Characterization and identification of surface crystals on smear-ripened cheese by polarized light microscopy.

Surface crystallization and radial demineralization of Ca, P, and Mg occur in smear-ripened cheese. Furthermore, crystals of ikaite, struvite, calcite, and brushite have been identified in cheese smears by powder X-ray diffractometry (PXRD), and ikaite and struvite exist in smears as single crystals. Polarized light microscopy (PLM) is a simple, inexpensive, and well-established method in geology to detect and identify single crystals. However, use of PLM to identify cheese crystals has not been reported previously. The specific objectives of this research were (1) to identify crystals in cheese smears using selected PLM criteria; (2) to compare identification by PLM against PXRD; and (3) to develop and evaluate a novel treatment for smear material to improve crystal analyses by both PLM and PXRD. Duplicate wheels of 4 cheeses produced by different manufacturers were obtained from retail sources. Scrapings of surface smears were prepared and analyzed by PLM and PXRD by previously described methods. Crystals were categorized by PLM based on angle of extinction (AE), birefringence behavior under crossed polarizers and quartz filters, and size and shape (circularity) by image analysis. Crystals observed by PLM fell almost exclusively into 2 readily differentiated groups based on birefringence behavior and estimated angle of extinction. Group 1 (n = 18) were highly birefringent with AE = 88-92°, whereas group 2 (n = 28) had no birefringence with AE = 13-26°. Group 2 crystals were significantly larger and more circular than group 1 crystals. Group 1 and 2 were identified as struvite and ikaite, respectively, based on known birefringence and AE characteristics. Struvite was identified in all 4 cheeses by PLM but in only 3 cheeses by PXRD. Ikaite was identified in 3 cheeses by PLM but in only 2 cheeses by PXRD. These discrepancies occurred because the smear scrapings from 1 cheese contained excessive amorphous matter that caused extreme background noise, potentially obscuring diffractogram peaks that may have been present. To minimize noise, smear scrapings were dispersed in aqueous NaOH (pH 10) before analyses, which resulted in consistent results by PXRD and PLM. The method also rendered high-quality images by PLM. Data suggest that PLM may offer a simple and inexpensive means to identify struvite, ikaite, and possibly other single crystals in cheese smears.

Powder X-ray diffraction can differentiate between enantiomeric variants of calcium lactate pentahydrate crystal in cheese.

Powder X-ray diffraction has been used for decades to identify crystals of calcium lactate pentahydrate in Cheddar cheese. According to this method, diffraction patterns are generated from a powdered sample of the crystals and compared with reference cards within a database that contains the diffraction patterns of known crystals. During a preliminary study of crystals harvested from various Cheddar cheese samples, we observed 2 slightly different but distinct diffraction patterns that suggested that calcium lactate pentahydrate may be present in 2 different crystalline forms. We hypothesized that the 2 diffraction patterns corresponded to 2 enantiomeric forms of calcium lactate pentahydrate (L- and DL-) that are believed to occur in Cheddar cheese, based on previous studies involving enzymatic analyses of the lactate enantiomers in crystals obtained from Cheddar cheeses. However, the powder X-ray diffraction database currently contains only one reference diffraction card under the title "calcium lactate pentahydrate." To resolve this apparent gap in the powder X-ray diffraction database, we generated diffraction patterns from reagent-grade calcium l-lactate pentahydrate and laboratory-synthesized calcium dl-lactate pentahydrate. From the resulting diffraction patterns we determined that the existing reference diffraction card corresponds to calcium dl-lactate pentahydrate and that the other form of calcium lactate pentahydrate observed in cheese crystals corresponds to calcium l-lactate pentahydrate. Therefore, this report presents detailed data from the 2 diffraction patterns, which may be used to prepare 2 reference diffraction cards that differentiate calcium l-lactate pentahydrate from calcium dl-lactate pentahydrate. Furthermore, we collected crystals from the exteriors and interiors of Cheddar cheeses to demonstrate the ability of powder X-ray diffraction to differentiate between the 2 forms of calcium lactate pentahydrate crystals in Cheddar cheeses. Powder X-ray diffraction results were validated using enzymatic assays for lactate enantiomers. These results demonstrated that powder X-ray diffraction can be used as a diagnostic tool to quickly identify different forms of calcium lactate pentahydrate that may occur in Cheddar cheese.

Crystal fingerprinting: elucidating the crystals of Cheddar, Parmigiano-Reggiano, Gouda, and soft washed-rind cheeses using powder x-ray diffractometry.

Crystals in cheese may be considered defects or positive features, depending on the variety and mode of production (industrial, artisanal). Powder x-ray diffractometry (PXRD) offers a simple means to identify and resolve complex combinations of crystals that contribute to cheese characteristics. The objective of the present research was to demonstrate the application of PXRD to study crystals from a range of different cheese types, specifically Cheddar, Parmigiano-Reggiano, Gouda, and soft washed-rind (smear ripened) cheeses. In studies of Parmigiano-Reggiano and long-aged Gouda, PXRD has confirmed that hard (crunchy) crystals that form abundantly within these cheeses consist of tyrosine. Furthermore, PXRD has tentatively identified the presence of an unusual form of crystalline leucine in large (up to 6 mm in diameter) spherical entities, or "pearls", that occur abundantly in 2-year-old Parmigiano Reggiano and long-aged Gouda cheeses, and on the surface of rindless hard Italian-type cheese. Ongoing investigations into the nature of these "pearls" are providing new insight into the roles that crystals play in the visual appearance and texture of long-aged cheeses. Crystals also sometimes develop profusely in the eyes of long-aged Gouda, which have been shown by PXRD to consist of tyrosine and the aforementioned presumptive form of crystalline leucine. Finally, crystals have been shown by PXRD to form in the smears of soft washed-rind cheeses. These crystals may be associated in some cheeses with gritty mouth feel and with zonal body softening that occurs during ripening. Heightened interest in artisanal cheeses highlights the need to better understand crystals and their contributions to cheese characteristics.