Validated Identity Test Method for Ginkgo biloba NHPs Using DNA-Based Species-Specific Hydrolysis PCR Probe

2019 ◽  
Vol 102 (6) ◽  
pp. 1779-1786 ◽  
Author(s):  
Shanmughanandhan Dhivya ◽  
Subramanyam Ragupathy ◽  
Prasad Kesanakurti ◽  
Shanmughanandhan Jeevitha ◽  
Isabella Della Noce ◽  
...  
Keyword(s):  
Ginkgo Biloba ◽  
Raw Materials ◽  
Target Material ◽  
Test Method ◽  
Target Species ◽  
Herbal Products ◽  
Hydrolysis Probe ◽  
Identity Test ◽  
Probe Assay ◽  
Species Specific

Background: There is considerable risk of adulteration of Ginkgo biloba herbal products in the natural health product (NHP) industry. Authentication of G. biloba products is challenging because of the standard, complex, analytical chemistry methods that may be too costly and not appropriate for both raw and finished products. Objective: We sought to develop and validate an alternative method to authenticate G. biloba herbal dietary supplements, based on the use of a species-specific hydrolysis PCR probe assay. Methods: A species-specific hydrolysis probe assay was developed, validated, and evaluated for the performance of the assay in accurately identifying the species of interest using the following analytical validation criteria: (1) specificity (accuracy) in identifying the target species ingredient, while not identifying other nontarget species, (2) sensitivity in detecting the smallest amount of the target material, and (3) reliability (repeatability and reproducibility) in detecting the target species in raw materials on a real-time PCR platform. Results: The species-specific hydrolysis probe assay was successfully developed for raw materials of G. biloba. The specificity of the assay was 100% to the target species. Efficiency of the assay was observed to be 99%, and the reliability of the assay was 100% for the raw/starting materials. Conclusions and Highlights: The method developed in this study is simple, rapid, and easy for supplement manufacturers to perform in their laboratories to ensure that their G. biloba supplements are authentic.

Validated Identity Test Method for Ginkgo biloba NHPs Using DNA-Based Species-Specific Hydrolysis PCR Probe

2019 ◽  
Vol 102 (6) ◽  
pp. 1779-1786 ◽  
Author(s):  
Shanmughanandhan Dhivya ◽  
Subramanyam Ragupathy ◽  
Prasad Kesanakurti ◽  
Shanmughanandhan Jeevitha ◽  
Isabella Della Noce ◽  
...  
Keyword(s):  
Ginkgo Biloba ◽  
Raw Materials ◽  
Target Material ◽  
Test Method ◽  
Target Species ◽  
Herbal Products ◽  
Hydrolysis Probe ◽  
Identity Test ◽  
Probe Assay ◽  
Species Specific

Abstract Background: There is considerable risk of adulteration of Ginkgo biloba herbal products in the natural health product (NHP) industry. Authentication of G. biloba products is challenging because of the standard, complex, analytical chemistry methods that may be too costly and not appropriate for both raw and finished products. Objective: We sought to develop and validate an alternative method to authenticate G. biloba herbal dietary supplements, based on the use of a species-specific hydrolysis PCR probe assay. Methods: A species-specific hydrolysis probe assay was developed, validated, and evaluated for the performance of the assay in accurately identifying the species of interest using the following analytical validation criteria: (1) specificity (accuracy) in identifying the target species ingredient, while not identifying other nontarget species, (2) sensitivity in detecting the smallest amount of the target material, and (3) reliability (repeatability and reproducibility) in detecting the target species in raw materials on a real-time PCR platform. Results: The species-specific hydrolysis probe assay was successfully developed for raw materials of G. biloba. The specificity of the assay was 100% to the target species. Efficiency of the assay was observed to be 99%, and the reliability of the assay was 100% for the raw/starting materials. Conclusions and Highlights: The method developed in this study is simple, rapid, and easy for supplement manufacturers to perform in their laboratories to ensure that their G. biloba supplements are authentic.

Validation and optimization of qPCR method for identification of Actaea Racemosa (Black cohosh) NHPs

2020 ◽  
Author(s):  
Jeevitha Shanmughanandhan ◽  
Dhivya Shanmughanandhan ◽  
Subramanyam Ragupathy ◽  
Thomas A Henry ◽  
Steven G Newmaster
Keyword(s):  
Dietary Supplements ◽  
Raw Materials ◽  
Black Cohosh ◽  
Powder Materials ◽  
Target Species ◽  
Herbal Products ◽  
Hydrolysis Probe ◽  
Actaea Racemosa ◽  
Probe Assay ◽  
Species Specific

Abstract BACKGROUND Actaea racemosa (Black cohosh) herbal dietary supplements are commonly used to treat menopausal symptoms in women. However, there is a considerable risk of contamination of A. racemosa herbal products in the natural health product (NHP) industry, impacting potential efficacy. Authentication of A. racemosa products is challenging because of the standard, multi-part analytical chemistry methods that may be too costly and not appropriate for both raw and finished products. OBJECTIVE In this paper, we discuss about developing and validating quick alternative biotechnology methods to authenticate A. racemosa herbal dietary supplements, based on the use of a species-specific hydrolysis PCR probe assay. METHODS A qPCR based species-specific hydrolysis probe assay was designed, validated, and optimized for precisely identifying the species of interest using the following analytical validation criteria: (1) specificity (accuracy) in determining the target species ingredient, while not identifying other non-target species, (2) sensitivity in detecting the smallest amount of the target material, and (3) reliability (repeatability and reproducibility) in detecting the target species in raw materials on a real-time PCR platform. RESULTS The results show that the species-specific hydrolysis probe assay was successfully developed for the raw materials and powders of A. racemosa. The specificity of the test was 100% to the target species. The efficiency of the assay was observed to be 99%, and the reliability of the assay was 100% for the raw/starting and powder materials. CONCLUSION The method developed in this study can be used to authenticate and perform qualitative analysis of A. racemosa supplements.

Recommendations for Validation of Real-Time PCR Methods for Molecular Diagnostic Identification of Botanicals

2019 ◽  
Vol 102 (6) ◽  
pp. 1767-1773 ◽  
Author(s):  
Steven G Newmaster ◽  
Dhivya Shanmughanandhan ◽  
Prasad Kesanakurti ◽  
Hanan Shehata ◽  
Adam Faller ◽  
...  
Keyword(s):  
Real Time ◽  
Foodborne Pathogens ◽  
Real Time Pcr ◽  
Target Material ◽  
Test Method ◽  
Alternative Methods ◽  
Molecular Diagnostic ◽  
Data Systems ◽  
Target Species ◽  
Commercial Laboratory

Abstract Background: PCR methods are the most commonly used DNA-based identity tool in the commercial food, beverage, and natural health product markets. These methods are routinely used to identify foodborne pathogens and allergens in food. Proper validation methods for some sectors have been established, while there are none in other markets, such as botanicals. Results: A survey of the literature indicates that some validation criteria are not addressed when developing PCR tests for botanicals. Objective: We provide recommendations for qualitative real-time PCR methods for validating identity tests for botanical ingredients. Methods: These include common criteria that underpin the development and validation of rigorous tests, including (1) the aim of the validation test, (2) the applicability of different matrix variants, (3) specificity in identifying the target species ingredient, (4) sensitivity in detecting the smallest amount of the target material, (5) repeatability of methods, (6) reproducibility in detecting the target species in both raw and processed materials, (7) practicability of the test in a commercial laboratory, and (8) comparison with alternative methods. In addition, we recommend additional criteria, according to which the practicability of the test method is evaluated by transferring the method to a second laboratory and by comparison with alternative methods. Conclusions and Highlights: We hope that these recommendations encourage further publication on the validation of PCR methods for many botanical ingredients. These properly validated PCR methods can be developed on small, real-time biotechnology that can be placed directly into the supply chain ledger in support of highly transparent data systems that support QC from the farm to the fork of the consumer.

Recommendations for Validation of Real-Time PCR Methods for Molecular Diagnostic Identification of Botanicals

2019 ◽  
Vol 102 (6) ◽  
pp. 1767-1773 ◽  
Author(s):  
Steven G. Newmaster ◽  
Dhivya Shanmughanandhan ◽  
Prasad Kesanakurti ◽  
Hanan Shehata ◽  
Adam Faller ◽  
...  
Keyword(s):  
Real Time ◽  
Foodborne Pathogens ◽  
Real Time Pcr ◽  
Target Material ◽  
Test Method ◽  
Alternative Methods ◽  
Molecular Diagnostic ◽  
Data Systems ◽  
Target Species ◽  
Commercial Laboratory

Background: PCR methods are the most commonly used DNA-based identity tool in the commercial food, beverage, and natural health product markets. These methods are routinely used to identify foodborne pathogens and allergens in food. Proper validation methods for some sectors have been established, while there are none in other markets, such as botanicals. Results: A survey of the literature indicates that some validation criteria are not addressed when developing PCR tests for botanicals. Objective: We provide recommendations for qualitative real-time PCR methods for validating identity tests for botanical ingredients. Methods: These include common criteria that underpin the development and validation of rigorous tests, including (1) the aim of the validation test, (2) the applicability of different matrix variants, (3) specificity in identifying the target species ingredient, (4) sensitivity in detecting the smallest amount of the target material, (5) repeatability of methods, (6) reproducibility in detecting the target species in both raw and processed materials, (7) practicability of the test in a commercial laboratory, and (8) comparison with alternative methods. In addition, we recommend additional criteria, according to which the practicability of the test method is evaluated by transferring the method to a second laboratory and by comparison with alternative methods. Conclusions and Highlights: We hope that these recommendations encourage further publication on the validation of PCR methods for many botanical ingredients. These properly validated PCR methods can be developed on small, real-time biotechnology that can be placed directly into the supply chain ledger in support of highly transparent data systems that support QC from the farm to the fork of the consumer.

scholarly journals A Health Risk Assessment of Lead and Other Metals in Pharmaceutical Herbal Products and Dietary Supplements Containing Ginkgo biloba in the Mexico City Metropolitan Area

2021 ◽  
Vol 18 (16) ◽  
pp. 8285
Author(s):  
Patricia Rojas ◽  
Elizabeth Ruiz-Sánchez ◽  
Camilo Ríos ◽  
Ángel Ruiz-Chow ◽  
Aldo A. Reséndiz-Albor
Keyword(s):  
Health Risk ◽  
Dietary Supplements ◽  
Mexico City ◽  
Metropolitan Area ◽  
Ginkgo Biloba ◽  
Graphite Furnace ◽  
Human Health Risk ◽  
Daily Intake ◽  
Herbal Remedies ◽  
Herbal Products

The use of the medicinal plant Ginkgo biloba has increased worldwide. However, G. biloba is capable of assimilating both essential and toxic metals, and the ingestion of contaminated products can cause damage to health. The aim of this study was to investigate the safety of manganese (Mn), copper (Cu), lead (Pb), arsenic (As), and cadmium (Cd) in 26 items containing Ginkgo biloba (pharmaceutical herbal products, dietary supplements, and traditional herbal remedies) purchased in the metropolitan area of Mexico City. Metal analysis was performed using a graphite furnace atomic absorption spectrometer. All of the products were contaminated with Pb, 54% of them with As, and 81% with Cd. The lowest values of Pb, As, and Cd were detected in pharmaceutical herbal products > dietary supplements > traditional herbal remedies. The daily intake dose (DID) of pharmaceutical herbal products was within the established limits for the five metals. Dietary supplements and traditional herbal remedies exceeded the DID limits for Pb. The hazard quotients estimation and non-carcinogenic cumulative hazard estimation index for Mn, As, and Cd indicated no human health risk. Our results suggest that products containing G. biloba for sale in Mexico are not a health risk.

scholarly journals Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1577
Author(s):  
Klaudia Kotecka-Majchrzak ◽  
Natalia Kasałka-Czarna ◽  
Agata Sumara ◽  
Emilia Fornal ◽  
Magdalena Montowska
Keyword(s):  
Limit Of Detection ◽  
Raw Materials ◽  
Meat Products ◽  
Food Products ◽  
Plant Raw Materials ◽  
Heat Stable ◽  
2S Albumins ◽  
Specific Proteins ◽  
Meat Species ◽  
Species Specific

Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.

Quantitative Detection of Species-Specific DNA in Feedstuffs and Fish Meals

2005 ◽  
Vol 68 (6) ◽  
pp. 1217-1221 ◽  
Author(s):  
PAVEL KRCMAR ◽  
EVA RENCOVA
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Dna Sequences ◽  
Real Time Pcr ◽  
Single Species ◽  
Absolute Quantification ◽  
Quantitative Detection ◽  
Vertebrate Species ◽  
Target Species ◽  
Species Specific

A sensitive and rapid method for the quantitative detection of bovine-, ovine-, swine-, and chicken-specific mitochondrial DNA sequences based on real-time PCR has been developed. The specificity of the primers and probes for real-time PCR has been tested using DNA samples of other vertebrate species that may also be present in rendered products. The quantitative detection was performed with dual-labeled probes (TaqMan) using absolute quantification with external standards of single species meat-and-bone meals. This method facilitates the detection of 0.01% of the target species–derived material in concentrate feed mixtures and fish meals.

scholarly journals DNA-based species detection capabilities using laser transmission spectroscopy

2013 ◽  
Vol 10 (78) ◽  
pp. 20120637 ◽  
Author(s):  
A. R. Mahon ◽  
M. A. Barnes ◽  
F. Li ◽  
S. P. Egan ◽  
C. E. Tanner ◽  
...  
Keyword(s):  
Invasive Species ◽  
Dna Detection ◽  
Field Application ◽  
Economic Damage ◽  
Oligonucleotide Probes ◽  
Target Species ◽  
Dreissena Bugensis ◽  
Species Detection ◽  
Transmission Spectroscopy ◽  
Species Specific

Early detection of invasive species is critical for effective biocontrol to mitigate potential ecological and economic damage. Laser transmission spectroscopy (LTS) is a powerful solution offering real-time, DNA-based species detection in the field. LTS can measure the size, shape and number of nanoparticles in a solution and was used here to detect size shifts resulting from hybridization of the polymerase chain reaction product to nanoparticles functionalized with species-specific oligonucleotide probes or with the species-specific oligonucleotide probes alone. We carried out a series of DNA detection experiments using the invasive freshwater quagga mussel ( Dreissena bugensis ) to evaluate the capability of the LTS platform for invasive species detection. Specifically, we tested LTS sensitivity to (i) DNA concentrations of a single target species, (ii) the presence of a target species within a mixed sample of other closely related species, (iii) species-specific functionalized nanoparticles versus species-specific oligonucleotide probes alone, and (iv) amplified DNA fragments versus unamplified genomic DNA. We demonstrate that LTS is a highly sensitive technique for rapid target species detection, with detection limits in the picomolar range, capable of successful identification in multispecies samples containing target and non-target species DNA. These results indicate that the LTS DNA detection platform will be useful for field application of target species. Additionally, we find that LTS detection is effective with species-specific oligonucleotide tags alone or when they are attached to polystyrene nanobeads and with both amplified and unamplified DNA, indicating that the technique may also have versatility for broader applications.

scholarly journals Features of vitality structure of planting of Ginkgo biloba L. plants in the Ukrainian North-East

2019 ◽  
pp. 66-71
Author(s):  
I.M. Kovalenko ◽  
H.O. Klymenko ◽  
R.A. Yaroschuk ◽  
Yahui Su
Keyword(s):  
Ginkgo Biloba ◽  
Raw Materials ◽  
Folk Medicine ◽  
Leaf Size ◽  
Climatic Conditions ◽  
North East ◽  
High Viability ◽  
Plant Experiment ◽  
Number Of Leaves ◽  
Growing Conditions

Today, there are about 12,000 plants in the world that have healing properties and are used in both traditional and folk medicine. One of these plants is Ginkgo biloba L. In recent years, interest in its cultivation has increased in Ukraine, and improving the technology of growing this plant in the Ukrainian North-East is a relevant problem. Studies of G. biloba plants growing in the experimental area of Sumy NAU were conducted. On the basis of morphometric analysis, a number of morphoparameters were measured (plant height, annual growth of shoots, number of leaves, leaf size and leaf area, phytomass of the shoots, phytomass of leaves and phytomass of the stem, diameter of the shoots). The vital analysis, as well as the variance, correlation and regression analyses were carried out. G. biloba seedlings up to 3–4 years of life with different growing technology have a height of 25–30 cm and form 13–17 leaves per plant. The totality of morphometric characteristics in all variants of the G. biloba plant experiment corresponded to an equilibrium population of equilibrium type, in which in close proportions individuals of all three vitality types are present: a, b, and c. But at the same time a higher proportion (40 %) of individuals of class "a" was in the variant with the cultivation of G. biloba in greenhouse. The smallest part of individuals of high viability (only 20 %) was formed by cultivation of G. biloba in open soil without protection of the agro-grid. The ecological-coenotic stability of G. biloba has been noted many times, a certain limitation of G. biloba cultivation may be that this plant is light loving and thermophilic, but the climatic conditions of the Ukrainian North-East are favorable for it. Complex studies have shown the prospects and feasibility of growing G. biloba in the conditions of t the Ukrainian North-East as medicinal raw materials. Despite the stressful growing conditions for G. biloba, this species is characterized by high stability and adaptability, which is confirmed by our comparative morphometric and vital analysis of plants. Given that the age of the seedlings is negligible, further studies to determine the adaptability of G. biloba plants to growing conditions are not only desirable but also necessary.

scholarly journals AN EXHAUSTIVE REVIEW ON HERBS USED IN THE MANAGEMENT OF DIABETES MELLITUS TYPE 2

2019 ◽  
pp. 5-11
Author(s):  
Salman Mau ◽  
Shakir Saleem ◽  
Vishwadeepak Kimothi ◽  
Vineet Joshi ◽  
Sanjay Singh
Keyword(s):  
Diabetes Mellitus ◽  
Raw Materials ◽  
Nigella Sativa ◽  
Low Cost ◽  
Herbal Medicines ◽  
Analytical Techniques ◽  
Herbal Drugs ◽  
Herbal Products ◽  
High Blood Glucose ◽  
Glucose Levels

Diabetes mellitus is one of the most common metabolic disorders associated with disturbed hormonal secretion. Diabetes is characterized by high blood glucose levels over a prolonged period of time. High sugar levels are due to abnormal metabolism of carbohydrates and lipids which is caused by absolute or relative insulin deficiency. Herbal medicines have been the highly esteemed source of medicine throughout the human history. Herbs are becoming more popular today because of their least side effects, holistic beliefs, easy availability and low cost. Individual herbal products and formulations are gaining popularity because of their quality manufacturing using modern analytical techniques and standardized raw materials. Herbal drugs are widely used for the treatment of diabetes worldwide in various dosage forms. India has a long list of native herbal drugs with scientifically proven blood sugar lowering properties. The seeds of Nigella sativa, Olea europaea, fruits of Aegle marmelos, Momordica charantia, Coccinia indica, Nigella sativa,Gymnema sylvestre leaves,whole plant of Pterocarpus marsupium, Syzygium cumini fruits, Swertia punicea, Urtica dioica, gum of Ferula assa-foetida and seeds of Trigonella foenum graecum were discussed along with their reported mechanisms of action. In this review paper an attempt has been made to give an overview of certain Indian plants which have shown their anti-diabetic activity in various pre-clinical studies.

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