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result(s) for
"Sukrong, Suchada"
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Optimization of Natural Deep Eutectic Solvent-Assisted Extraction of Rosmarinic Acid from Thunbergia laurifolia Lindl. and Evaluation of Antioxidant Activity
by
Thongphichai, Wisuwat
,
Uttarawichien, Tamonwan
,
Khoochonthara, Jasadakorn
in
Acids
,
antioxidant activity
,
Antioxidants
2025
Thunbergia laurifolia Lindl. is a plant known for its promising biological activity, including antioxidant and anti-inflammatory activities, and a rich source of rosmarinic acid (RA). The extraction of T. laurifolia for cosmetic and skincare products using conventional solvents has encountered difficulties, including safety concerns, skin irritation, undesirable odors, and inefficient extraction. In this work, 14 types of natural deep eutectic solvents (NaDESs) with varying compositions and ratios were investigated to compare their efficiency in extracting RA from T. laurifolia by HPLC analysis. The NaDES with the highest extraction efficiency was further utilized in ultrasonic-assisted extraction (UAE), and the extraction parameters were optimized using response surface methodology. The optimized RA content and DPPH scavenging activity were predicted by response surfaces methodology to be 7.52 mg/g DW and 37.6 mg TE/g DW, respectively. The optimal extraction condition was achieved using a propylene glycol-lactic acid NaDES (at an 8:1 molar ratio) with 37% (w/w) H2O, a 30 mL/g liquid-to-solid ratio, an 80 °C extraction temperature, and a 32 min extraction time. The optimized extract was proved to suppress ROS in H2O2-induced keratinocytes. The extract demonstrated robust stability against basic, oxidative, and photolytic stresses, and maintained long-term chemical stability up to 90 days. This study introduces a new green solvent for the effective extraction of T. laurifolia, thereby improving the safety and quality of the extracts for skincare and cosmetic products.
Journal Article
DNA metabarcoding to unravel plant species composition in selected herbal medicines on the National List of Essential Medicines (NLEM) of Thailand
by
Prombutara, Pinidphon
,
Tungphatthong, Chayapol
,
Urumarudappa, Santhosh Kumar J.
in
631/208/514/2254
,
631/449/1659
,
Chloroplasts
2020
Traditional medicines are widely traded across the globe and have received considerable attention in the recent past, with expectations of heightened demand in the future. However, there are increasing global concerns over admixture, which can affect the quality, safety, and efficacy of herbal medicinal products. In this study, we aimed to use DNA metabarcoding to identify 39 Thai herbal products on the Thai National List of Essential Medicines (NLEM) and assess species composition and admixture. Among the products, 24 samples were in-house-prepared formulations, and 15 samples were registered formulations. In our study, DNA metabarcoding analysis using ITS2 and
rbc
L barcode regions were employed to identify herbal ingredients mentioned in the products. The nuclear region, ITS2, was able to identify herbal ingredients in the products at the genus- and family-levels in 55% and 63% of cases, respectively. The chloroplast gene,
rbc
L, enabled genus- and family-level identifications in 58% and 73% of cases, respectively. In addition, plant species were detected in larger numbers (Family identified, absolute %) in registered herbal products than in in-house-prepared formulations. The level of fidelity increases concerns about the reliability of the products. This study highlights that DNA metabarcoding is a useful analytical tool when combined with advanced chemical techniques for the identification of plant species in highly processed, multi-ingredient herbal products.
Journal Article
A PCR-lateral flow immunochromatographic assay (PCR-LFA) for detecting Aristolochia species, the plants responsible for aristolochic acid nephropathy
by
Thongkhao, Kannika
,
Tungphatthong, Chayapol
,
Sukrong, Suchada
in
631/1647/1513/2216
,
631/45/147
,
Acids
2022
Aristolochic acids (AAs), which are strong carcinogens, have caused dietary supplements with
Aristolochia
plants to be discontinued worldwide. Therefore, the development of a method to identify these herbs is critical for customer safety. To support the regulation of
Aristolochia
-free products, a PCR coupled with lateral flow immunochromatographic assay (PCR-LFA) that is specific to the nucleotide signature in plastid
rbc
L gene region of
Aristolochia
species was developed to detect
Aristolochia
plants and related herbal products. Triplex primers (A397F, C357F and R502) were designed based on specific nucleotides observed exclusively in the
rbc
L sequences of
Aristolochia
. Positive results for
Aristolochia
occur when the three pink lines are clearly developed on the developed lateral flow strip and can be seen by the naked eye. In this study, the lateral flow strip has sensitivity for detecting amplicons amplified from genomic DNA at the concentrations as low as 0.01 ng. Various kinds of samples, including purchased crude drugs and polyherbal samples, have been investigated, and the results showed that
Aristolochia
crude drugs and
Aristolochia
-containing products are still present in dispensaries. In conclusion, with the goal of protecting consumers from the health risks associated with
Aristolochia
contamination, PCR-LFA was developed and demonstrated to be efficient for detecting plants belonging to
Aristolochia
in various kinds of samples.
Journal Article
DNA barcoding of Aristolochia plants and development of species-specific multiplex PCR to aid HPTLC in ascertainment of Aristolochia herbal materials
by
Sukrong, Suchada
,
Aumnouypol, Surattana
,
Denduangboripant, Jessada
in
Admixtures
,
Aristolochia
,
Aristolochia - classification
2018
The anecdotal evidence is outstanding on the uses of Aristolochia plants as traditional medicines and dietary supplements in many regions of the world. However, herbal materials derived from Aristolochia species have been identified as potent human carcinogens since the first case of severe renal disease after ingesting these herbal preparations. Any products containing Aristolochia species have thus been banned on many continents, including Europe, America and Asia. Therefore, the development of a method to identify these herbs is critically needed for customer safety. The present study evaluated DNA barcoding of the rbcL, matK, ITS2 and trnH-psbA regions among eleven Aristolochia species collected in Thailand. Polymorphic sites were observed in all four DNA loci. Among those eleven Aristolochia species, three species (A. pierrei, A. tagala and A. pothieri) are used as herbal materials in Thai folk medicine, namely, in Thai \"Krai-Krue\". \"Krai-Krue\" herbs are interchangeably used as an admixture in Thai traditional remedies without specific knowledge of their identities. A species-specific multiplex PCR based on nucleotide polymorphisms in the ITS2 region was developed as an identification tool to differentiate these three Aristolochia species and to supplement the HPTLC pattern in clarifying the origins of herbal materials. The combination of multiplex PCR and HPTLC profiling achieves accurate herbal identification with the goal of protecting consumers from the health risks associated with product substitution and contamination.
Journal Article
Differentiation of Mitragyna speciosa, a narcotic plant, from allied Mitragyna species using DNA barcoding-high-resolution melting (Bar-HRM) analysis
by
Sooksawate, Thongchai
,
Tungphatthong, Chayapol
,
Awachai, Supita
in
631/449/1659
,
631/61
,
631/61/447
2021
Mitragyna speciosa
(Korth.) Havil. [MS], or “kratom” in Thai, is the only narcotic species among the four species of
Mitragyna
in Thailand, which also include
Mitragyna diversifolia
(Wall. ex G. Don) Havil. [MD],
Mitragyna hirsuta
Havil. [MH], and
Mitragyna rotundifolia
(Roxb.) O. Kuntze [MR].
M. speciosa
is a tropical tree belonging to the Rubiaceae family and has been prohibited by law in Thailand. However, it has been extensively covered in national and international news, as its abuse has become more popular.
M. speciosa
is a narcotic plant and has been used as an opium substitute and traditionally used for the treatment of chronic pain and various illnesses
.
Due to morphological disparities in the genus, the identification of plants in various forms, including fresh leaves, dried leaf powder, and finished products, is difficult. In this study, DNA barcoding combined with high-resolution melting (Bar-HRM) analysis was performed to differentiate
M. speciosa
from allied
Mitragyna
and to assess the capability of Bar-HRM assays to identify
M. speciosa
in suspected kratom or
M. speciosa
-containing samples. Bar-HRM analysis of PCR amplicons was based on the ITS2,
rbc
L,
trn
H
-psb
A
,
and
mat
K DNA barcode regions. The melting profiles of ITS2 amplicons were clearly distinct, which enabled the authentication and differentiation of
Mitragyna
species from allied species. This study reveals that DNA barcoding coupled with HRM is an efficient tool with which to identify
M. speciosa
and
M. speciosa
-containing samples and ensure the safety and quality of traditional Thai herbal medicines.
Journal Article
Automated home-cage-like monitoring for assessing innate behaviors in a murine hangover model
by
Nwe, San Yoon
,
Sukrong, Suchada
,
Towiwat, Pasarapa
in
692/1807/1693
,
692/308/1426
,
692/308/2778
2025
The prevalence of alcohol consumption among the younger generation remains alarmingly high. A hangover is a common short-term consequence observed after consuming alcohol. To effectively study alcohol-induced hangovers, reliable and translational animal models, along with appropriate testing methods, are required. While several testing approaches have been used in hangover-induced mice, they often fail to assess innate behaviors comprehensively and are limited by short observation periods. Although existing studies have developed methods to assess hangover-related behaviors in rodents, few have focused on innate behaviors. This study aimed to establish a model for assessing the innate behaviors of hangover-induced mice using automated home-cage-like behavioral monitoring. Mice were intraperitoneally injected with ethanol at doses of 3, 2, or 1 g/kg, followed by behavioral assessments, including exploratory actions and long-term home-cage-like behaviors during both day and night phases. Results showed a significant reduction in mobile behaviors (climbing, locomotion, rearing), speed, and distance traveled, along with increased immobility in both exploratory and long-term home-cage-like assessments. Furthermore, there was a significant decrease in exploratory behaviors and long-term home-cage-like activities, which were linked to hangover symptoms. This study provides a preliminary approach for assessing hangover behaviors in mice using automated behavioral monitoring, ensuring improved animal welfare, optimised timing, and extended assessment durations. Hence, we propose automated home-cage-like behavioral assessment as an exploratory model for evaluating hangover behaviors in mice, which may serve as a useful tool for future research on the therapeutic efficacy of anti-hangover compounds.
Journal Article
Exploring the safety of lycorine in the central nervous system and its impact on pain-like behaviors in mice
by
Thongphichai, Wisuwat
,
Wasana, Peththa Wadu Dasuni
,
Sukrong, Suchada
in
631/154/1438
,
631/154/436
,
Alkaloid
2024
Alkaloid analgesics have been associated with adverse effects on the central nervous system (CNS). Therefore, it is crucial to characterize the effects of alkaloid analgesics. Plants rich in lycorine, an alkaloid, have shown promise as analgesics. However, the exploration of their CNS side effects, and analgesic effectiveness remains incomplete. The aim of the present study was to investigate the CNS safety profiles of lycorine and its potential analgesic efficacy. Lycorine (3, 10, and 30 mg/kg, intraperitoneal) did not affect motor coordination, and doses of 3 and 10 mg/kg of lycorine did not lead to any impairment in spontaneous locomotor activity. However, the highest dose (30 mg/kg) demonstrated a significant impairment in rearing behavior and an increase in immobility. The safety doses were subsequently used to assess the analgesic efficacy of lycorine in a mouse model of inflammatory pain. Lycorine (1, 3, and 10 mg/kg, intraperitoneal) demonstrated a dose-dependent reduction in pain-like behaviors in formalin-induced mice. In the in vitro study, lycorine regulated immune cells, suggesting its involvement as a cellular mechanism underlying the suppression of pain-like behaviors observed in the formalin model. Overall, our findings delineate the CNS safety range of lycorine in mice and suggest its potential use as an analgesic.
Journal Article
Combining DNA and HPTLC profiles to differentiate a pain relief herb, Mallotus repandus, from plants sharing the same common name, “Kho-Khlan”
by
Pichetkun, Vipawee
,
Gaewtongliam, Suthathip
,
Thongkhao, Kannika
in
Analysis
,
Bar codes
,
Biology and life sciences
2022
The pain relief formula “Ya Pa Som Kho-Khlan (YPSKK)” or “ยาผสมโคคลาน” in Thai is officially recorded in the Natural List of Essential Medicines (NLEM) of Thailand. The main component is Mallotus repandus (Willd.) Müll. Arg.; however, Anamirta cocculus (L.) Wight & Arn and Croton caudatus Gleiseler share the same common name: “Kho-Khlan”. Confused usage of A . cocculus or C . caudatus can have effects via toxicity or unsuccessful treatment. This study aimed to combine a high-performance thin-layer chromatography (HPTLC) technique and DNA barcoding coupled with high-resolution melting (Bar-HRM) to differentiate M . repandus from the other two species. The M . repandus extract exhibited a distinct HPTLC profile that could be used to differentiate it from the others. DNA barcodes of the rbc L, mat K, ITS and psb A- trn H intergenic spacer regions of all the plants were established to assist HPTLC analysis. The rbc L region was selected for Bar-HRM analysis. PCR amplification was performed to obtain 102 bp amplicons encompassing nine polymorphic nucleotides. The amplicons were subjected to HRM analysis to obtain melting curve profiles. The melting temperatures (T m ) of authentic A . cocculus (A), C . caudatus (C) and M . repandus (M) were separated at 82.03±0.09°C, 80.93±0.04°C and 80.05±0.07°C, respectively. The protocol was applied to test crude drugs (CD1-6). The HPTLC profiles of CD2-6 showed distinct bands of M . repandus , while CD1 showed unclear band results. The Bar-HRM method was applied to assist the HPTLC and indicated that CD1 was C . caudatus . While ambiguous melting curves from the laboratory-made formulae were obtained, HPTLC analysis helped reveal distinct patterns for the identification of the plant species. The combination of HPTLC and Bar-HRM analysis could be a tool for confirming the identities of plant species sharing the same name, especially for those whose sources are multiple and difficult to identify by either chemical or DNA techniques.
Journal Article
Integrating untargeted volatile metabolomics and molecular evidence supporting chemotaxonomy in Kaempferia species for more effective identification
by
Wongsuwan, Pornpimon
,
Phokham, Boonmee
,
Rattanakrajang, Pantamith
in
631/181
,
631/181/2480
,
631/181/757
2025
Kaempferia
L., a medicinal genus of Zingiberaceae family, is widely distributed from India to Southeast Asia and is rich in terpenoids, flavonoids, phenolics, and volatile oils. Recently, it has gained attention for its diverse biological activities, including antioxidant, anticancer, analgesic, anti-inflammatory, and anti-tuberculosis effects. However, several
Kaempferia
species complexes exhibit similar morphological characteristics, making identification and classification challenging. This study integrates morphology, molecular phylogeny, and phytochemistry to identify and distinguish
Kaempferia
species. Phylogenetic relationships were reconstructed using four DNA barcoding markers: one nuclear region (ITS) and three chloroplast markers (
mat
K,
rbc
L, and
psb
A-
trn
H). Untargeted metabolomic analysis using SPME-GC-MS, combined with multivariate statistical analyses, was employed to resolve species relationships and display volatile profiles among 15
Kaempferia
species from two subgenera. A total of 217 metabolites were identified by the SPME-GC-MS technique. Variable Importance in Projection (VIP ≥ 1.5) analysis indicated 30 key metabolites, primarily sesquiterpenes, as specific chemotaxonomic markers. This study provides a comprehensive chemical profile of
Kaempferia
species and highlights metabolomic differences among them. Our findings emphasize the importance of integrating morphological, molecular, and phytochemical approaches for precise identification of closely related species, particularly within
Kaempferia
. This chemotaxonomic research also provides further applications for species authentication in pharmaceuticals and medicine.
Journal Article
Anticancer Effects and Molecular Action of 7-α-Hydroxyfrullanolide in G2/M-Phase Arrest and Apoptosis in Triple Negative Breast Cancer Cells
by
Graidist, Potchanapond
,
Srisawat, Theera
,
Sukrong, Suchada
in
7-α-hydroxyfrullanolide
,
Adapter proteins
,
anticancer activity
2022
Triple negative breast cancer (TNBC) is a breast cancer subtype characterized by the absence of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 expression. TNBC cells respond poorly to targeted chemotherapies currently in use and the mortality rate of TNBC remains high. Therefore, it is necessary to identify new chemotherapeutic agents for TNBC. In this study, the anti-cancer effects of 7-α-hydroxyfrullanolide (7HF), derived from Grangea maderaspatana, on MCF-7, MDA-MB-231 and MDA-MB-468 breast cancer cells were assessed using MTT assay. The mode of action of 7HF in TNBC cells treated with 6, 12 and 24 µM of 7HF was determined by flow cytometry and propidium iodide (PI) staining for cell cycle analysis and annexin V/fluorescein isothiocyanate + PI staining for detecting apoptosis. The molecular mechanism of action of 7HF in TNBC cells was investigated by evaluating protein expression using proteomic techniques and western blotting. Subsequently, 7HF exhibited the strongest anti-TNBC activity toward MDA-MB-468 cells and a concomitantly weak toxicity toward normal breast cells. The molecular mechanism of action of low-dose 7HF in TNBC cells primarily involved G2/M-phase arrest through upregulation of the expression of Bub3, cyclin B1, phosphorylated Cdk1 (Tyr 15) and p53-independent p21. Contrastingly, the upregulation of PP2A-A subunit expression may have modulated the suppression of various cell survival proteins such as p-Akt (Ser 473), FoxO3a and β-catenin. The concurrent apoptotic effect of 7HF on the treated cells was mediated via both intrinsic and extrinsic modes through the upregulation of Bax and active cleaved caspase-7–9 expression and downregulation of Bcl-2 and full-length caspase-7–9 expression. Notably, the proteomic approach revealed the upregulation of the expression of pivotal protein clusters associated with G1/S-phase arrest, G2/M-phase transition and apoptosis. Thus, 7HF exhibits promising anti-TNBC activity and at a low dose, it modulates signal transduction associated with G2/M-phase arrest and apoptosis.
Journal Article