Comprehensive analysis using DNA metabarcoding, SCAR marker based PCR assay, and HPLC unveils the adulteration in Brahmi herbal products.
Bacopa monnieri
Botanical adulteration
Centella asiatica
DNA metabarcoding
SCAR marker
Journal
Molecular biology reports
ISSN: 1573-4978
Titre abrégé: Mol Biol Rep
Pays: Netherlands
ID NLM: 0403234
Informations de publication
Date de publication:
Sep 2023
Sep 2023
Historique:
received:
17
03
2023
accepted:
28
06
2023
medline:
29
8
2023
pubmed:
3
8
2023
entrez:
2
8
2023
Statut:
ppublish
Résumé
Brahmi is one of the important nootropic botanicals, widely sold in the market, with the name "Brahmi'' being used to describe both Bacopa monnieri and Centella asiatica species. The Brahmi herbal products market is expanding; hence, economically motivated adulteration is highly prevalent. This study aimed to develop DNA-based methods, including SCAR marker-based PCR and metabarcoding, to authenticate Brahmi herbal products and compare these methods with HPLC. These methods have been validated using mock controls (in-house blended formulations). All targeted plant species in mock controls were detected successfully with all three methods, whereas, in market samples, only 22.2%, 55.6%, and 50.0% were found positive for Brahmi by PCR assay, DNA metabarcoding, and HPLC, respectively. Metabarcoding can detect the presence of non-labeled plants together with targeted species, which is an advantage over PCR assay or HPLC. SCAR marker-based PCR is a rapid and cost-effective method for detecting the presence of B. monnieri and C. asiatica. However, in this study, the success rate of PCR amplification was relatively low because the primers targeted either RAPD or ITS-based SCAR markers. HPLC assay, although an alternative, was unable to detect the presence of other botanicals, just like the SCAR marker-based PCR assay. On the other hand, metabarcoding can be utilized to identify the target plants, even in very small quantities, while also providing simulated identification of other botanicals. This study successfully addressed the need for quality control of Brahmi herbal products and provided the first-time report of DNA metabarcoding for such products.
Sections du résumé
BACKGROUND
BACKGROUND
Brahmi is one of the important nootropic botanicals, widely sold in the market, with the name "Brahmi'' being used to describe both Bacopa monnieri and Centella asiatica species. The Brahmi herbal products market is expanding; hence, economically motivated adulteration is highly prevalent.
METHODS AND RESULTS
RESULTS
This study aimed to develop DNA-based methods, including SCAR marker-based PCR and metabarcoding, to authenticate Brahmi herbal products and compare these methods with HPLC. These methods have been validated using mock controls (in-house blended formulations). All targeted plant species in mock controls were detected successfully with all three methods, whereas, in market samples, only 22.2%, 55.6%, and 50.0% were found positive for Brahmi by PCR assay, DNA metabarcoding, and HPLC, respectively. Metabarcoding can detect the presence of non-labeled plants together with targeted species, which is an advantage over PCR assay or HPLC.
CONCLUSION
CONCLUSIONS
SCAR marker-based PCR is a rapid and cost-effective method for detecting the presence of B. monnieri and C. asiatica. However, in this study, the success rate of PCR amplification was relatively low because the primers targeted either RAPD or ITS-based SCAR markers. HPLC assay, although an alternative, was unable to detect the presence of other botanicals, just like the SCAR marker-based PCR assay. On the other hand, metabarcoding can be utilized to identify the target plants, even in very small quantities, while also providing simulated identification of other botanicals. This study successfully addressed the need for quality control of Brahmi herbal products and provided the first-time report of DNA metabarcoding for such products.
Identifiants
pubmed: 37532919
doi: 10.1007/s11033-023-08653-5
pii: 10.1007/s11033-023-08653-5
doi:
Substances chimiques
DNA
9007-49-2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
7605-7618Subventions
Organisme : Gujarat State Biotechnology Mission (GSBTM)
ID : GSBTM/JDRD/584/2018/204
Informations de copyright
© 2023. The Author(s), under exclusive licence to Springer Nature B.V.
Références
Shinomol GK, Muralidhara MS, Bharath M (2011) Exploring the role of “Brahmi” (Bacopa monnieri and Centella asiatica) in brain function and therapy. Recent Pat Endocr Metab Immune Drug Discov 5:33–49. https://doi.org/10.2174/187221411794351833
doi: 10.2174/187221411794351833
pubmed: 22074576
Mathur D, Goyal K, Koul V, Anand A (2016) The molecular links of re-emerging therapy: a review of evidence of brahmi (Bacopa monniera). Front Pharmacol 7:44
doi: 10.3389/fphar.2016.00044
pubmed: 26973531
pmcid: 4778428
Bhat R, Kiran K, Arun AB, Karim AA (2010) Determination of mineral composition and heavy metal content of some nutraceutically valued plant products. Food Anal Methods 3:181–187. https://doi.org/10.1007/s12161-009-9107-y
doi: 10.1007/s12161-009-9107-y
Keshari P (2021) Controversy, adulteration and substitution: burning problems in ayurveda practices. Nat Med Plants. https://doi.org/10.5772/intechopen.98220
doi: 10.5772/intechopen.98220
Meena H, Pandey HK, Pandey P et al (2012) Evaluation of antioxidant activity of two important memory enhancing medicinal plants Baccopa monnieri and Centella asiatica. Indian J Pharmacol 44:114
doi: 10.4103/0253-7613.91880
pubmed: 22345883
pmcid: 3271514
Mahima K, Sunil Kumar KN, Rakhesh KV et al (2022) Advancements and future prospective of DNA barcodes in the herbal drug industry. Front Pharmacol 13:1–19. https://doi.org/10.3389/fphar.2022.947512
doi: 10.3389/fphar.2022.947512
Aguiar S, Borowski T (2013) Neuropharmacological review of the nootropic herb Bacopa monnieri. Rejuvenation Res 16:313–326. https://doi.org/10.1089/rej.2013.1431
doi: 10.1089/rej.2013.1431
pubmed: 23772955
pmcid: 3746283
Soumyanath A, Zhong Y-P, Yu X et al (2010) Centella asiatica accelerates nerve regeneration upon oral administration and contains multiple active fractions increasing neurite elongation in-vitro. J Pharm Pharmacol 57:1221–1229. https://doi.org/10.1211/jpp.57.9.0018
doi: 10.1211/jpp.57.9.0018
Singh RH, Narsimhamurthy K, Singh G (2008) Neuronutrient impact of Ayurvedic Rasayana therapy in brain aging. Biogerontology 9:369–374. https://doi.org/10.1007/s10522-008-9185-z
doi: 10.1007/s10522-008-9185-z
pubmed: 18931935
Niraj S, Varsha S (2019) Role of Medhya Rasayanas (nootropic drugs) in developmental disabilities of children. Ment Retard 25:30
Kulkarni R, Girish KJ, Kumar A (2012) Nootropic herbs (Medhya Rasayana) in Ayurveda: an update. Pharmacogn Rev 6:147–153. https://doi.org/10.4103/0973-7847.99949
doi: 10.4103/0973-7847.99949
pubmed: 23055641
pmcid: 3459457
Goraya GS, Ved DK Medicinal plants in India: an assessment of their demand and supply. Natl Med Plants Board, Minist AYUSH (2017) Gov India, New Delhi Indian Counc For Res Educ Dehradun 100–105
Abdul Manap AS, Vijayabalan S, Madhavan P et al (2019) Bacopa monnieri, a neuroprotective lead in Alzheimer disease: a review on its properties, mechanisms of action, and preclinical and clinical studies. Drug Target Insights 13:1177392819866412. https://doi.org/10.1177/1177392819866412
doi: 10.1177/1177392819866412
pubmed: 31391778
pmcid: 6669844
Brahmi Market Forecast (2021–2026) Industry ARC. https://www.industryarc.com/ Research/Brahmi- Market-Research-507354
Yadav A, Ahmad J, Chaudhary AA, Ahmad A (2012) Development of sequence characterized amplified region (SCAR) marker for the authentication of Bacopa monnieri (L.) Wettst. Eur J Med Plants 2:186–198
doi: 10.9734/EJMP/2012/1192
Balaji R, Parani M (2022) DNA barcoding of the market samples of single-drug herbal powders reveals adulteration with taxonomically unrelated plant species. Diversity. https://doi.org/10.3390/d14060495
doi: 10.3390/d14060495
Alqahtani A, Cho J-L, Wong KH et al (2017) Differentiation of three Centella species in Australia as inferred from morphological characteristics, ISSR molecular fingerprinting and phytochemical composition. Front Plant Sci 8:1980
doi: 10.3389/fpls.2017.01980
pubmed: 29209345
pmcid: 5702339
Pant P, Pandey S, Dall’Acqua S (2021) The influence of environmental conditions on secondary metabolites in medicinal plants: a literature review. Chem Biodivers 18:e2100345
doi: 10.1002/cbdv.202100345
pubmed: 34533273
Raclariu AC, Heinrich M, Ichim MC, de Boer H (2018) Benefits and limitations of DNA barcoding and metabarcoding in herbal product authentication. Phytochem Anal 29:123–128. https://doi.org/10.1002/pca.2732
doi: 10.1002/pca.2732
pubmed: 28906059
Newmaster SG, Grguric M, Shanmughanandhan D et al (2013) DNA barcoding detects contamination and substitution in north american herbal products. BMC Med 11:222. https://doi.org/10.1186/1741-7015-11-222
doi: 10.1186/1741-7015-11-222
pubmed: 24120035
pmcid: 3851815
Wu HY, Shaw PC (2022) Strategies for molecular authentication of herbal products: from experimental design to data analysis. Chin Med (United Kingdom) 17:1–15. https://doi.org/10.1186/s13020-022-00590-y
doi: 10.1186/s13020-022-00590-y
Sharma S, Shrivastava N (2016) DNA-based simultaneous identification of three Terminalia species targeting adulteration. Pharmacogn Mag 12:S379–S383. https://doi.org/10.4103/0973-1296.185776
doi: 10.4103/0973-1296.185776
pubmed: 27563228
pmcid: 4971960
Zhang T, Xu F, Ruhsam M et al (2022) A nucleotide signature for the identification of Pinelliae Rhizoma (Banxia) and its products. Mol Biol Rep 49:7753–7763. https://doi.org/10.1007/s11033-022-07600-0
doi: 10.1007/s11033-022-07600-0
pubmed: 35670929
pmcid: 9171473
Travadi T, Sharma S, Pandit R et al (2022) A duplex PCR assay for authentication of Ocimum basilicum L. and Ocimum tenuiflorum L in Tulsi churna. Food Control 137:108790. https://doi.org/10.1016/j.foodcont.2021.108790
doi: 10.1016/j.foodcont.2021.108790
Travadi T, Shah AP, Pandit R et al (2022) Detection of Carica papaya adulteration in Piper nigrum using chloroplast DNA marker-based PCR assays. Food Anal Methods. https://doi.org/10.1007/s12161-022-02395-z
doi: 10.1007/s12161-022-02395-z
Vassou SL, Nithaniyal S, Raju B, Parani M (2016) Creation of reference DNA barcode library and authentication of medicinal plant raw drugs used in ayurvedic medicine. BMC Complement Altern Med 16. https://doi.org/10.1186/s12906-016-1086-0
Intharuksa A, Sasaki Y, Ando H et al (2019) The combination of ITS2 and psba-trnh region is powerful DNA barcode markers for authentication of medicinal Terminalia plants from Thailand. J Nat Med. https://doi.org/10.1007/s11418-019-01365-w
doi: 10.1007/s11418-019-01365-w
pubmed: 31587135
Tanaka S, Ito M (2020) DNA barcoding for identification of agarwood source species using trnl-trnf and matK DNA sequences. J Nat Med 74:42–50. https://doi.org/10.1007/s11418-019-01338-z
doi: 10.1007/s11418-019-01338-z
pubmed: 31250369
Thakur VV, Tripathi N, Tiwari S (2021) DNA barcoding of some medicinally important plant species of Lamiaceae family in India. Mol Biol Rep 48:3097–3106. https://doi.org/10.1007/s11033-021-06356-3
doi: 10.1007/s11033-021-06356-3
pubmed: 33913093
Pang X, Shi L, Song J et al (2013) Use of the potential DNA barcode ITS2 to identify herbal materials. J Nat Med 67:571–575. https://doi.org/10.1007/s11418-012-0715-2
doi: 10.1007/s11418-012-0715-2
pubmed: 23179313
Seethapathy GS, Raclariu-Manolica A-C, Anmarkrud JA et al (2019) DNA metabarcoding authentication of ayurvedic herbal products on the european market raises concerns of quality and fidelity. Front Plant Sci 10:68
doi: 10.3389/fpls.2019.00068
pubmed: 30804961
pmcid: 6370972
Raclariu AC, Ţebrencu CE, Ichim MC et al (2018) What’s in the box? Authentication of Echinacea herbal products using DNA metabarcoding and HPTLC. Phytomedicine 44:32–38. https://doi.org/10.1016/j.phymed.2018.03.058
doi: 10.1016/j.phymed.2018.03.058
pubmed: 29895490
Arulandhu AJ, Staats M, Hagelaar R et al (2017) Development and validation of a multi-locus DNA metabarcoding method to identify endangered species in complex samples. Gigascience. https://doi.org/10.1093/gigascience/gix080
doi: 10.1093/gigascience/gix080
pubmed: 29020743
pmcid: 5632295
Frigerio J, Agostinetto G, Mezzasalma V et al (2021) DNA-based herbal teas’ authentication: an ITS2 and psba‐trnh multi‐marker DNA metabarcoding approach. Plants 10:1–14. https://doi.org/10.3390/plants10102120
doi: 10.3390/plants10102120
Travadi T, Shah AP, Pandit R et al (2023) A combined approach of DNA metabarcoding collectively enhances the detection efficiency of medicinal plants in single and polyherbal formulations. Front Plant Sci 14:1169984
doi: 10.3389/fpls.2023.1169984
pubmed: 37255553
pmcid: 10225634
Raclariu AC, Mocan A, Popa MO et al (2017) Veronica officinalis product authentication using DNA metabarcoding and HPLC-MS reveals widespread adulteration with Veronica chamaedrys. Front Pharmacol 8:378
doi: 10.3389/fphar.2017.00378
pubmed: 28674497
Sánchez M, González-Burgos E, Divakar PK, Gómez-Serranillos MP (2020) DNA-based authentication and metabolomics analysis of medicinal plants samples by DNA barcoding and ultra-high-performance liquid chromatography/triple quadrupole mass spectrometry (UHPLC-MS). Plants 9:1601
doi: 10.3390/plants9111601
pubmed: 33218119
pmcid: 7698941
The Indian Pharmacopoeia Commission (2007) Indian pharmacopoeia. Vol III
Maloukh L, Kumarappan A, Jarrar M et al (2017) Discriminatory power of rbcL barcode locus for authentication of some of United Arab Emirates (UAE) native plants. 3 Biotech 7:1–7
doi: 10.1007/s13205-017-0746-1
Travadi T, Sharma S, Pandit R et al (2022) A duplex PCR assay for authentication of Ocimum basilicum L. and Ocimum tenuiflorum L. in Tulsi churna. Food Control. https://doi.org/10.1016/j.foodcont.2021.108790
doi: 10.1016/j.foodcont.2021.108790
Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386. https://doi.org/10.1385/1-59259-192-2:365
doi: 10.1385/1-59259-192-2:365
pubmed: 10547847
Kibbe WA (2007) OligoCalc: an online oligonucleotide properties calculator. Nucleic Acids Res 35:43–46. https://doi.org/10.1093/nar/gkm234
doi: 10.1093/nar/gkm234
Dormontt EE, Van Dijk K-J, Bell KL et al (2018) Advancing DNA barcoding and metabarcoding applications for plants requires systematic analysis of herbarium collections—an australian perspective. Front Ecol Evol 6:134
doi: 10.3389/fevo.2018.00134
Schmieder R, Edwards R (2011) Quality control and preprocessing of metagenomic datasets. Bioinformatics 27:863–864. https://doi.org/10.1093/bioinformatics/btr026
doi: 10.1093/bioinformatics/btr026
pubmed: 21278185
pmcid: 3051327
Huang Y, Niu B, Gao Y et al (2010) CD-HIT suite: a web server for clustering and comparing biological sequences. Bioinformatics 26:680–682. https://doi.org/10.1093/bioinformatics/btq003
doi: 10.1093/bioinformatics/btq003
pubmed: 20053844
pmcid: 2828112
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
doi: 10.1016/S0022-2836(05)80360-2
pubmed: 2231712
Deepak M, Sangli GK, Arun PC, Amit A (2005) Quantitative determination of the major saponin mixture bacoside A in Bacopa monnieri by HPLC. Phytochem Anal 16:24–29. https://doi.org/10.1002/pca.805
doi: 10.1002/pca.805
pubmed: 15688952
Urumarudappa SKJ, Tungphatthong C, Prombutara P, Sukrong S (2020) DNA metabarcoding to unravel plant species composition in selected herbal medicines on the national list of essential medicines (NLEM) of Thailand. Sci Rep 10:1–11. https://doi.org/10.1038/s41598-020-75305-0
doi: 10.1038/s41598-020-75305-0
Anthoons B, Karamichali I, Schrøder-Nielsen A et al (2021) Metabarcoding reveals low fidelity and presence of toxic species in short chain-of-commercialization of herbal products. J Food Compos Anal 97:103767
doi: 10.1016/j.jfca.2020.103767
Ichim MC (2019) The DNA-based authentication of commercial herbal products reveals their globally widespread adulteration. Front Pharmacol 10:1–9. https://doi.org/10.3389/fphar.2019.01227
doi: 10.3389/fphar.2019.01227
Raclariu AC, Paltinean R, Vlase L et al (2017) Comparative authentication of Hypericum perforatum herbal products using DNA metabarcoding, TLC and HPLC-MS. Sci Rep 7:8–10. https://doi.org/10.1038/s41598-017-01389-w
doi: 10.1038/s41598-017-01389-w
Bansal A, Chhabra V, Rawal RK, Sharma S (2014) Chemometrics: a new scenario in herbal drug standardization. J Pharm Anal 4:223–233
doi: 10.1016/j.jpha.2013.12.001
pubmed: 29403886
pmcid: 5761221
Taberlet P, Coissac E, Pompanon F et al (2007) Power and limitations of the chloroplast trn L (UAA) intron for plant DNA barcoding. Nucleic Acids Res 35:e14–e14
doi: 10.1093/nar/gkl938
pubmed: 17169982
Little DP (2014) A DNA mini-barcode for land plants. Mol Ecol Resour 14:437–446
doi: 10.1111/1755-0998.12194
pubmed: 24286499
Zhu S, Liu Q, Qiu S et al (2022) DNA barcoding: an efficient technology to authenticate plant species of traditional chinese medicine and recent advances. Chin Med 17:1–17
doi: 10.1186/s13020-022-00655-y
Hebert PDN, Cywinska A, Ball SL, DeWaard JR (2003) Biological identifications through DNA barcodes. Proc R Soc B Biol Sci 270:313–321. https://doi.org/10.1098/rspb.2002.2218
doi: 10.1098/rspb.2002.2218
de Boer HJ, Ichim MC, Newmaster SG (2015) DNA barcoding and pharmacovigilance of herbal medicines. Drug Saf 38:611–620. https://doi.org/10.1007/s40264-015-0306-8
doi: 10.1007/s40264-015-0306-8
pubmed: 26076652
Seethapathy GS, Ganesh D, Santhosh Kumar JU et al (2015) Assessing product adulteration in natural health products for laxative yielding plants, Cassia, Senna, and Chamaecrista, in Southern India using DNA barcoding. Int J Legal Med 129:693–700. https://doi.org/10.1007/s00414-014-1120-z
doi: 10.1007/s00414-014-1120-z
pubmed: 25425095
Sharma S, Shrivastava N (2016) Internal transcribed spacer guided multiplex PCR for species identification of Convolvulus prostratus and Evolvulus alsinoides. Acta Pharm Sin B 6:253–258. https://doi.org/10.1016/j.apsb.2016.02.003
doi: 10.1016/j.apsb.2016.02.003
pubmed: 27175337
pmcid: 4856953
Kumar A, Mishra P, Baskaran K et al (2016) Higher efficiency of ISSR markers over plastid psba-trnh region in resolving taxonomical status of genus Ocimum L. Ecol Evol 6:7671–7682
doi: 10.1002/ece3.2483
pubmed: 30128120
pmcid: 6093152
Xin T, Xu Z, Jia J et al (2018) Biomonitoring for traditional herbal medicinal products using DNA metabarcoding and single molecule, real-time sequencing. Acta Pharm Sin B 8:488–497. https://doi.org/10.1016/j.apsb.2017.10.001
doi: 10.1016/j.apsb.2017.10.001
pubmed: 29881688
Negi RK, Nautiyal P, Bhatia R, Verma R (2021) rbcL, a potential candidate DNA barcode loci for aconites: conservation of himalayan aconites. Mol Biol Rep 48:6769–6777. https://doi.org/10.1007/s11033-021-06675-5
doi: 10.1007/s11033-021-06675-5
pubmed: 34476739
Kiran KR, Swathy PS, Paul B et al (2021) Untargeted metabolomics and DNA barcoding for discrimination of Phyllanthus species. J Ethnopharmacol 273:113928. https://doi.org/10.1016/j.jep.2021.113928
doi: 10.1016/j.jep.2021.113928
pubmed: 33631274
Zhang X, Zhou T, Yu W et al (2018) Development and evaluation of a PCR-based assay kit for authentication of Zaocys dhumnades in traditional chinese medicine. Mitochondrial DNA Part A DNA Mapp Seq Anal 29:102–106. https://doi.org/10.1080/24701394.2016.1248429
doi: 10.1080/24701394.2016.1248429
Noh P, Kim WJ, Yang S et al (2021) PCR-based rapid diagnostic tools for the authentication of medicinal mistletoe species. Phytomedicine 91:153667. https://doi.org/10.1016/j.phymed.2021.153667
doi: 10.1016/j.phymed.2021.153667
pubmed: 34332281
Coghlan ML, Haile J, Houston J et al (2012) Deep sequencing of plant and animal DNA contained within traditional Chinese medicines reveals legality issues and health safety concerns. PLoS Genet. https://doi.org/10.1371/journal.pgen.1002657
doi: 10.1371/journal.pgen.1002657
pubmed: 22511890
pmcid: 3325194
Yao Q, Zhu X, Han M et al (2022) Decoding herbal materials of TCM preparations with the multi-barcode sequencing approach. Sci Rep 12:1–18. https://doi.org/10.1038/s41598-022-09979-z
doi: 10.1038/s41598-022-09979-z
Cheng X, Su X, Chen X et al (2014) Biological ingredient analysis of traditional chinese medicine preparation based on high-throughput sequencing: the story for Liuwei Dihuang Wan. Sci Rep 4:1–12
doi: 10.1038/srep05147
Ivanova NV, Kuzmina ML, Braukmann TWA et al (2016) Authentication of herbal supplements using next-generation sequencing. PLoS ONE 11:1–24. https://doi.org/10.1371/journal.pone.0156426
doi: 10.1371/journal.pone.0156426
Loman NJ, Misra RV, Dallman TJ et al (2012) Performance comparison of benchtop high-throughput sequencing platforms. Nat Biotechnol 30:434–439. https://doi.org/10.1038/nbt.2198
doi: 10.1038/nbt.2198
pubmed: 22522955
Salipante SJ, Kawashima T, Rosenthal C et al (2014) Performance comparison of Illumina and Ion Torrent next-generation sequencing platforms for 16S rRNA-based bacterial community profiling. Appl Environ Microbiol 80:7583–7591. https://doi.org/10.1128/AEM.02206-14
doi: 10.1128/AEM.02206-14
pubmed: 25261520
pmcid: 4249215
Pawluczyk M, Weiss J, Links MG et al (2015) Quantitative evaluation of bias in PCR amplification and next-generation sequencing derived from metabarcoding samples. Anal Bioanal Chem 407:1841–1848. https://doi.org/10.1007/s00216-014-8435-y
doi: 10.1007/s00216-014-8435-y
pubmed: 25577362
Staats M, Arulandhu AJ, Gravendeel B et al (2016) Advances in DNA metabarcoding for food and wildlife forensic species identification. Anal Bioanal Chem 408:4615–4630. https://doi.org/10.1007/s00216-016-9595-8
doi: 10.1007/s00216-016-9595-8
pubmed: 27178552
pmcid: 4909793
Hinchliff CE, Smith SA (2014) Some limitations of public sequence data for phylogenetic inference (in plants). PLoS ONE. https://doi.org/10.1371/journal.pone.0098986
doi: 10.1371/journal.pone.0098986
pubmed: 24999823
pmcid: 4085032
Liu C, Guo D, Liu L (2018) Quality transitivity and traceability system of herbal medicine products based on quality markers. Phytomedicine 44:247–257. https://doi.org/10.1016/j.phymed.2018.03.006
doi: 10.1016/j.phymed.2018.03.006
pubmed: 29631807