scholarly journals Effect of Fat on Protein Estimation in Milk and Its Correlation with Lactose in Different Milk Types: A Small-Scale Study

2020 ◽  
Author(s):  
Sweekruthi A. Shetty ◽  
Melissa F. Young ◽  
Sunita Taneja ◽  
Kannan Rangiah
Keyword(s):  
Sample Preparation ◽  
Human Milk ◽  
Internal Standard ◽  
Cow Milk ◽  
Selected Reaction Monitoring ◽  
Small Scale ◽  
Milk Samples ◽  
Protein Estimation ◽  
Animal Milk ◽  
Fat Removal

Background: Estimation of macronutrients like protein and lactose is important to assess the quality of milk. To estimate these two macronutrients, ten raw milk samples obtained from each group of different animals (cow, goat, buffalo), ten pasteurized cow milk and ten human milk samples were analysed. Methods: Bicinchoninic acid (BCA) method was used to estimate protein from different milk samples. Four different sample preparation protocols were compared to check the effect of fat on BCA based protein estimation: dilution (D), fat removal-protein precipitation (FR and PP), fat removal-dilution (FR and D) and dilution-fat removal (D and FR). For lactose quantification, ultrahigh-performance liquid chromatography-mass spectrometry-selected reaction monitoring (UHPLC-MS/SRM) method was developed and validated using 13C6 lactose as internal standard (ISTD).Result: Among these four different protocols, D and FR method showed consistent data for total protein content in animal milk (cow-3.16%, goat-3.21%, buffalo-3.81%, pasteurized-2.98%) and FR and PP showed consistent data in human milk samples (1.2%). Though BCA method is simple to use, proper sample preparation protocol has to be applied prior to protein estimation to avoid the interference due to fat or lactose. In case of lactose, inter-day validation showed the accuracy ranging from 97.13 to 100.54%, coefficient of variation varying between 0.1 to 1.53%, correlation R2=0.999. Lactose is in the range of 4.1 to 4.8% in animal milk and 6.6% in human milk samples. The internal ratio of lactose/protein (1.28 to 1.55 in animal milk and 5.33 in human milk) will be useful to differentiate human milk from animal milk type and to assess the milk quality.

scholarly journals Quantification of Mycotoxins in Animal Milk from India (FS14-04-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Rukshan Mehta ◽  
Sweekruthi Shetty ◽  
Melissa Fox Young ◽  
P Barry Ryan ◽  
Kannan Rangiah
Keyword(s):  
Tamil Nadu ◽  
Method Development ◽  
Matrix Effects ◽  
Southern India ◽  
Selected Reaction Monitoring ◽  
Indian States ◽  
Total N ◽  
Biologically Relevant ◽  
Milk Samples ◽  
Animal Milk

Abstract Objectives Animal milk can be contaminated with mycotoxins (secondary metabolites of fungi) through poor quality feed and may be a source of human exposure. Our objective was to develop and optimize a method to detect biologically relevant concentrations of 8 mycotoxins (Aflatoxins B1, B2, G1, G2, M1, M2; Ochratoxins A, B) in animal milk. Methods We used ultra-high performance liquid chromatography/tandem mass spectrometry using selected reaction monitoring (UHPLC/MS-SRM) to quantify mycotoxins in animal milk samples (total N = 38; n = 10 each from cow and commercial milk and n = 9 from buffalo and goat) from the southern Indian states of Karnataka and Tamil Nadu. Method development was conducted and stable isotope dilution employed, using AFB1-D3 for aflatoxins and OTA-D5 for ochratoxins. We validated the method and examined matrix effects, freeze-thaw and auto-sampler stability. Our dynamic ranges from quantification were between 7.8–5000 pg/mL. Results Among samples collected from Southern India, 8 of 10 cow [median 103.35 pg/mL; n = 3 > 500 pg/mL]; 0 of 9 buffalo and 10 of 10 commercial [median: 151.5 pg/mL], milk samples were above the LOQ. AFM2 was also seen in samples from both regions, but in lower quantities when compared to AFM1 [median (north): 25.8 pg/mL; median (south): 70.95 pg/mL]. All except 3 samples were below the LOQ (31.3 pg/mL) for OTA, however we detected a sodium adduct of OTA above LOQ, across samples. We found [Na-OTA] in goat milk [median: 5.9 ng/mL] > buffalo [median: 2.2 ng/mL] > commercial [median: 2.04 ng/mL] > cow [median: 0.8 ng/mL]. Other mycotoxins were seen in concentrations close to or below LOQ. We did not identify significant stability issues. Conclusions We developed a highly sensitive method with biologically relevant dynamic ranges for detection of mycotoxins in milk samples. We found AFM1, AFM2, and Na-OTA in milk samples from Southern India. Further studies with larger sample sizes are warranted to establish the extent of mycotoxin contamination in milk. Funding Sources Funded by University Research Committee, Emory University and International Society for Research in Human Milk and Lactation.

scholarly journals Triacylglycerol Analysis in Human Milk and Other Mammalian Species: Small-Scale Sample Preparation, Characterization, and Statistical Classification Using HPLC-ELSD Profiles

2015 ◽  
Vol 63 (24) ◽  
pp. 5761-5770 ◽  
Author(s):  
Isabel Ten-Doménech ◽  
Eduardo Beltrán-Iturat ◽  
José Manuel Herrero-Martínez ◽  
Juan Vicente Sancho-Llopis ◽  
Ernesto Francisco Simó-Alfonso
Keyword(s):  
Sample Preparation ◽  
Human Milk ◽  
Mammalian Species ◽  
Small Scale ◽  
Statistical Classification ◽  
Triacylglycerol Analysis

scholarly journals LC-MS/MS Determination of 21 Non-Steroidal Anti-Inflammatory Drugs Residues in Animal Milk and Muscles

Molecules ◽  
2021 ◽  
Vol 26 (19) ◽  
pp. 5892
Author(s):  
Marta Pietruk ◽  
Piotr Jedziniak ◽  
Małgorzata Olejnik
Keyword(s):  
Sample Preparation ◽  
European Commission ◽  
Coefficient Of Variation ◽  
Chromatographic Separation ◽  
Chromatographic Column ◽  
Commission Decision ◽  
Milk Samples ◽  
Animal Milk ◽  
Inflammatory Drugs

The presented procedure combines experience from two LC-MS/MS methods previously developed by our team for NSAIDs determination in meat and milk. The novelty was a modification of sample preparation and combining LC-MS/MS method for milk and muscle. The clean-up procedure was investigated, leading to a change from SPE to dSPE with C18 bulk sorbent. Unlike most of the existing methods, chromatographic separation was achieved on a C8 chromatographic column. This method was developed and validated under European Commission Decision 2002/657/EC. Recovery for milk samples values between 86.3% to 108%, with the coefficient of variation, varied from 5.51% to 16.2%. The recovery for muscle was calculated to be between 85.0% and 109%, and the coefficient of variation was—4.73% to 16.6%. The validation results prove that the method is suitable for confirmatory purposes in milk and muscle. Of 452 samples tested in 2019 and 2020, two have been identified as non-compliant.

scholarly journals Macro- and micro-element analysis in milk samples by inductively coupled plasma-optical emission spectrometry

2016 ◽  
pp. 51-62 ◽  
Author(s):  
Sanja Petrovic ◽  
Sasa Savic ◽  
Zivomir Petronijevic
Keyword(s):  
Human Milk ◽  
Inductively Coupled Plasma ◽  
Optical Emission ◽  
Fat Content ◽  
Cow Milk ◽  
Emission Spectrometry ◽  
Inductively Coupled ◽  
Milk Samples ◽  
Plasma Optical Emission Spectrometry ◽  
Total Fat

The paper describes the determination of Ag, Al, B, Ba, Bi, Ca, Cd, Co, Cr, Cu, Fe, Ga, In, K, Li, Mg, Mn, Na, Ni, Pb, Sr, Tl and Zn, as well as total fat content of milk samples, originated from different sources. The analyzed milk samples were: human milk, fresh cow milk, pasteurized cow milk from a local market, and reconstituted powder milk. The milk samples were obtained from Jablanica District (Serbia) territory. Preparation of samples for macro- and micro-analyses was done by wet digestion. Concentrations of the elements after digestion were determined by inductively coupled plasma optical emission spectrometry (ICP-OES). Total fat content of milk samples was determinate by the Weibull and Stoldt method. The results showed that potassium and calcium concentrations were the highest in all samples: 1840.64 - 2993.26 mg/L and 456.05 - 1318.08 mg/L, respectively. Of all heavy metals from the examined milk samples (copper, zinc, manganese, nickel, cadmium, and lead), the most common were zinc and copper, with approximately similar content in the range of 5 - 12 mg/l, while cadmium nickel and manganese were not detected at all. Samples of fresh cow milk and human milk showed the highest fat content of 3.6 and 4.2 %, respectively. Results for total fat and macro- and micro-analyses showed that fresh cow milk has the highest contents of fat and calcium, making it the most nutritious.

scholarly journals Microbial and Chemical Adulterants Assessment of Raw Cow Milk Collected from Dairy Farms of Hlabisa Villages, KwaZulu-Natal Province, South Africa

2019 ◽  
Author(s):  
N.H. Xulu ◽  
S. Jamal-Ally ◽  
K.D. Naidoo
Keyword(s):  
South Africa ◽  
Dairy Farms ◽  
The Body ◽  
Cow Milk ◽  
Small Scale ◽  
Total Bacterial Count ◽  
Dairy Farmers ◽  
Milk Samples ◽  
Kwazulu Natal ◽  
Small Scale Dairy

Background: Milk is one of the most nutritious foods providing a variety of proteins, fats, minerals, and vitamins needed to maintain, grow, and develop the body. The aim of this study was to assess microbial and chemical adulterants of raw cow milk collected from dairy farms of Hlabisa villages, KwaZulu-Natal Province, South Africa. Methods: A total of 68 raw cow milk samples were obtained from teats sampling points, milking buckets, and communal pooling buckets. The bacteriological analysis was conducted for the detection of various bacteria in milk samples. Biochemical tests were also done to detect some chemical adulterants in milk samples. Results: Total bacterial count of teats, milking buckets, and communal milk pooling buckets were 6.91, 6.06, and 6.06 log Colony Forming Unit (CFU)/ml, respectively. The most found chemical adulterant was urea detected in 23 out of 68 (33.8%) samples, followed by hydrogen peroxide showed in 22 out of 68 (32.3%) samples. However, none of the samples were contaminated with formalin, starch, and neutralizer. Conclusion: The present study revealed high microbial contamination of raw cow milk produced by rural small-scale dairy farmers of Hlabisa villages, KwaZulu-Natal Province, South Africa, indicating the lack of standard operating sanitation. It was also stated that raw milk samples contained various types of chemical adulterants that may lead to severe health problems. Good hygiene practices must be adopted by small-scale dairy farmers at every stage of their milk handling and processing.

Quantification of Vitamin D3 in Feed, Food, and Pharmaceuticals Using High-Performance Liquid Chromatography/Tandem Mass Spectrometry

2012 ◽  
Vol 95 (5) ◽  
pp. 1487-1494 ◽  
Author(s):  
Heiko S Schadt ◽  
Richard Gössl ◽  
Natalie Seibel ◽  
Claude-P Aebischer
Keyword(s):  
Sample Preparation ◽  
Electrospray Ionization ◽  
Vitamin D3 ◽  
High Performance ◽  
Certified Reference Materials ◽  
Direct Injection ◽  
Internal Standard ◽  
Selected Reaction Monitoring ◽  
Intermediate Precision ◽  
Liquid Chromatography Tandem Mass

Abstract A rapid, sensitive, and selective method for the quantification of vitamin D3 (cholecalciferol) in solid and liquid food, feed, and tablets based on HPLC/MS/MS has been developed and validated. The sample preparation procedure consists of a quick and robust alkaline saponification and liquid–liquid extraction, followed by direct injection of the organic extract into the HPLC/MS/MS system for analysis without any further concentration, reconstitution, or prepurification steps. The reduction in sample preparation time was achieved by applying a heart-cutting, two-dimensional chromatography technique prior to positive electrospray ionization selected reaction monitoring MS analysis. Total vitamin D3 (sum of previtamin D3 and vitamin D3) was quantified using an isotopically labeled internal standard. The ionization efficiency of previtamin D3 and vitamin D3 in the positive electrospray ionization mode was found to be very similar. The validation experiments included four feed matrixes, three types of tablets, and 12 food matrixes. The obtained recoveries were between 96.1 and 105.3%, and intermediate precision ranged from 1.32 to 15.6% RSD, with HorRat values between 0.07 and 0.65. For all samples, extraction efficiencies were above 95.8%. Analysis of two certified reference materials (SRM 1849 and BCR-122) gave accuracies of 102.4 and 99.8%, respectively.

Determination of Some Heavy Metals in Milk of Cow Grazing At Selected Areas, of Kaduna Metropolis

2017 ◽  
Vol 7 (7) ◽  
pp. 341 ◽  
Author(s):  
Mahmud Mohammed Imam ◽  
Zahra Muhammad ◽  
Amina Zakari
Keyword(s):  
Heavy Metals ◽  
Cadmium Toxicity ◽  
Research Work ◽  
The Body ◽  
Cow Milk ◽  
Scientific Model ◽  
Potential Health ◽  
Milk Samples ◽  
Lead And Cadmium ◽  
Digestion Technique

In this research work the concentration of zinc, copper, lead, chromium, cadmium, and nickel in cow milk samples obtained from four different grazing areas   (kakuri, kudendan, malali, kawo) of Kaduna metropolis. The samples were digested by wet digestion technique .The trace element were determined using bulk scientific model VPG 210 model  Atomic Absorption Spectrophotometer (AAS).. The concentration of the determined heavy metal were The result revealed that Cr,  Ni and Cd were not detected in milk samples from Kawo, Malali  and Kudendan whereas lead (Pb) is detected in all samples and found to be above  the stipulated limits of recommended dietary allowance (NRC,1989) given as 0.02mg/day. Cu and Zn are essential elements needed by the body for proper metabolism and as such their deficiency or excess is very dangerous for human health. However, they were found in all samples and are within the recommended limits while Cd (2.13 – 3.15 mg/kg) in milk samples from Kakuri was found to be above such limit (0.5mg/day). Cow milk samples analyzed for heavy metals in this research work pose a threat of lead and cadmium toxicity due to their exposure to direct sources of air, water and plants in these grazing areas, thereby, resulting to a potential health risk to the consumers.

Development and validation of a LC-MS/MS method for quantification of mobocertinib (TAK-788) in plasma and its application to pharmacokinetic study in rats

2020 ◽  
Vol 23 ◽  
Author(s):  
Bo Li ◽  
Jin Wang ◽  
Xinyao Dou ◽  
Xinjie Zhang ◽  
Xianbei Xue ◽  
...  
Keyword(s):  
Oral Administration ◽  
Pharmacokinetic Study ◽  
High Performance ◽  
Kinase Inhibitor ◽  
Internal Standard ◽  
Selected Reaction Monitoring ◽  
Precipitation Method ◽  
Plasma Samples ◽  
Bioanalytical Method Validation ◽  
Extraction Recovery

Aim and Objective:: An analytical method for the determination of mobocertinib, an investigational tyrosine kinase inhibitor, was developed and optimized by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) in rat plasma. Materials and Methods:: Plasma samples were pretreated by the protein precipitation method with a methanol solution of osimertinib as the internal standard (IS). Chromatographic separation was performed using an Inertsil ODS-3 column (50 mm × 4.6 mm, I.D. 5 μm) column with the temperature maintained at 40 °C. The mobile phase consisted of water (containing 0.1% formic acid) and methanol in a gradient mode at a flow rate of 0.5 mL/min. Mass spectrometric detection was carried out in the selected reaction monitoring (SRM) mode with positive electrospray ionization, and the mass transitions of mobocertinib and osimertinib were m/z 587.01 → 71.88 and m/z 499.80 → 71.94, respectively. The method was validated in terms of selectivity, linearity, accuracy and precision, extraction recovery and matrix effect, stability and carryover as per the guidelines for bioanalytical method validation (FDA, 2018). The method was applied to the pharmacokinetic study of mobocertinib in rats by oral gavage at the doses of 2, 6, and 18 mg/kg. A total of 216 plasma samples from 18 rats were analyzed. Results:: It showed good linearity over the range of 1-1000 ng/mL (R2 = 0.9957). The intra-batch accuracy was within 94.65-102.59% and the precision was within 5.49-10.46%. The inter-batch accuracy was within 97.08-102.25% with a precision of 7.54-10.13%. The extraction recovery and matrix factor were acceptable for the bioanalysis of mobocertinib. Additionally, mobocertinib was found to be stable under the detected conditions. Mobocertinib showed linear pharmacokinetic characteristics following oral administration to rats at 2.0-18.0 mg/kg. Conclusion:: The developed and validated method was successfully employed in the pharmacokinetic study in rats following oral administration of mobocertinib at the doses of 2, 6, and 18 mg/kg.

scholarly journals COVID-19 Vaccine mRNABNT162b2 Elicits Human Antibody Response in Milk of Breastfeeding Women

Vaccines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 785
Author(s):  
Maurizio Guida ◽  
Daniela Terracciano ◽  
Michele Cennamo ◽  
Federica Aiello ◽  
Evelina La Civita ◽  
...  
Keyword(s):  
Breast Milk ◽  
Antibody Response ◽  
Human Milk ◽  
Human Antibody ◽  
Serum Antibodies ◽  
Serum Samples ◽  
Low Concentration ◽  
Milk Samples ◽  
Milk Antibodies ◽  
Roche Diagnostics

Objective: The objective of this research is to demonstrate the release of SARS-CoV-2 Spike (S) antibodies in human milk samples obtained by patients who have been vaccinated with mRNABNT162b2 vaccine. Methods: Milk and serum samples were collected in 10 volunteers 20 days after the first dose and 7 seven days after the second dose of the mRNABNT162b2 vaccine. Anti-SARS-CoV-2 S antibodies were measured by the Elecsys® Anti-SARS-CoV-2 S ECLIA assay (Roche Diagnostics AG, Rotkreuz, Switzerland), a quantitative electrochemiluminescence immunometric method. Results: At first sample, anti-SARS-CoV-2 S antibodies were detected in all serum samples (103.9 ± 54.9 U/mL) and only in two (40%) milk samples with a low concentration (1.2 ± 0.3 U/mL). At the second sample, collected 7 days after the second dose, anti-SARS-CoV-2 S antibodies were detected in all serum samples (3875.7 ± 3504.6 UI/mL) and in all milk samples (41.5 ± 47.5 UI/mL). No correlation was found between the level of serum and milk antibodies; the milk antibodies/serum antibodies ratio was on average 2% (range: 0.2–8.4%). Conclusion: We demonstrated a release of anti-SARS-CoV-2 S antibodies in the breast milk of women vaccinated with mRNABNT162b2. Vaccinating breastfeeding women could be a strategy to protect their infants from COVID-19 infection.

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