Identification of Apigenin and Luteolin in Artemisia annua L. for the Quality Control

Objective: To identify active compounds and establish the chemical fingerprint of Artemisia annua L. for the quality control. Methods: Thin-layer chromatography (TLC) conditions were developed to screen for 2 common flavonoids (apigenin and luteolin). Three mobile phases were used to isolate these flavonoids in 80% ethanolic extract of A. annua. Hexane : ethyl acetate : acetic acid (31:14:5, v/v) and toluene : 1,4-dioxane : acetic acid (90:25:4, v/v) were used in normal phase TLC (NP-TLC), and 5.5% formic acid in water : methanol (50:50, v/v) were used in reverse phase TLC (RP-TLC). Chromatograms were visualized under visible light after spraying with Fast Blue B Salt. Apigenin and luteolin bands were checked by comparing their Rf values and UV-Vis absorption spectra with reference markers. Results: Apigenin and luteolin were simultaneously detected with good specificity in RP-TLC condition, while only apigenin was detected in NP-TLC condition. Apigenin band intensity was higher than luteolin band intensity in both conditions. Conclusion: This knowledge can be applied to the development of quality control assessments to ensure product efficacy and consistency

Identification of Apigenin and Luteolin in Artemisia annua L. for the Quality Control

Objective: To identify active compounds and establish the chemical fingerprint of Artemisia annua L. for the quality control. Methods: Thin-layer chromatography (TLC) conditions were developed to screen for 2 common flavonoids (apigenin and luteolin). Three mobile phases were used to isolate these flavonoids in 80% ethanolic extract of A. annua.Hexane : ethyl acetate : acetic acid (31:14:5, v/v) and toluene : 1,4-dioxane : acetic acid (90:25:4, v/v) were used in normal phase TLC (NP-TLC), and 5.5% formic acid in water : methanol (50:50, v/v) were used in reverse phase TLC (RP-TLC). Chromatograms were visualized under visible light after spraying with Fast Blue B Salt. Apigenin and luteolin bands were checked by comparing their Rf values and UV-Vis absorption spectra with reference markers. Results: Apigenin and luteolin were simultaneously detected with good specificity in RP-TLC condition, while only apigenin was detected in NP-TLC condition. Apigenin band intensity was higher than luteolin band intensity in both conditions. Conclusion: This knowledge can be applied to the development of quality control assessments to ensure product efficacy and consistenc

The Effects of Thai Herbal Ha-Rak Formula on COX Isoform Expression in Human Umbilical Vein Endothelial Cells Induced by IL-1 β

Objective. To investigate the modulated effects of HRF on cyclooxygenase isoform expression and its activity, using the human umbilical vein endothelial cell (HUVEC) model induced by interleukin-1 beta (IL-1β). Methods. Cells were treated with indomethacin (positive control), HRF, and its components at various concentrations prior to treatment with IL-1β at 24 h. Cell viability was determined by MTT assay. Moreover, the anti-inflammatory effects of HRF and its components through mRNA and protein expression were established using real-time quantitative PCR and Western blot, respectively. COX activity was identified via exogenous and endogenous PGE2 productions using the EIA. Result. There was no cytotoxicity in HUVECs treated with HRF. None of the experimental conditions used in the study affected the expression of COX-1, but COX-2 protein expression was inhibited at concentrations under 10 µg/mL. Despite the significantly increased levels of exogenous PGE2, HRF had no effect on COX-2 mRNA expression. However, the production of PGE2 was lower at a concentration of 100 µg/mL HRF than at a concentration below 10 µg/mL. Interestingly, each component of HRF revealed different effects of the Ha-Rak formula. Conclusion. Our preliminary findings suggest that HRF and its components provide diverse modulation of COX-2 and PGE2 at the in vitro level

Study the Effect of an Antipyretic Drug, Thai Herbal Ha-Rak Formula on Platelet Aggregation in Healthy Thai Volunteers: A Randomized, Placebo - Controlled Trial

Background: Fever may alter platelet homeostasis leading to bleeding tendency. Thai herbal Ha-rak formula (HRF), a traditional Thai remedy consisting of five medicinal plants, is indicated for relieving fever. However, its effect on platelet has not been evaluated. This study aims to investigate the effect of HRF on platelet aggregation. Methods: A randomized, placebo, controlled trial was carried out in 46-healthy Thai volunteers, both male and female. The subjects either received the maximum recommended dose per day of HRF (1,500 mg) or placebo. Platelet aggregation, using aggregometer (AggRam), was assessed in platelet rich plasma (PRP) in response to each of three different agonists including epinephrine, adenosine diphosphate (ADP) and collagen at pre-dose and 8, 32 hours and 7-10 days after the first dose. Results: All participants completed the study. Only few adverse events occurred which spontaneously improved without further treatment. Overall, analysis of platelet activity compared before and after HRF administration did not show significant difference of maximum percentage of platelet aggregation at any time point except the platelet response to collagen at 32 hours and 1 week after the first HRF dose. However, subgroup analysis characterized by sex, and platelet aggregation in response to all agonists did not reveal any significant change. The same results applied to subgroup analysis based on the different patterns of platelet aggregation. Conclusion: HRF at a dose of 1,500 mg/day is well tolerated and has a significant effect on platelet aggregation only when induced by collagen

Redox Mechanisms of AVS022, an Oriental Polyherbal Formula, and Its Component Herbs in Protection against Induction of Matrix Metalloproteinase-1 in UVA-Irradiated Keratinocyte HaCaT Cells

Ayurved Siriraj HaRak (AVS022) formula has been used for topical remedy of dermatologic disorders. Oxidative stress induced by ultraviolet (UV) A irradiation could be implicated in photoaged skin through triggering matrix metalloproteinase-1 (MMP-1). We, therefore, explored the antioxidant mechanisms by which AVS022 formulation and its individual components protected against UVA-dependent MMP-1 upregulation in keratinocyte HaCaT cells. TLC analysis revealed the presence of multiple phenolics including gallic acid (GA) in the AVS022 extracts. We demonstrated that pretreatment with the whole formula and individual herbal components except T. triandra protected against increased MMP-1 activity in irradiated HaCaT cells. Moreover, all herbal extracts and GA, used as the reference compound, were able to reverse cytotoxicity, oxidant production, glutathione (GSH) loss, and inactivation of catalase and glutathione peroxidase (GPx). F. racemosa was observed to yield the strongest abilities to abolish UVA-mediated induction of MMP-1 and impairment of antioxidant defenses including GSH and catalase. Our observations suggest that upregulation of endogenous antioxidants could be the mechanisms by which AVS022 and its herbal components suppressed UVA-stimulated MMP-1 in HaCaT cells. In addition, pharmacological actions of AVS022 formula may be attributed to the antioxidant potential of its components, in particular F. racemosa, and several phenolics including GA

The Effect of Ayurved Siriraj Wattana Recipe (AVS073) on LPS Induced COX-2 Expression in Human PBMC

Objective: To investigate the anti-inflammatory effects of AVS073 via the expression of COX-2 in human peripheral blood mononuclear cells (PBMC) induced by LPS. Methods: PBMC from healthy volunteers were treated with LPS alone, AVS073 alone (0.1-10 μg/ml) and LPS plus AVS073 for 18 hours. Cell proliferation was measured using MTT assay, while COX-2 mRNA, COX-2 protein and PGE2 production were assessed using real time RT-PCR, western blot and enzyme immunoassay (EIA), respectively. Results: It was found that neither LPS nor AVS073 has effects on cell proliferation. The optimum time of LPS- induced COX-2 expression was 18 hours. Even the expression of COX-2 mRNA showed trends toward decrease in all concentrations of AVS073 treatment, it was not significantly different. However, the expression of COX-2 protein was significantly decreased by 10 μg/ml AVS073. Surprisingly, endogenous PGE2 was significantly increased, whereas exogenous PGE2 was not significantly changed. Conclusion: AVS073 has some anti-inflammatory effects, probably at the level of post translational modification. Increase in endogenous PGE2 production, in spite of the decrease in the expression of COX-2 protein, may be due to the other factors or isoforms of COX

Additional file 1: of IL-1β-induced modulation of gene expression profile in human dermal fibroblasts: the effects of Thai herbal Sahatsatara formula, piperine and gallic acid possessing antioxidant properties

The overview of microarray analysis work flow (Figure S1) and the list of human housekeeping genes matching with gene array in microarray BeadChip (Table S1). (DOCX 32 kb

Chemical Profiling of an Antipyretic Drug, Thai Herbal Harak Formula, by Liquid Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry

Objective: To isolate and identify chemical compounds in Harak, a Thai herbal formula widely used as an antipyretic drug in Thailand. Methods: Methanol extraction of the Harak formula and its five herbal components were separated by Ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. This method has been developed for “untargeted” profiling of this formula and its components. An online database was used to identify chemical compounds by comparing its empirical molecular formula, isotope pattern, and fragmentation pattern. Results: Nineteen chemical constituents were found from positive and negative electrospray ionization (ESI) modes. These compounds included flavonoids, hesperitine, and iso-corydine, which are known to possess antioxidant and anticancer activity. Moreover, data from the principle component analysis (PCA) score plot of positive and negative ESI modes showed that the chemical constituents of Thai herbal Harak formula were similar to those found in Ficus racemosa Linn. and Capparis micracantha DC. Conclusion: Under this optimization method, nineteen chemical constituents including phenolic and flavonoids were characterized in both positive and negative ESI mode

The Efficacy of Thai Herbal Prasaplai Formula for Treatment of Primary Dysmenorrhea: A Short-Term Randomized Controlled Trial

This study aims to compare the efficacy of pain relief between a specific Thai herbal Prasaplai formula (PPF) and placebo in patients with primary dysmenorrhea. Forty women with primary dysmenorrhea symptoms were randomized into two groups. The experimental group received PPF capsules 1000 mg orally three times per day before meals for three days starting from the first day of menstruation. The placebo group received placebo as the same dose and time. Average pain intensity from the first day to the third day of cycle significantly decreased in both groups ( < 0.001), but with no statistically significant difference between groups. Using a pre-and posttreatment difference in NRS of at least 2, a greater proportion of patients in PPF group experienced pain relief compared to placebo during the first and second day of period. A greater proportion of PPF group also experienced no pain compared to the placebo group on day 1 and day 3 by using multidimensional scoring. The PPF demonstrates pain relief activity when used during menstruation in primary dysmenorrhea patients, with no adverse effects. However, further studies are needed in order to assess the value of PPF as a long-term prophylaxis for primary dysmenorrhea