BEWARE OF FAKE DRUGS:

An example from Antidesma bunius (L.) Spreng. (Bignay) Products in the

Baay, Lord Mikhael S., Catamio, MarcPhilippine
Exequiel L., Estrella,Market
Paul Janvert R., Rodriguez, Gabriel Dominic A.,
Binag, Sam Dominic A., Alfeche, Niña Kathryn G., and Alejandro, Grecebio Jonathan D.
Department of Biological Sciences, University of Santo Tomas, Espana Blvd.., 1015 Manila, Philippines

INTRODUCTION RESULTS AND DISCUSSION

• Commercially available Antidesma bunius (L.) Spreng., (Fig. 1) locally known as B. Herbal Product Authentication
“bignay”, is widely used in the Philippines for its medicinal benefits.
• Its fruits (Fig. 1B) contain flavonoids which are powerful antioxidants with anti- The gathered HMPs must satisfy two criteria for authentication: (1) BLASTn,
inflammatory and immune system benefits (Butkhup & Samappito, 2008). and (2) ML TREE RECONSTRUCTION. The Antidesma bunius – derived HMP
• Products of bignay are now sold throughout the public market but without the must have a BLASTn ID of A. bunius with an e value < 0.01 for all of the tested
standard quality control (Tshitenge et al., 2016). loci to be considered authentic (Table 2). It must also nest within the established A.
• The World Health Organization (WHO) reported that the presence of substitutes bunius clade (with BS >70%) in the SRM herbal barcode library (Figure 3).
and contaminants in the adulterated Herbal Medicinal Products (HMPs) might
cause possible harmful effects (WHO, 2017). Table 2. BLASTn results of barcode sequences generated from the herbal products.
• WHO has raised the awareness of quality control in these HMPs by the application
of modern techniques, such as the DNA barcoding.
OBJECTIVES:
• Establish the distinctness of A. bunius from its close allies following DNA barcoding
multi-locus/tiered approach using matK, rbcL, psbA-trnH markers.
• Authenticate at least 10 bignay derived HMPs following the BLASTn criterion and
Maximum-likelihood (ML) tree construction criterion.
S I G N I F I C A N C E:
• The establishment of a standardized authentication protocol will ensure
consumer health by preventing the presence of adulterants/contaminants and
fraudulent products in the Philippine market.

A B C
Figure 1. (A) Buds, (B) fruits, and (C) leaves of A. bunius
MATERIALS AND METHODS
Herbal Product
Establishment of Standards
Authentication

COLLECTION OF A. bunius COLLECTION OF A. bunius Figure 3. Maximum-Likelihood tree for Antidesma bunius - derived HMPs using the multi-
SAMPLES SAMPLES locus/tiered approach. Highlighted is the authenticated A. bunius products while samples
9 wild bignay samples 11 bignay derived HMPs coded in RED are considered adulterated/substituted.

matK, rbcL & trnH-psbA matK, rbcL & trnH-psbA Of the 11 herbal medicinal products tested bought from prominent markets
Barcodes and Generation Barcodes and Generation
in the country, ONLY THREE (3) were considered AUTHENTIC A. bignay –
derived products.
Sequence Characteristics BLASTn
Computation (e-value<0.01)
CONCLUSION
ML Tree Criterion DNA barcoding protocols were able to establish the Standard Reference
Establishment of Standard
Reference Material (SRM)
Considered Authentic if Material (SRM) Herbal barcode library for Antidesma bunius and its related
Monophyletic w/ A. bunius
species. the SRM clearly delineated the identity of the wild samples and was able
RESULTS AND DISCUSSION to authenticate the bignay – derived products that are sold in the
Philippine market. The testing revealed that majority of the products contained
A. Standard Reference Material herbal barcode library for Antidesma spp. adulterants which could pose health threats to the consumers.
The SRM (Figure 2) establishes the distinctness of A. bunius from its closely
related species. Of the markers tested, psbA-trnH had the highest amplification and RECOMENDATIONS
sequencing success rates as well as parsimony-informative sites and mean
This study supports the amendment of product authentication protocols in
interspecific K2P distances (Table 1).
Table 1. Characteristics of the barcoding loci the Philippines, with the aim of incorporating DNA barcoding in the process
Parameters matK psbA-trnH rbcL of herbal product authentication.
Antidesma sp. 9 9 9
Amplification success 9/9 9/9 9/9
REFERENCES
Sequencing success 7/9 9/9 6/9
Aligned length (bp) 693 558 528
Variable Sites (%) 18 (2.59%) 110 (19.71%) 14 (2.65%)
Parsimony-
Informative Sites (%) 12 (1.73%) 66 (11.83%) 10 (1.89%)

Mean interspecific
0.0076 ± 0.1077 ±
K2P Distances 0.0087 ± 0.0017
0.0008 0.0052
Mean intraspecific
K2P Distances
0.0018 ±
0.0009
0.0438 ±
0.0066
0.0009 ± 0.0012 ACKNOWLEDGEMENTS
Figure 2. SRM of Antidesma spp. UST HERBARIUM