New Analytical Tools for Unmasking Frauds in Raw Milk-Based Dairy Products: Assessment, Validation and Application to Fiore Sardo PDO Cheese of a RP-HPLC Method for the Evaluation of the α-l-Fucosidase Activity

The activity of α-l-fucosidase (FSC) has been measured for the first time in Fiore Sardo PDO (Protected Designation of Origin) raw milk cheese. To do this, a RP-HPLC method has been developed, validated and tested on a reliable sampling of cheese experimentally produced in laboratory batches. Three experimental factors have been considered in this work: the thermal treatment undergone by the milk, the lactation period and the ripening time of cheese. Results obtained have evidenced: (i) a meaningful reduction in the activity of FSC from cheeses produced using raw milk to those obtained by thermized milk; (ii) an increase in the activity of FSC during the first months of lactation period (from December to February), followed by a substantial constancy in the central and final months of lactation (from February to May); (iii) the enzyme activity is independent of the ripening time. This method might be useful in revealing frauds related to the use of mild thermal treatments of the milk when this is not allowed as for Fiore Sardo PDO cheese but also for several PDO cheeses for which the specifications establish that raw milk only must be used for their production.

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Biogenic Amines in Traditional Fiore Sardo PDO Sheep Cheese: Assessment, Validation and Application of an RP-HPLC-DAD-UV Method

Separations ◽

10.3390/separations6010011 ◽

2019 ◽

Vol 6 (1) ◽

pp. 11 ◽

Cited By ~ 1

Author(s):

Claudia Zazzu ◽

Margherita Addis ◽

Marco Caredda ◽

Maria Francesca Scintu ◽

Giovanni Piredda ◽

...

Keyword(s):

Biogenic Amines ◽

Total Concentration ◽

Great Variability ◽

Quantitative Distribution ◽

Qualitative And Quantitative ◽

Rp Hplc ◽

Protected Designation Of Origin ◽

Assessment Validation ◽

The Many ◽

First Time

This contribution aimed to measure for the first time the amount of biogenic amines (BAs) in one of the most ancient and traditional sheep cheese produced in Sardinia, Italy: the Protected Designation of Origin (PDO) Fiore Sardo. To achieve this, an original RP-HPLC-DAD-UV method has been developed that was completely validated in terms of LoD, LoQ, linearity, precision and trueness, and tested on 36 real Fiore Sardo PDO cheese samples produced by four different cheesemakers and marketed by four stores. The average total concentration of the eight BAs (i.e., tyramine, tryptamine, histidine, putrescine, cadaverine, 2-phenylethylamine, spermine and spermidine) measured in Fiore Sardo cheese was 700 mg/kg, with a range between 170 mg/kg and 1,100 mg/kg. A great variability in the total amount of BAs has been evidenced among the Fiore Sardo marketed in the four stores as well as for the cheeses purchased in different times in the same store. Tyramine (350 mg/kg), putrescine (150 mg/kg), histamine (80 mg/kg) and cadaverine (30 mg/kg) are the most abundant BAs found in this matrix. Among the many factors concurring, the dominant microflora of Fiore Sardo PDO is likely the principal cause of the qualitative and quantitative distribution of BAs in this matrix. Finally, the total amount of BAs found in Fiore Sardo PDO is not able to cause any health alert situation for consumers.

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Biogenic Amines in Traditional Fiore Sardo PDO Sheep Cheese: Assessment, Validation and Application of a RP-HPLC-DAD-UV Method

10.20944/preprints201901.0290.v1 ◽

2019 ◽

Author(s):

Claudia Zazzu ◽

Margherita Addis ◽

Marco Caredda ◽

Maria Francesca Scintu ◽

Giovanni Piredda ◽

...

Keyword(s):

Biogenic Amines ◽

Total Concentration ◽

Great Variability ◽

Quantitative Distribution ◽

Qualitative And Quantitative ◽

Rp Hplc ◽

Protected Designation Of Origin ◽

Assessment Validation ◽

First Time

This contribution is aimed to measure for the first time the amount of biogenic amines (BAs) in one of the most ancient and traditional sheep cheese produced in Sardinia, Italy: the Protected Designation of Origin (PDO) Fiore Sardo. For doing this, an original RP-HPLC-DAD-UV method has been developed, completely validated in terms of LoD, LoQ, linearity, precision and trueness, and tested on 36 real Fiore Sardo PDO cheese samples produced by four different cheesemakers and marketed by four stores. The average total concentration of the eight BAs (i.e. tyramine, tryptamine, histidine, putrescine, cadaverine, 2-phenylethylamine, spermine and spermidine) measured in Fiore Sardo cheese was 700 mg/kg, with a range between 170 mg/kg and 1100 mg/kg. A great variability in the total amount of BAs has been evidenced among the Fiore Sardo marketed in the four stores as well as for the cheeses purchased in different times in the same store. Tyramine (350 mg/kg), putrescine (150 mg/kg), histamine (80 mg/kg) and cadaverine (30 mg/kg) are the most abundant BAs found in this matrix. Among many factors concurring, the dominant microflora of Fiore Sardo PDO is likely the principal cause of the qualitative and quantitative distribution of BAs in this matrix. Finally, the total amount of BAs found in Fiore Sardo PDO is not able to cause any situation of health alert for consumers.

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Survival of Selected Pathogenic Bacteria during PDO Pecorino Romano Cheese Ripening

Dairy ◽

10.3390/dairy1030020 ◽

2020 ◽

Vol 1 (3) ◽

pp. 297-312

Author(s):

Giacomo Lai ◽

Rita Melillo ◽

Massimo Pes ◽

Margherita Addis ◽

Antonio Fadda ◽

...

Keyword(s):

Pathogenic Bacteria ◽

Salt Content ◽

Raw Milk ◽

Salmonella Spp ◽

Growth And Survival ◽

E Coli ◽

Cheese Ripening ◽

Protected Designation Of Origin ◽

Low Levels ◽

First Time

This study was conducted to assess, for the first time, the survival of the pathogenic bacteria Listeria monocytogenes, Salmonella spp., Escherichia coli O157:H7, and Staphylococcus aureus during the ripening of protected designation of origin (PDO) Pecorino Romano cheese. A total of twenty-four cheese-making trials (twelve from raw milk and twelve from thermized milk) were performed under the protocol specified by PDO requirements. Sheep cheese milk was first inoculated before processing with approximately 106 colony-forming unit (CFU) mL−1 of each considered pathogen and the experiment was repeated six times for each selected pathogen. Cheese composition and pathogens count were then evaluated in inoculated raw milk, thermized milk, and cheese after 1, 90, and 150 days of ripening. pH, moisture, water activity, and salt content of cheese were within the range of the commercial PDO Pecorino Romano cheese. All the cheeses made from raw and thermized milk were microbiologically safe after 90 days and 1 day from their production, respectively. In conclusion, when Pecorino Romano cheese is produced under PDO specifications, from raw or thermized milk, a combination of factors including the speed and extent of curd acidification in the first phase of the production, together with an intense salting and a long ripening time, preclude the possibility of growth and survival of L. monocytogenes, Salmonella spp., and E. coli O157:H7. Only S. aureus can be still detectable at such low levels that it does not pose a risk to consumers.

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Validated Chromatographic Methods for Simultaneous Estimation of Cinnarizine in Binary Mixture with Domperidone and Paracetamol in Tablets

Current Pharmaceutical Analysis ◽

10.2174/1573412914666180307154921 ◽

2019 ◽

Vol 15 (5) ◽

pp. 429-438

Author(s):

Marianne Alphonse Mahrouse ◽

Asmaa Ahmed El-Zaher ◽

Ahmed Mohammed Al-Ghani

Keyword(s):

Quality Control ◽

Hplc Method ◽

Simultaneous Estimation ◽

Chromatographic Methods ◽

Treatment And Prevention ◽

Rp Hplc ◽

Linear Relationships ◽

Ich Guidelines ◽

Analytical Tools

Background:Cinnarizine, an antihistaminic drug, is commonly formulated in combination with domperidone and with paracetamol for treatment and prevention of motion sickness and migraine.Objective:The aim of this work was to develop new, simple, precise and selective chromatographic methods (RP-HPLC and TLC-densitometric methods) for the determination of these drugs. These methods can be used as analytical tools in the routine examination in quality control laboratories.Methods:The first method was RP-HPLC method, the separation was carried out on an Inertsil® ODS- 3V C18 column (250 mm × 4.6 mm, 5 µm) using a mobile phase composed of methanol: acetonitrile (45: 55, v/v) at a flow rate of 1 ml/min. The detection was carried out at 220 nm. The second method was a TLC-densitometric method where the studied components were separated using a developing system composed of toluene: ethyl acetate: methanol: triethylamine (5: 4.3: 0.7: 0.5, v/v/v/v) on TLC silica gel 60 F254 plates, followed by densitometric scanning at 270 nm.Results:In RP-HPLC method, the peaks were sharp and well separated, the retention times were 5.25, 3.48 and 2.78 min, for cinnarizine, domperidone and paracetamol, respectively. Linearity was obtained over the concentration ranges 1-22, 0.75-16.5 and 25-550 µg/ml, for cinnarizine, domperidone and paracetamol, respectively. In TLC-densitometric method, good separation of spots and linear relationships were achieved over the concentration ranges of 0.2-2, 0.15-1.5 and 5-50 µg/spot, for cinnarizine, domperidone and paracetamol, respectively. Method validation was conducted according to ICH guidelines in terms of linearity, accuracy, selectivity, precision and robustness.Conclusion:The developed methods were applied for the determination of the cited drugs in tablets containing binary drug mixtures. The methods are simple and precise and can be used for routine analysis of the labelled drugs in combined dosage forms in quality control laboratories.

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Determination of Lactoferrin in Goat Milk by HPLC Method

Czech Journal of Food Sciences ◽

10.17221/944-cjfs ◽

2009 ◽

Vol 27 (Special Issue 1) ◽

pp. S102-S104 ◽

Cited By ~ 18

Author(s):

M. Dračková ◽

I. Borkovcová ◽

B. Janštová ◽

M. Naiserová ◽

H. Přidalová ◽

...

Keyword(s):

High Performance ◽

Goat Milk ◽

Raw Milk ◽

Hplc Method ◽

Average Concentration ◽

Array Detector ◽

Lactation Period ◽

The Czech Republic ◽

South Moravia

The aim of this study was the determination of lactoferrin in goat milk using HPLC method. Milk samples were collected at a goat farm in the South Moravia Region, the Czech Republic. It were established bulk tank samples of raw milk (n = 24) and pasteurised milk (nn = 27) that were collected during lactation. Lactoferrin contents were analysed by reverse phase high-performance liquid chromatography (RP-HPLC) with diode-array detector PDA 2996. Detection was carried out at the wavelength 205 nm. The average concentration of lactoferrin in goat milk was 120 ± 18 μg/ml. The lactoferrin content was increasing within the lactation period in the ranges of 98 ± 170 μg/ml in April to 149 ± 19 μg/ml in November. The heat treatment (pasteurisation at 72°C for 20 s) resulted in no significant effect on the lactoferrin content. No statistically significant differences (P = 0.05) were found between the values of raw and pasteurised goat milk.

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Assessment, Validation and Application to Real Samples of a RP-HPLC Method for the Determination of Guayulins A, B, C and D in Guayule Shrub

10.20944/preprints201802.0161.v2 ◽

2018 ◽

Author(s):

Nadia Spano ◽

Paola Meloni ◽

Ilenia Idda ◽

Alberto Mariani ◽

Maria Itria Pilo ◽

...

Keyword(s):

Sesquiterpene Lactones ◽

Biological Properties ◽

Hplc Method ◽

Raw Material ◽

Anticancer Activities ◽

Rp Hplc ◽

Assessment Validation ◽

Anisic Acid ◽

The Arid Regions

Guayule (Parthenium argentatum Gray) is a shrub native of the arid regions of Mexico. In the last decades, significant attention for its cultivation has risen because it is the raw material for the production of hypoallergenic natural rubber. Guayule biomass contains also high amounts of resin, which is not normally exploited in any way. Among other sesquiterpenic esters, guayulins (i.e. the parteniol esters of cinnamic acid, guayulin A, or of anisic acid, guayulin B) are contained in resin. In addition, minor amounts of guayulin C and guayulin D are formed by degradation/oxidation of guayulins A and B, respectively. Guayulins likely act as cinnamate and p-anisate reservoirs for Guayule shrub, in addition, it has been postulated that they might have a key role in the chemical defense system of Guayule. Furthermore, it seems reasonable that guayulins may possess significant biological properties (e.g. antibacterial and anticancer activities), in close analogy with those shown by sesquiterpene lactones contained in many other species of Parthenum genus. As a matter of fact, guayulins A and B play an important role in the synthesis of antineoplastics used in breast cancer treatment. In this contribution we propose an original and validated RP-HPLC approach to the simultaneous quantification of guayulins A, B, C and D. The procedure of resin extraction from Guayule biomass has been optimized in terms of both extraction method and solvent. RP-HPLC separation has been accomplished by an Ascentis® C18 column under isocratic elution with a 80:20 (v:v) acetonitrile:water mixture. Validation was carried out in terms of limits of detection and quantification, linearity, precision, and trueness. Finally, the method was tested with a number of fresh and seasoned samples of spontaneous Guayule shrub from Mexico.

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FACILE AND MANIFEST RPHPLC METHOD FOR THE DETERMINATION OF CAPTPRIL, LISIOPRIL, ENALPRIL AND DICLOFENAC SODIUM: ITS APPLICATIONS IN DOSAGE FORMULATIONS AND IN HUMAN SERUM

10.21065/19257430.40.4 ◽

2014 ◽

Vol 4 ◽

pp. 40

Author(s):

Safila Naveed ◽

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Human Serum ◽

High Performance ◽

Diclofenac Sodium ◽

Pharmaceutical Products ◽

Hplc Method ◽

Rp Hplc ◽

First Time

A simple, rapid, isocratic, high-performance liquid chromatography (RP-HPLC) method has been developed for the first time for simultaneous determination of ACE inhibitors (captopril, lisinopril and enalapril) and diclofenac sodium in bulk drugs, pharmaceutical products and human serum.

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Assessment, Validation and Application to Real Samples of an RP-HPLC Method for the Determination of Guayulins A, B, C and D in Guayule Shrub

Separations ◽

10.3390/separations5020023 ◽

2018 ◽

Vol 5 (2) ◽

pp. 23 ◽

Cited By ~ 2

Author(s):

Nadia Spano ◽

Paola Meloni ◽

Ilenia Idda ◽

Alberto Mariani ◽

Maria Pilo ◽

...

Keyword(s):

Hplc Method ◽

Real Samples ◽

Rp Hplc ◽

Assessment Validation

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Development and validation of HPLC and CE methods for simultaneous determination of amlodipine and atorvastatin in the presence of their acidic degradation products in tablets

Acta Pharmaceutica ◽

10.1515/acph-2016-0040 ◽

2016 ◽

Vol 66 (4) ◽

pp. 479-490 ◽

Cited By ~ 11

Author(s):

Said A. Hassan ◽

Eman S. Elzanfaly ◽

Salem Badr A. El-Zeany ◽

Maissa Y. Salem

Keyword(s):

Capillary Electrophoresis ◽

Degradation Products ◽

Hplc Method ◽

Rp Hplc ◽

Ich Guidelines ◽

Acidic Degradation ◽

Development And Validation ◽

First Time ◽

Hydrolysis Of

Abstract Two methods were developed for separation and quantitation of amlodipine (AML) and atorvastatin (ATV) in the presence of their acidic degradation products. The first method was a simple isocratic RP-HPLC method while the second was capillary electrophoresis (CE). Degradation products were obtained by acidic hydrolysis of the two drugs and their structures were elucidated for the first time by IR and MS spectra. Degradation products did not interfere with the determination of either drug and the assays were therefore stability-indicating. The linearity of the proposed methods was established over the ranges 1-50 μg mL-1 for AML and ATV in the HPLC method and in the range of 3-50 and 4-50 μg mL-1 for AML and ATV, respectively, in the CE method. The proposed methods were validated according to ICH guidelines. The methods were successfully applied to estimation of AML and ATV in combined tablets.

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Development and validation of a new analytical RP-HPLC method for simultaneous determination of Glibenclamide and Atenolol in bulk

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v10i3.1491 ◽

2019 ◽

Vol 10 (3) ◽

pp. 2433-2445

Author(s):

Anitha P ◽

Ramkanth S ◽

Satyanarayana S V

Keyword(s):

Hplc Method ◽

Fixed Dose ◽

Simultaneous Estimation ◽

Uv Detector ◽

Rp Hplc ◽

Ich Guidelines ◽

System Suitability ◽

Development And Validation ◽

First Time

A new, simple, reliable, fast, sensitive and economical RP-HPLC method was developed and validated for simultaneous estimation of two fixed-dose combinations frequently prescribed in coexisted chronic diseases such as diabetes (GLB) and hypertension (ATN) in bulk for the first time. The mobile phase used for the chromatographic runs consisted of 0.01N potassium dihydrogen ortho phosphate (pH 4.8) and acetonitrile (55:45, v/v). The separation was achieved on column (BDS C18 250 x 2.1mm, 1.6m) using isocratic mode. Drug peaks were well separated and were detected by a UV detector at 235.0 nm. The method was linear at the concentration range 2.5-15µg/ml for Glibenclamide (GLB) and 6.25-37.5µg/ml for Atenolol (ATN), respectively. The method has been validated according to ICH guidelines with respect to system suitability, specificity, precision, accuracy and robustness. The method was validated for system suitability, linearity, accuracy, precision, detection, quantification limits and robustness and was found it is acceptable in the range of 2.5–15 µg/ml for GLB and 6.25–37.5 µg/ml for ATN. The LOD and LOQ of GLB was found to be 0.48 µg/ml and 1.47µg/ml and for ATN was found to be 0.72µg/ml and 2.20 µg/ml, respectively. The method was applied to drug interaction studies of GLB with ATN to illustrate the scope and application of the methods to manage two different therapeutic classes of drugs, as they may co-administered in concurrent diseases.

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