Detection of soybean oil adulteration in cow ghee (clarified milk fat): an ultrafast study using flash gas chromatography electronic nose coupled with multivariate chemometrics.

BACKGROUND: Cow ghee is one of the expensive edible fats in the dairy sector. Ghee is often adulterated with low-priced edible oils, like soybean oil, owing to its high market demand. The existing adulteration detection methods are time-consuming, requiring sample preparation and expertise in these fields. The possibility of detecting soybean oil adulteration (from 10% to 100%) in pure cow ghee was investigated in this study. The fingerprint information of volatile compounds was collected using a flash gas chromatography electronic nose (FGCEN) instrument. The classification results were studied using the pattern recognition chemometric models principal component analysis (PCA), soft independent modelling of class analogy (SIMCA), and discriminant function analysis (DFA). RESULTS: The most powerful fingerprint odor of all the samples identified from FGCEN analysis was acetaldehyde (Z)-4-heptenal, 2-propanol, ethyl propanoate, and pentan-2-one. The odor analysis investigation was accomplished with an average analysis time of 90 s. A clear differentiation of all the samples with an excellent classification accuracy of more than 99% was achieved with the PCA and DFA chemometric methods. However, the results of the SIMCA model showed that SIMCA could only be used to detect ghee adulteration at higher concentration levels (30% to 100%). The validation study shows good agreement between FGCEN and gas chromatography-mass spectrometry methods. CONCLUSION: The methodology demonstrated coupled with PCA and DFA methods for adulteration detection in ghee using FGCEN apparatus has been an efficient and convenient technique. This study explored the capability of the FGCEN instrument to tackle the adulteration problems in ghee. © 2022 Society of Chemical Industry.

Ghee Butter from Bovine Colostrum Reduces Inflammation in the Mouse Model of Acute Pancreatitis with Potential Involvement of Free Fatty Acid Receptors.

Acute pancreatitis (AP) is an inflammatory disease that causes severe tissue damage. Ghee butter from bovine colostrum (GBBC) is a clarified butter produced by heating milk fat to 40 °C and separating the precipitating protein. As colostrum mainly contains fatty acids (FAs), immunoglobulins, maternal immune cells, and cytokines, we hypothesized that it may exert anti-inflammatory effects. We investigated the effects of GBBC on experimental AP in mice. Two intraperitoneal (ip) injections of L-arginine (8%) were given 1 h apart to generate the AP murine model. After 12 h from the first L-arginine injection, mice were divided into the following experimental groups: AP mice treated with GBBC (oral gavage (po) every 12 h) and non-treated AP mice (po vehicle every 12 h). Control animals received vehicle only. At 72 h, mice were euthanized. Histopathological examination along with myeloperoxidase (MPO) and amylase/lipase activity assays were performed. In a separate set of experiments, FFAR1 and FFAR4 antagonists were used to verify the involvement of respective receptors. Administration of GBBC decreased MPO activity in the pancreas and lungs along with the microscopical severity of AP in mice. Moreover, treatment with GBBC normalized pancreatic enzyme activity. FFAR1 and FFAR4 antagonists tended to reverse the anti-inflammatory effect of GBBC in mouse AP. Our results suggest that GBBC displays anti-inflammatory effects in the mouse model of AP, with the putative involvement of FFARs. This is the first study to show the anti-inflammatory potential of a nutritional supplement derived from GBBC.

Fluorescence Spectroscopy Based Detection of Adulteration in Desi Ghee.

Desi ghee, obtained by buffalo and cow milk, is highly expensive because it contains valuable vitamins and conjugated linoleic acid (CLA). Its high demand and cost result in to its adulteration with inferior banaspati ghee. In this study, Fluorescence spectroscopy along with multivariate analysis has been utilised for the detection and quantification of adulteration. Spectroscopic analysis showed that buffalo ghee contains more vitamins and CLA than cow, whereas cow ghee is enriched with beta-carotene. For multivariate analysis, principle component analysis (PCA) and partial least square regression (PLSR) have been applied on the spectral data for the determination of adulteration. PLSR model was authenticated by predicting 23 unknown samples including 3 commercial brands of desi ghee. The root mean square error in prediction (RMSEP) of unknown samples was found to be 1.7 which is a reasonable value for quantitative prediction. Due to non-destructive and requiring no sample pre-treatment, this method can effectively be employed as on line characterization tool for the food safety assurance.

Characterization of Desi Ghee Extracted by Different Methods Using Fluorescence Spectroscopy.

In the current study, the effect of ghee extraction methods (direct cream DC, milk butter MB and milk skin MS) on its molecular composition has been investigated using Fluorescence spectroscopy. The excitation wavelength of 300 nm was found the best to produce pronounced spectral signatures of beta-carotene, vitamins and conjugated linoleic acid (CLA) in both cow and buffalo ghee types. Principal component analysis (PCA) has been applied on the spectral data to visualize the classification among ghee samples extracted by three methods. Both cow and buffalo ghee contain spectral signatures of vitamin A, E, K, D and CLA which has been verified through plotting loading vectors. The analysis of loading plots has been suggested that for cow ghee, MS extraction method conserve relatively higher concentration of beta carotene while DC and MB methods are a good choice for preserving relatively more concentrations of vitamins D, E and K. Similarly, for buffalo ghee, MS extraction method appear with higher concentration of CLA, whereas DC extraction method looks to preserve relatively higher concentration of vitamin A while MB method retains relatively low concentration of CLA and vitamins as compared to other two methods.

Cow ghee fortified ocular topical microemulsion; in vitro, ex vivo, and in vivo evaluation.

Aim: Utility of cow ghee (CG) as permeation enhancer in development of topical ocular microemulsion (ME) for delivery of fluocinolone acetonide (FA) to posterior eye. Methods: For ME preparation, oil, surfactant and cosurfactant were screened based on solubility of FA. Pseudoternary phase diagrams were constructed to determine their ratios. The developed MEs were characterised for their physicochemical properties like size, polydispersity index, zeta potential, and stability etc. They were evaluated for ex vivo permeation and irritation. In vivo pharmacokinetic studies were performed on Sprague dawley rats. Results: Lauroglycol as oil, labrasol as surfactant and Transcutol as cosurfactant were selected. The optimised ratio of oil:surfactant:cosurfactant:water was 4:23:23:50. The developed FA loaded ME fortified with CG was characterised. Ex vivo study revealed higher permeation and non-irritancy. In vivo pharmacokinetic study showed retention of CG fortified ME in posterior rat eye. Conclusion: Present investigation established CG as permeation enhancer for ocular topical formulation.

Traditional Tibetan Ghee: Physicochemical Characteristics and Fatty Acid Composition.

Fifty traditional Tibetan ghee (TTG) varieties were collected and analyzed their systematic characteristic indices, including physicochemical parameters, minerals, fatty acid composition, and thermal behavior. Results show that TTG contains a large amount of fat (71.68%-93.3%) and a small quantity of protein (0.51%-1.81%). The acid and peroxide values of TTG vary from 0.02 to 1.30 mg/g and 0.07 to 5.93 meq/kg, respectively. The content of minerals varied with altitude level and region significantly (p < 0.05), and the regional variations of fatty acids in TTG were also observed significantly, these differences may be due to the high unsaturated fatty acids level in the cow diets.

Characterization of desi ghee obtained from different extraction methods using Raman spectroscopy.

In this study, the potential of Raman spectroscopy has been utilized to characterize the methods direct cream (DC), milk butter (MB) and milk skin (MS) used for the extraction of desi ghee from buffalo and cow milk. Raman spectra from six types of ghee samples extracted by above methods were acquired using two laser wavelengths of 532 and 785 nm. The Raman spectra of cow ghee revealed that it contains three bands of beta-carotene at 1005, 1156 and 1520 cm-1 which differentiated it from buffalo ghee. To highlight small spectral differences, statistical analysis through principal component analysis (PCA) has been performed on the Raman spectra of ghee samples to reach subsequent conclusion. Based on the variations in molecular composition of cow ghee samples, it has been found that DC method retain relatively higher concentration of beta-carotene and MB method contain higher concentration of conjugated linoleic acid (CLA) and fatty acids than MS method. Similarly, DC & MS methods were found best for retaining relatively higher concentration of CLA and fatty acids in buffalo ghee as compared to MB method which retains relatively higher concentration of fatty acids.

Rapid and noninvasive quality control of anhydrous milk fat by PTR-MS: The effect of storage time and packaging.

In this study, proton transfer reaction-mass spectrometry (PTR-MS), coupled with a time-of-flight mass analyzer and a multipurpose automatic sampler, was evaluated as a rapid and nondestructive tool for the quality control of anhydrous milk fat. Anhydrous milk fats packed in cardboard and bag-in-box were compared during refrigerated shelf life at 4°C for 9 months. Anhydrous milk fat samples were taken at 120, 180, and 240 days and measured by PTR-MS during storage at 50°C for 11 days. Univariate and multivariate data analysis were performed in order to classify samples according to the packaging type and compare aromatic profiles. Markers related to both packaging and storage duration were identified, and all stored samples were clearly distinguishable from reference fresh samples. Significant differences in some key butter aroma compounds such as 2-pentanone, 2-heptanone, 2/3-methylbutanal, acetoin, and butanoic acid were observed between different types of packaging. During the refrigerated storage, differences related to packaging are more evident, while during the storage at 50°C, the fat oxidation induced by the high temperature becomes the most relevant phenomenon independently of the packaging type. These results indicate the importance of avoiding anhydrous milk fat storage at 50°C for long times during industrial production processes. All together data demonstrated the viability of PTR-MS as a rapid and high-sensitivity tool in agroindustry quality control program.

Studying heating effects on desi ghee obtained from buffalo milk using fluorescence spectroscopy.

Characterisation and thermal deterioration of desi ghee obtained from buffalo milk is presented for the first time using the potential of Fluorescence spectroscopy. The emission bands in non-heated desi ghee centred at 375 nm is labelled for vitamin D, 390 nm for vitamin K, 440-460 nm for isomers of conjugated linoleic acid (CLA), 490 nm for vitamin A and the region 620-700 nm is assigned to chlorophyll contents. Fluorescence emission spectra from all the heated and non-heated ghee samples were recorded using excitation wavelengths at 280, and 410 nm which were found best for getting maximum spectral signatures. Heating of desi ghee affects its molecular composition, however, the temperature range from 140 to 170°C may be defined safe for cooking /frying where it does not lose much of its molecular composition. Further, the rise in temperature induces prominent spectral variations which confirm the deterioration of valuable vitamins, isomers of CLA and chlorophyll contents. Fluorescence emission peak at 552 nm shows oxidation product and an increase in its intensity with the rise in temperature is observed. In order to classify heated samples at different temperatures, principal component analysis (PCA) has been applied on heated and non-heated ghee samples that further elucidated the temperature effects.

Heating Effects of Desi Ghee Using Raman Spectroscopy.

Raman spectroscopy as a fast and nondestructive technique has been used to investigate heating effects on Desi ghee during frying/cooking of food for the first time. A temperature in the range of 140-180℃ has been investigated within which Desi ghee can be used safely for cooking/frying without much alteration of its natural molecular composition. In addition, heating effects in case of reuse, heating for different times, and cooking inside pressure cookers are also presented. An excitation laser at 785 nm has been used to obtain Raman spectra and the range of 540-1800 cm-1 is found to contain prominent spectral bands. Prominent variations have been observed in the Raman bands of 560-770 cm-1, 790-1160 cm-1, and 1180-1285 cm-1 with the rise in temperature. The spectral variations have been verified using classifier principal component analysis. It has been found that Desi ghee can be reused if heated below 180℃ and it can be heated up to 30 min without any appreciable molecular changes if a controlled heating can be managed.

Robust new NIRS coupled with multivariate methods for the detection and quantification of tallow adulteration in clarified butter samples.

Cows' butterfat may be adulterated with animal fat materials like tallow which causes increased serum cholesterol and triglycerides levels upon consumption. There is no reliable technique to detect and quantify tallow adulteration in butter samples in a feasible way. In this study a highly sensitive near-infrared (NIR) spectroscopy combined with chemometric methods was developed to detect as well as quantify the level of tallow adulterant in clarified butter samples. For this investigation the pure clarified butter samples were intentionally adulterated with tallow at the following percentage levels: 1%, 3%, 5%, 7%, 9%, 11%, 13%, 15%, 17% and 20% (wt/wt). Altogether 99 clarified butter samples were used including nine pure samples (un-adulterated clarified butter) and 90 clarified butter samples adulterated with tallow. Each sample was analysed by using NIR spectroscopy in the reflection mode in the range 10,000-4000 cm-1, at 2 cm-1 resolution and using the transflectance sample accessory which provided a total path length of 0.5 mm. Chemometric models including principal components analysis (PCA), partial least-squares discriminant analysis (PLSDA), and partial least-squares regressions (PLSR) were applied for statistical treatment of the obtained NIR spectral data. The PLSDA model was employed to differentiate pure butter samples from those adulterated with tallow. The employed model was then externally cross-validated by using a test set which included 30% of the total butter samples. The excellent performance of the model was proved by the low RMSEP value of 1.537% and the high correlation factor of 0.95. This newly developed method is robust, non-destructive, highly sensitive, and economical with very minor sample preparation and good ability to quantify less than 1.5% of tallow adulteration in clarified butter samples.

Occurrence and spatial distribution of pesticide residues in butter and ghee (clarified butter fat) in Punjab (India).

The present study was undertaken to monitor organochlorine, organophosphate, and synthetic pyrethroid pesticide residues in butter (n = 55) and ghee (n = 56) samples collected from three different regions of Punjab. The estimation of pesticide residues was done by multiple residue analytical technique using gas chromatography equipped with GC-ECD and GC-FTD. The confirmation of residues was done on gas chromatography mass spectrometry in both selective ion monitoring (SIM) and scan mode. Results indicated the presence of hexacholorocyclohexane (HCH) and p,p' DDE as predominant contaminant in both butter and ghee. Residues of HCH were detected in 25 and 23% samples of butter and ghee, respectively, while residues of p,p' DDE were recorded in 29 and 25% of butter and ghee samples, respectively. None of the butter and ghee sample violated the MRL values of 200 ng g(-1) for HCH and 1250 ng g(-1) for dichorodiphenyl tricholorethane (DDT). The presence of endosulfan, cypermethrin, fenvalerate, deltamethrin, and chlorpyrifos were observed in a few butter and ghee samples at traces. The spatial variation for comparative occurrence of pesticide residues indicated higher levels in the south-western region of Punjab. Additionally, the temporal variation indicated the significant reduction of HCH and DDT levels in butter and ghee in Punjab.

Comparison of five analytical methods for the determination of peroxide value in oxidized ghee.

In the present study, a comparison of five peroxide analytical methods was performed using oxidized ghee. The methods included the three iodometric titration viz. Bureau of Indian Standard (BIS), Association of Analytical Communities (AOAC) and American Oil Chemists' Society (AOCS), and two colorimetric methods, the ferrous xylenol orange (FOX) and ferric thiocyanate (International Dairy Federation, IDF) methods based on oxidation of iron. Six ghee samples were stored at 80 °C to accelerate deterioration and sampled periodically (every 48 h) for peroxides. Results were compared using the five methods for analysis as well as a flavor score (9 point hedonic scale). The correlation coefficients obtained using the different methods were in the order: FOX (-0.836) > IDF (-0.821) > AOCS (-0.798) > AOAC (-0.795) > BIS (-0.754). Thus, among the five methods used for determination of peroxide value of ghee during storage, the highest coefficient of correlation was obtained for the FOX method. The high correlations between the FOX and flavor data indicated that FOX was the most suitable method tested to determine peroxide value in oxidized ghee.