Alternative approach to species identification of Actaea racemosa L. (syn. Cimicifuga racemosa (L.) Nutt., black cohosh) herbal starting material: UV spectroscopy coupled with LDA

The North American Actaea racemosa L. (syn. Cimicifuga racemosa (L.) Nutt., commonly known as black cohosh), considered to be a more “natural” alternative to conventional therapies, is used to relieve menopausal symptoms. The high demand for plant material has led to problems with substitution/adulteration of raw material of wholesale origin. The authenticity of the starting material is crucial for the herbal product’s efficacy and safety, and tests on identity and substitution are integral parts of cGMP guidelines. Consequently, there is a need for economical and easy-applicable test procedures. The aim of this study is to reveal the capability of the well-established UV spectroscopy coupled with a multivariate classification procedure to serve as a tool for the identification of A. racemosa. We built a classification model applying linear discriminant analysis (LDA) to distinguish between A. racemosa and its common substitutes. The model showed a high level of accuracy predicting 100% of the samples correctly. Our results indicate that UV spectroscopy shows potential for the development of possible additional authentication methods for this herbal starting material.     

Photopyroelectric Detection of Vegetable Oils' Adulteration

Highly accurate photopyroelectric (PPE) investigation, in the thermal-wave-resonator-cavity configuration, was combined with gas chromatography (GC), in order to detect adulteration of flax oil by mixing with sunflower oil. It was found that the value of the thermal diffusivity for the investigated mixtures ranges from 8.07 × 10⁸m²/s (pure sunflower oil) to 10.03 × 10⁸m²/s (pure flax oil) and is directly correlated with the total amount of the polyunsaturated fatty acids from oils composition. The correlation between the PPE signal and the composition of unadulterated and adulterated vegetable oils confirms the data obtained previously on fresh and spoiled vegetable oils and suggests that the thermal diffusivity may be a suitable parameter to detect oils' quality and their early spoilage and adulteration.     

Physicochemical and adulteration study of fresh milk collected from different locations in Pakistan

The present research was carried out to assess raw milk's quality as collected from the commercial markets of the Mardan district (Khyber Pakhtunkhwa). The locality from which milk samples were collected included; Bijligar (BG), Manga (M), Chamthara (CM), Main Bazar Mardan (MB), Mahidherai (MD), and Sharif Abad (SB), located in district Mardan, Khyber Pakhtunkhwa (KP). A total of 36 milk samples were collected at the rate of 6 samples per location. The outcome of the data exhibited that the percentage of protein content was highest (3.34%) in MB and SA (3.30%), while lower percentages were recorded in sample M (3.03%) and CM 93.06%). Maximum pH were shown in M and BG to be 7.55 and 7.33, respectively. For fats content, the highest percentage of fats was witnessed in MB as 4.04%, and minimum fats content was noted in Sample M as 3.57%. Water content was highest in Sample SA and BG at 15.85% and 15.64%, respectively. Qualitative analysis of adulterants like detergents, Formalin, starch, and Hydrogen peroxide was also carried out for all the collected samples. Adulteration results were positive for all the milk samples, with the highest being in samples MB (30%), while all the remaining samples had adulteration at 20% each. Both MB and CM samples were adulterated with urea, while the remaining 4 were adulterated with neutralizers. Thus, it may be summarized from the whole analysis that the milk available in commercial markets of district Mardan was adulterated with different adulterants and is not recommended for consumption.     

Nostoc flagelliforme and faked items retailed in Hong Kong

A survey was made to check the authenticity of both dried and cookedNostoc flagelliforme retailed as Fat Choy in Hong Kong,using microscopic and histochemical methods. Results indicated that faked itemswere found in 70% of the 30 samples of dried Fat Choy retailed in seafoodstoresand herb shops and in all 5 samples of cooked Fat Choy obtained from Chineserestaurants. The faked items were non-cellular and packed with starch grains ormasses and black pigments. Staining the faked items with iodine solution turnthem into dark blue or black, whereas the genuine samples remained dullgreenish. This widespread adulteration probably reflects the limited supply ofthis alga which is banned in China from further collection and trading.     

Specific,sensitive and rapid Curcuma longa turmeric powder authentication in commercial food using loop-mediated isothermal nucleic acid amplification

Turmeric (Curcuma longa) is a rhizomatous plant of the ginger family Zingiberaceae that is usually dried and ground into powder for use as a seasoning. Because turmeric has become increasingly popular in the functional food market, adulteration of C. longa by other turmeric species is becoming an increasingly significant problem. In this study, loop-mediated isothermal amplification (LAMP) was developed for the detection of C. longa DNA for turmeric authentication. ITS2-26S rDNA was used for the LAMP primer designation. The results demonstrated that the specific primers exhibited high specificity, authenticated C. longa DNA within 30 min at 65 °C isothermally and had no cross-reaction with other adulterants. LAMP was sensitive to 0.1 ng of turmeric C. longa DNA, and only 0.01% of C. longa turmeric powder in the sample was required for DNA amplification. The sensitivity of LAMP was 10-fold higher than that of PCR (0.1%) from a previous report. Moreover, all the collected commercial turmeric products were positively detected by LAMP and RtF-LAMP (real-time fluorescence LAMP). The developed LAMP assay not only had higher specificity and rapidity than that of other methods but could also be applied to authenticate turmeric to prevent adulteration in food products.     

High performance thin layer chromatography as a method to authenticate Hoodia gordonii raw material and products

Hoodia gordonii which contains the perceived active molecule, P57, is a plant used in many weight loss products that are highly susceptible to adulteration due to increased public demand and limited availability. Rapid and simple methods for authentication and confirmation of the presence of P57 are desirable for the quality control of H. gordonii raw material and products. High performance thin layer chromatography (HPTLC) analysis of several H. gordonii raw material samples collected from different locations as well as weight loss products was carried out on silica gel plates and developed in a mobile phase of toluene:chloroform:ethanol (40:40:12.5 v/v/v). Liebermann–Burchard (LB) reagent was used as derivatising agent since it is specific for glycosides and triterpenes (such as P57) and the plates were viewed under UV light at 365 nm. This method produced good separation of the compounds in complex mixtures with well-defined bands including that of the P57 band (R_f 0.42), which was confirmed by liquid chromatography coupled to mass spectrometry (LC–MS) after preparative thin layer chromatography (TLC). All the HPTLC results obtained for the H. gordonii raw materials and products were confirmed with quantitative LC–MS analyses, which confirmed the qualitative reliability of the HPTLC method. The HPTLC method was used successfully to develop a chemical fingerprint for authentication and reliable confirmation of the presence of P57 in H. gordonii raw material and products.     

Authentication of the medicinal plant Sennaangustifolia by RAPD profiling

In this study “RAPD” molecular marker was employed for the identification of Sennaangustifolia, Sennaacutifolia, Sennatora and Sennasophera. Total 32 decamer primers were screened in amplification with genomic DNA extracted from all species, of which 6 primers yielded species-specific reproducible bands. Out of 42 loci detected, the polymorphic, monomorphic and unique loci were 24, 2 and 16, respectively. Based on dendrogram and similarity matrix, 4 species were differentiated from each other and showed more divergence. Thus, this technique may prove and to contribute the identification of these species of Senna having similar morphology sold in the local markets.     

Adulteration of Ginkgo biloba products and a simple method to improve its detection

Extracts of ginkgo (Ginkgo biloba) leaf are widely available worldwide in herbal medicinal products, dietary supplements, botanicals and complementary medicines, and several pharmacopoeias contain monographs for ginkgo leaf, leaf extract and finished products. Being a high-value botanical commodity, ginkgo extracts may be the subject of economically motivated adulteration. We analysed eight ginkgo leaf retail products purchased in Australia and Denmark and found compelling evidence of adulteration with flavonol aglycones in three of these. The same three products also contained genistein, an isoflavone that does not occur in ginkgo leaf.Although the United States Pharmacopeia – National Formulary (USP-NF) and the British and European Pharmacopoeias stipulate a required range for flavonol glycosides in ginkgo extract, the prescribed assays quantify flavonol aglycones. This means that these pharmacopoeial methods are not capable of detecting adulteration of ginkgo extract with free flavonol aglycones.We propose a simple modification of the USP-NF method that addresses this problem: by assaying for flavonol aglycones pre and post hydrolysis the content of flavonol glycosides can be accurately estimated via a simple calculation. We also recommend a maximum limit be set for free flavonol aglycones in ginkgo extract.     

Adulteration identification of some fungal polysaccharides with SEM, XRD, IR and optical rotation: A primary approach

Edible mushrooms have been consumed in Asian countries for healthy functions historically with polysaccharides, which have been manufactured as ending products, as one of the major bioactive components. Driven by profits, some inglorious manufacturers conducted adulteration using cheap carbohydrates. The extracted polysaccharides from Agaricus blazei Murill, Ganoderma lucidum and Lentinus edodes were chosen as products and maltodextrin, carboxymethyl cellulose (CMC), water soluble corn starch and guar gum as adulterants for adulteration investigation. Scanning electron microscope (SEM) observation implied regular granular shapes of fungal polysaccharides different significantly from the adulterants. Infrared spectra revealed α- and β-anomeric types of polysaccharides and carbohydrates. X-ray diffractometry (XRD) suggested different diffraction patterns of polysaccharides from adulterant carbohydrates, the former were amorphous while the later somewhat crystalline of different crystallinity. Chiral properties of polysaccharides and carbohydrates showed big differences in specific rotation with good correlation between the content of “pure” fungal polysaccharides and adulterants.