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. 2017 Jul 6;7:4774. doi: 10.1038/s41598-017-05083-9

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Workflow for quantitative detection of saffron in crude drugs or in Tibetan medicine “Ershiwuwei Shanhu Wan”. Step 1. Dry the saffron samples or coarse powder of “Ershiwuwei Shanhu Wan” at 60 °C for 2 hours to remove excessive water. Step 2. Grind the dried samples into powder. Step 3. Pass the powder through a 100 mesh sieve (saffron powder used for Ershiwuwei Shanhu Wan recovery test was passed through a 200 mesh sieve as it was used in the industrial production) to get homogenized samples. Step 4. Extract the DNA by direct plant DNA extraction solution. Step 5. Determine the saffron in crude drugs or Tibetan medicine “Ershiwuwei Shanhu Wan” by quantifying species-specific DNA through real-time PCR.