study was to demonstrate the feasibility of chemical profiling methods combined with multivariate methods to differentiate the geographical growing regions of pistachios (Pistachia vera). Elemental analysis (Ba, Be, Ca, Cu, Cr, K, Mg, Mn, Na, V, Fe, Co, Ni, Cu, Zn, Sr, Ti, Cd, and P) of pistachios samples was performed using inductively coupled plasma atomic emission spectrometry. Analysis of inorganic anions and organic acids (selenite, bromate, fumarate, malate, selenate, pyruvate, acetate, phosphate, and ascorbate) of pistachio samples was performed using capillary electrophoresis. Bulk carbon and nitrogen isotope ratios were performed using stable isotope MS. There were nearly 400 pistachio samples analyzed from the three major pistachio growing regions: Turkey, Iran, and California (United States). A computational evaluation of the trace element data sets was carried out using statistical pattern recognition methods including principal component analysis, canonical discriminant analysis, discriminant analysis, and neural network modeling. Several linear discriminant function models classified the data sets with 95% or higher accuracy. We report the development of a method combining elemental analysis and classification techniques that may be widely applied to the determination of the geographical origin of foods.