Identification of adulteration in the market samples of saffron using morphology, HPLC, HPTLC, and DNA barcoding methods.

Saffron, the stigma of Crocus sativus L., is the most expensive spice used for culinary, medicinal, dye, and cosmetics purposes. It is highly adulterated because of its limited production and high commercial value. In this study, 104 saffron market samples collected from 16 countries were tested using morphology, high-performance liquid chromatography (HPLC), high-performance thin-layer chromatography (HPTLC), and deoxyribonucleic acid (DNA) barcoding. Overall, 45 samples (43%) were adulterated. DNA barcoding identified the highest number of adulterated saffron (44 samples), followed by HPTLC (39 samples), HPLC (38 samples), and morphology (32 samples). Only DNA barcoding identified the adulterated samples containing saffron and other plants' parts as bulking agents. In addition, DNA barcoding identified 20 adulterant plant species, which will help develop quality control methods and market surveillance. Some of the adulterant plants are unsafe for human consumption. The HPLC method helped identify the saffron samples adulterated with synthetic safranal. HPLC and HPTLC methods will help identify the samples adulterated with other parts of the saffron plant (auto-adulteration).

Identification of synthetic dyes magenta III (new fuchsin) and rhodamine B as common adulterants in commercial saffron.

Saffron is a highly adulterated spice due to its limited production and high costs. Non-saffron plant material is coloured with synthetic dyes to produce counterfeit saffron. Continuous monitoring of the synthetic dyes used in counterfeit saffron is essential because some dyes are not safe for human consumption. In the present study, 104 commercial saffron samples from 16 countries were screened, and 20 samples were found to contain dyes. Thin-Layer Chromatography (TLC) analysis showed adulteration with magenta- and pink- coloured dyes (18 and 2 samples, respectively), which did not correspond to any of the dyes reported earlier. Mass spectrometry analysis identified the magenta-coloured dye as magenta III or new fuchsin and the pink-coloured dye as rhodamine B. Magenta III is regarded as possibly carcinogenic, and rhodamine B was reported to cause local sarcomas. Both of these synthetic dyes are reported as adulterants in saffron for the first time.

Authentication of the market samples of Ashwagandha by DNA barcoding reveals that powders are significantly more adulterated than roots.

ETHNOPHARMACOLOGICAL RELEVANCE: Ashwagandha, also known as Indian Ginseng, is a highly traded medicinal plant, which is used in Ayurveda, Siddha and Unani systems of medicine to improve cognitive function, decrease inflammation, and to counter the ill-effects of aging. Withanolide A and Withaferin A from Ashwagandha were shown to improve immunity and have anti-cancer property, respectively. AIM OF THE STUDY: Here, we aimed to create reference DNA barcodes for W. somnifera and to authenticate root and powder samples of Ashwagandha collected from markets. MATERIALS AND METHODS: Three plant specimen of W. somnifera were collected, and reference DNA barcodes were generated using rbcL, matK, trnH-psbA, and ITS2 DNA barcode markers. Market samples in the form of root (n = 33) and powder (n = 70) were collected and authenticated using ITS2 and trnH-psbA DNA barcodes. RESULTS: Genomic DNA was successfully isolated from all plant specimens and market samples. DNA barcoding showed that 77% of samples were authentic. About 22% of non-authentic samples were powder samples and only 1% were root samples. Among the non-authentic samples, 18% were completely substituted with single species (Mucuna pruriens (L.) DC., Trigonella foenum-graceum L., or Senna auriculata (L.) Roxb.) and 82% were mixed samples containing more than one species. About 63% of the mixed samples contained Ashwagandha as the major ingredient. Furthermore, we identified that six taxonomically divergent plant species from four families were present as adulterants in the mixed samples. CONCLUSION: DNA barcoding revealed that botanical adulteration in the market samples of Ashwagandha is significant. Powder samples are more prone to adulteration than root samples. The adulterated samples contained plant material that is not related to Ashwagandha, which warrants strict quantity control and market surveillance to derive the true medicinal benefits of this medicinal plant.