• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1617-1617
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• 2001

It is well known now that near infrared spectroscopy (NIRS) is a fast, no destructive, and inexpensive analytical technique that could be used to classify, identify, and authenticate a wide range of foods and food items. Therefore, the main aims of this study were to provide a new insight into the authentication of five strawberry (Fragaria x ananassa) varieties and to correlate them with geographical zones and the propagating methods used. Three weeks plants of five different strawberry varieties (F. x ananassa Duch. cv Camarosa, Seascape, Chandler, F. Chiloensis, and F. Virginiana) were cultivated in vitro first and then transferred to pots with special soil, and grown in a greenhouse at CINVESTAV, all varieties were acquired from California (USA). After 18 months, ten leaves from each variety were collected. Transmission spectra from each leave were recorded over a range of 10, 000-4, 000 cm$-^{1}$, 32 scans of each strawberry leave were collected using a resolution of 4 cm$-^{1}$ with a Paragon IdentiCheck FT-NIR System Spectrometer. Triplicates of each strawberry leave were used. All spectra were analyzed using principal component analysis (PCA) and soft independent modeling class analogy (SIMCA). The optimum number of components to be used in the regression was automatically determined by the software. Camarosa was the only variety grown from the same shoot but propagated by a different method (direct or in vitro). Five different classes (varieties) or clusters were observed among samples, however, larger inter class distances were presented by the two wildtype samples (F. Chiloensis and F. Virginiana). Camarosa direct and Camarosa in vitro displayed a small overlapping region between them. On the other hand, Seascape variety presented the smallest rejection percentage among all varieties (more similarities with the rest of the samples). Therefore, it can be concluded that the application of NIRS technique allowed the authentication of all strawberry varieties and geographical origin as well. It was also possible to form subclasses of the same materials. The results presented here demonstrate that NIRS is a very powerful and promising analytical tool since all materials were authenticated and classified based on their variety, origin, and treatment. This is of a tremendous relevance since the variety and origin of a plant material can be established even before it gives its typical fruit or flower.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.4105-4105
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• 2001

Near infrared spectroscopy (NIRS) was employed to qualify and quantify on survival, the injury rate and apoptosis of the human breast cancer cell line MCF-7 cells. MCF-7 cells were cultured in RPMI medium supplemented with 10% FCS in a 95% air and 5% CO2 atmosphere at 37$^{\circ}C$. For the viable cells preparation, cells were de-touched by 0.1% of trypsin treatment and washed with RPMI supplemented with 10% FCS medium by centrifugation at 1000 rpm for 3min. For the dead cells preparation, cells were de-touched by a cell scraper. The cells were counted by a hemacytometer, and the viability was estimated by the exclusion method with frypan blue dye. Each viable and dead cells were suspended in PBS (phosphate bufferred saline) or milk at the cell density desired. For the quantitative determination of cell death by measuring the LDH (lactate dehydrogenase) activity liberated from cells with cell membrane injuries, LDH-Cytotoxic Test Wako (Wako, Pure Pharmaceutical Co. Ltd., Japan) was used. We found that NIRS measurement of MCF-7 cells at the density range could evaluate and monitor the different characteristics of living cells and dead cells. The spectral analysis was performed in two wavelength ranges and with 1,4, 10 mm pathlength. Different spectral data pretreatment and chemometrics methods were used. We applied SIMCA classificator on spectral data of living and dead cells and obtained good accuracy when identifying each class. Bigger variation in the spectra of living cells with different concentrations was observed when compared to the same concentrations of dead cells. PLS was used to measure the number of cells in PBS. The best model for measurement of dead cells, as well as living cells, was developed when raw spectra in the 600-1098 nm region and 4 mm pathlength were used. Smoothing and second derivative spectral data pretreatment gave worst results. The analysis of PLS loading explained this result with the scatter effect found in the raw spectra and increased with the number of cells. Calibration for cell count in the 1100-2500 nm region showed to be very inaccurate.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1520-1520
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• 2001

Food adulteration is a serious consumer fraud and a potentially dangerous practice. Regulatory authorities and food processors require a rapid, non-destructive test to accurately confirm authenticity in a range of food products and raw materials. Olive oil is prime target for adulteration either on the basis of the processing treatments used for its extraction (extra virgin vs virgin vs ordinary oil) or its geographical origin (e.g. Greek vs Italian vs Spanish). As part of an investigation into this problem, some preliminary work focused on the ability of near infrared spectroscopy to discriminate between virgin olive oils from separate regions of the Mediterranean i. e. Crete and the Peloponese. A total of 46 oils were collected: 18 originated in Crete and 28 in the Peloponese. Oils were stored in a temperature-controlled room at 2$0^{\circ}C$ prior to spectral collection at room temperature (15-18$^{\circ}C$). Samples (approximately 0.5$m\ell$) were placed in the centre of the quartz window in a camlock reflectance cell; the gold-plated baking plate was then gently placed into the cell against the glass so as to minimize the formation of air bubbles. The rear of the camlock cell was then screwed into place producing a sample thickness of 0.5mm. Spectra were recorded between 400 and 2498nm at 2nm intervals on a NIR Systems 6500 scanning monochromator. Spectral collection took place over 2-3 days. Data were analysed using both WINISI and The Unscrambler software to investigate the possibility of discriminating between the oils from Crete and the Peloponese. A number of data pre-treatments were used and discriminant models were developed using discriminant PLS (WINISI & Unscrambler) and SIMCA (Unscrambler). Despite the small number of samples involved, a satisfactory discrimination between these two oil types was achieved. Graphical examination of principal component scores for each oil type also holds out the possibility of separating oils from either Crete and the Peloponese on the basis of districts within each region. These preliminary data suggest the potential of near infrared spectroscopy to act as a screening technique for the confirmation of geographic origin of extra virgin olive oils. The sample presentation strategy adopted uses only small volumes of material and produces high quality spectra.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1031-1031
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• 2001

The mammary gland is made up of remarkably sensitive tissue, which has the capability of producing a large volume of secretion, milk, under normal or healthy conditions. When bacteria enter the gland and establish an infection (mastitis), inflammation is initiated accompanied by an influx of white cells from the blood stream, by altered secretory function, and changes in the volume and composition of secretion. Cell numbers in milk are closely associated with inflammation and udder health. These somatic cell counts (SCC) are accepted as the international standard measurement of milk quality in dairy and for mastitis diagnosis. NIR Spectra of unhomogenized composite milk samples from 14 cows (healthy and mastitic), 7days after parturition and during the next 30 days of lactation were measured. Different multivariate analysis techniques were used to diagnose the disease at very early stage and determine how the spectral properties of milk vary with its composition and animal health. PLS model for prediction of somatic cell count (SCC) based on NIR milk spectra was made. The best accuracy of determination for the 1100-2500nm range was found using smoothed absorbance data and 10 PLS factors. The standard error of prediction for independent validation set of samples was 0.382, correlation coefficient 0.854 and the variation coefficient 7.63%. It has been found that SCC determination by NIR milk spectra was indirect and based on the related changes in milk composition. From the spectral changes, we learned that when mastitis occurred, the most significant factors that simultaneously influenced milk spectra were alteration of milk proteins and changes in ionic concentration of milk. It was consistent with the results we obtained further when applied 2DCOS. Two-dimensional correlation analysis of NIR milk spectra was done to assess the changes in milk composition, which occur when somatic cell count (SCC) levels vary. The synchronous correlation map revealed that when SCC increases, protein levels increase while water and lactose levels decrease. Results from the analysis of the asynchronous plot indicated that changes in water and fat absorptions occur before other milk components. In addition, the technique was used to assess the changes in milk during a period when SCC levels do not vary appreciably. Results indicated that milk components are in equilibrium and no appreciable change in a given component was seen with respect to another. This was found in both healthy and mastitic animals. However, milk components were found to vary with SCC content regardless of the range considered. This important finding demonstrates that 2-D correlation analysis may be used to track even subtle changes in milk composition in individual cows. To find out the right threshold for SCC when used for mastitis diagnosis at cow level, classification of milk samples was performed using soft independent modeling of class analogy (SIMCA) and different spectral data pretreatment. Two levels of SCC - 200 000 cells/$m\ell$ and 300 000 cells/$m\ell$, respectively, were set up and compared as thresholds to discriminate between healthy and mastitic cows. The best detection accuracy was found with 200 000 cells/$m\ell$ as threshold for mastitis and smoothed absorbance data: - 98% of the milk samples in the calibration set and 87% of the samples in the independent test set were correctly classified. When the spectral information was studied it was found that the successful mastitis diagnosis was based on reviling the spectral changes related to the corresponding changes in milk composition. NIRS combined with different ways of spectral data ruining can provide faster and nondestructive alternative to current methods for mastitis diagnosis and a new inside into disease understanding at molecular level.