Simultaneous quantification of chlorogenic acid, quercetin, and kaempferol in Urceola rosea leaves by CE and HPLC techniques: method validation and comparative study.

Two analytical methods, CE and HPLC, were developed, validated, and compared for simultaneously determining chlorogenic acid, quercetin, and kaempferol in Urceola rosea leaves collected from twelve different sources of Thailand. The determination was accomplished by CE on the uncoated fused silica capillary tube and by HPLC on the ZORBAX Eclipse Plus C18 column. Chlorogenic acid was found to have the highest average amount in U. rosea leaves, followed by kaempferol and quercetin, respectively. Compared to CE, HPLC is considered to provide higher sensitivity and robustness as indicated by lower LOD and LOQ values as well as % RSD of robustness for all analytes. Overall, the present study first demonstrated that both methods were selective, sensitive, and robust, allowing reliable simultaneous quantification of three major bioactive compounds in U. rosea leaves, which could be beneficially applied for quality control assessment of this plant material, particularly those cultivated in Thailand.

Polyherbal formulation exerts wound healing, anti-inflammatory, angiogenic and antimicrobial properties: Potential role in the treatment of diabetic foot ulcers.

Diabetic foot ulcer (DFU) is a common and devastating complication in diabetic patients and is associated with an elevated risk of amputation and mortality. DFU remains a major therapeutic challenge due to poor understanding of its underlying pathogenesis. This complication is characterized by impaired wound healing; however, mechanisms causing this impairment are complicated and involve interactions between many different cell types and infections. In addition to other conventional DFU treatments, herbal foot baths are also common, although little is known about their mechanisms of action, and they contain a wide variety of herbal ingredients. In this study, we aimed to examine the effects of three polyherbal formulations consisting of medicinal plants used in traditional Thai herbal foot baths on wound healing, anti-inflammation, angiogenesis, and extracellular matrix modulation using high-concentration glucose-treated human keratinocytes, in addition to antibacterial evaluation. Our results showed that formulation 3 (F3) possessed the greatest potential to restore the impairment of keratinocytes caused by high glucose concentrations. We found that F3 could inhibit the growth of Staphylococcus aureus, accelerate wound healing, and upregulate the expression of TIMP-1, VEGF, and TGF-ß, and downregulate the expression of TNF-α, IL-6, and MMP-9. Collectively, these data support the potential of F3 for therapeutic development in the treatment of DFU.

Standardization of Leonurus sibiricus L. aerial part and capillary electrophoresis quantitative analysis of its leonurine content.

The quality parameters of Leonurus sibiricus L. aerial part crude drugs were evaluated. Fifteen crude drugs were collected from various locations throughout Thailand. The transverse section of the stem of L. sibiricus showed quadrangular character highlighted the ribs with angular collenchyma. The epidermis was uniseriate with abundant glandular trichomes distribution. Prismatic calcium oxalate prisms were found in the stem medullary parenchyma.The histological character of crude drug powder showed bordered pitted vessel, fragment of fiber, glandular trichome, prism crystal, spiral vessel, starch granule, and stomata. The loss on drying, total ash, acid-insoluble ash, and moisture contents should be not more than 8.18, 15.28, 4.04, and 8.91 g/100 g dry weight, whereas ethanol and water-soluble extractive values should be not less than 7.67, and 17.21 g/100 g of dry weight, respectively. Leonurine in the crude drugs were analyzed by capillary electrophoresis (CE) with photodiode array detector. The ethanolic extraction performed by Soxhlet apparatus yielded 18.86 ± 4.09 g/100 g dry weight. The electropherogram detected at 277 nm showed the migration time of leonurine at 6.2 min. The developed CE was found to be valid for leonurine quantification in L. sibiricus ethanolic extract. The contents of leonurine in 15 crude drugs ranged from 0.79 to 4.23 mg/g with the average of 2.38 ± 1.10 mg/g dry weight. This study established the pharmacognostic specification of L. sibiricus crude drug in Thailand with special reference to a bioactive compound, leonurine. CE was beneficial technique for the analysis of leonurine in L. sibiricus aerial parts.

In vitro cytotoxic, genotoxic, and antityrosinase activities of Clitoria macrophylla root.

Clitoria macrophylla Wall. (Leguminosae), locally known as Non-tai-yak or An-chan-pa, commonly distributed in tropical nations and Southeast Asia. Regarding traditional Thai medical system, C. macrophylla roots carry out a potential in dermatology. Its roots are also used as insecticide in agriculture and animal farming. Moreover, clitoriacetal is the major component that can be detected in C. macrophylla root. This research aimed to assess the efficacy of C. macrophylla root extract and clitoriacetal for its anticancer and antityrosinase activities as well as to assess in vitro safety potential for its cytotoxic and genotoxic effects. C. macrophylla root and clitoriacetal were tested by brine shrimp lethality, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, comet assay, and antityrosinase activity. C. macrophylla root, clitoriacetal, and rotenone demonstrated the toxicity against brine shrimp nauplii with LC50 of 332.15, 136.54, and 0.15 µg/mL, respectively. C. macrophylla root and clitoriacetal showed cytotoxic potential against breast ductal carcinoma (BT-474), liver hepatoblastoma (Hep-G2), and colon adenocarcinoma (SW-620). At 100 µg/mL, the percent DNA damage of C. macrophylla root and clitoriacetal was 37.84% and 36.01%, respectively. C. macrophylla root and clitoriacetal were able to inhibit the tyrosinase enzyme with IC50 of 12.27 and 7.30 mg/mL, respectively, which less effective than glutathione (positive control). The present study revealed the in vitro biological activities of C. macrophylla root and its clitoriacetal constituent which proposed the scientific evidences in efficacy and safety evaluation including in vitro cytotoxicity, DNA damage as well as antityrosinase activities.

Quality evaluation with reference to clitoriacetal and in vitro antioxidant activities of Clitoria macrophylla root.

Clitoria macrophylla Wall. (syn. Clitoria hanceana Hemsl.), is commonly known in Thai as Nontai-yak, An-chan-pa, or Ueang-chan-pa, which belongs to Leguminosae family. According to traditional Thai medicine, the root has been used for the treatment of skin diseased as well as for pest control in horticulture and animal husbandry. The aim of this study is to investigate for the pharmacognostic specification, the clitoriacetal content, and in vitro antioxidant capacities of C. macrophylla roots from 12 different sources throughout Thailand. Clitoriacetal content was quantitatively analyzed by thin-layer chromatography (TLC) - densitometry with winCATS software and TLC image analysis with ImageJ software. Antioxidant activities were evaluated by 2, 2-diphenyl-1-picrylhydrazyl radical scavenging, ferric reducing antioxidant power assay, ß-carotene bleaching assay, total phenolic, and total flavonoid contents. The pharmacognostic specification of C. macrophylla roots in Thailand was established. The loss on drying, total ash, acid-insoluble ash, and water contents should be not more than 6.40%, 12.29%, 8.89%, and 8.16% of dry weight, whereas ethanol and water-soluble extractive values should be not less than 4.95% and 14.72% of dry weight, respectively. Furthermore, the clitoriacetal content of C. macrophylla roots determined by TLC-densitometry and TLC image analysis was found to be 2.20 ± 1.31 and 2.22 ± 1.16 g/100 g of dry roots, respectively. The clitoriacetal contents of both methods were not significantly different using paired t-test. Moreover, the ethanolic extract of C. macrophylla roots showed its antioxidant potential compared to the standard butylated hydroxyl toluene and quercetin.

A new approach for the detection of cervical cancer in Thai women.

OBJECTIVES: The objective was to screen cervical cell samples of Thai women by using the Fourier-transform infrared (FTIR) spectrophotometry technique; the results were compared to the histologic diagnosis (gold standard). METHODS: FTIR spectrophotometry is a new technology for cervical cancer screening. Instead of detecting the morphological changes as used in Pap smear test, this technique detects, at the molecular level, structural changes of functional groups through the changes of the infrared absorption spectrum. When the infrared light is passed through a cervical cell sample, a molecule absorbs infrared radiation of the appropriate frequency which excites it from one vibrational or rotational level to another. A graph of energy absorbed versus frequency is the absorption spectrum of the sample. The FTIR spectra can be interpreted as normal and abnormal results. First, significant changes in the intensity ratios and, second, significant shifts of the peak frequencies were detected. Two hundred seventy-five cervical cell specimens were received from patients undergoing hysterectomy at the Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University Hospital. Samples were collected, prepared, and analyzed by FTIR spectroscopy. RESULTS: Histological examinations showed 108 abnormal cases and 167 normal cases. FTIR results versus histology showed sensitivity of 96.3% and specificity of 96.4%. False-negative and false-positive rates were 3.7 and 3.6%, respectively. CONCLUSIONS: FTIR showed high sensitivity and yielded a good false-negative rate. Besides the cervical cancer detection, the FTIR spectroscopy technique can also elicit positive results from adenocarcinoma of the endometrium, sarcoma of the uterus, and ovarian malignancies.