Impact of Tea Processing on Tryptophan, Melatonin, Phenolic and Flavonoid Contents in Mulberry (Morus alba L.) Leaves: Quantitative Analysis by LC-MS/MS.

Mulberry (Morus alba L.) leaves from two cultivars, Yai-Burirum (YB) and Khunphai (KP), were prepared into green tea (GT) and black tea (BT). Compared to fresh leaf (FL) extract, GT and BT extracts were evaluated for their total phenolic and total flavonoid contents. Total phenolic content (TPCs) in all samples ranged between 129.93 and 390.89 mg GAE/g extract. The processing of tea decreased the levels of TPC when compared to FL extracts in both cultivars. The total flavonoid content (TFCs) in all samples was found in the range of 10.15-39.09 mg QE/g extract and TFCs in GT and BT extracts were higher than FL extracts. The change in tryptophan, melatonin, phenolic and flavonoid contents was investigated by liquid chromatography-mass spectroscopy (LC-MS). The results exhibited that tryptophan contents in all samples were detected in the range 29.54-673.72 µg/g extract. Both GT and BT extracts increased tryptophan content compared to FL extracts. BT extracts presented the highest amounts of tryptophan among others in both cultivars. Phenolic compounds were found in mulberry leaf extracts, including gallic acid, caffeic acid, gentisic acid, protocatechuic acid and chlorogenic acid. Chlorogenic acid presented the highest amount in all samples. Almost all phenolic acids were increased in the processed tea extracts except chlorogenic acid. Rutin was the only flavonoid that was detected in all extracts in the range 109.48-1009.75 mg/g extract. The change in phenolic and flavonoid compounds during tea processing resulted in the change in antioxidant capacities of the GT and BT extracts. All extracts presented acetylcholinesterase enzyme (AChE) inhibitory activity with IC50 in the range 146.53-165.24 µg/mL. The processing of tea slightly increased the AChE inhibitory effect of GT and BT extracts. In conclusion, processed tea from mulberry leaves could serve as a new alternative functional food for health-concerned consumers because it could be a promising source of tryptophan, phenolics and flavonoids. Moreover, the tea extracts also had antioxidative and anti-AChE activities.

Comparative Effectiveness of Thai Herbal Formula (Thor-Ra-Nee-San-Tha-Kat) Versus Naproxen for Chronic Myofascial Pain: A Pilot Randomized-Controlled Trial.

Background: Myofascial pain syndrome (MPS) is a painful musculoskeletal condition. The prevalence of MPS ranges from 5.9% to 38.7% in the general population. "Thor-ra-nee-san-tha-kat" (TRK) is a traditional formula included in the Thailand National List of Essential Medicines for the treatment for muscle pain caused by abdominal rigidity and for severe constipation. Objectives: The authors employed a pilot single-blind, randomized-controlled trial to compare the effectiveness of TRK and naproxen for the treatment of chronic upper trapezius MPS. Materials and Methods: Seventy-six male and female subjects, ages 25-55 years, who met the inclusion criteria were equally randomized into two groups to receive either two 500 mg capsules of TRK once daily before bed or two 250 mg naproxen tablets twice a day after meals for 14 days. Subjects assessed their level of pain using the numerical rating scale. Cervical range of motion (CROM) was determined using a goniometer, and pressure pain threshold (PPT) was assessed using an algometer. Adverse drug reactions were recorded and all items were compared within and between groups, before and after treatment. Results: The results revealed that patient pain scores after 14 days of treatment were much improved with mean differences exceeding the reference minimum clinically important difference (MCID) in both groups. However, the changes in CROM and PPT values were small and did not surpass their respective reference MCIDs except for the right lateral bending CROM for naproxen treatment. The adverse drug reactions were mild, with watery stools reported by 47% of patients in the TRK-treated group and constipation reported by 24% of those in the naproxen group. Conclusion: The administration of TRK formula for 14 days was safe and as effective as naproxen at providing short-term relief of pain in patients with chronic upper trapezius pain.

Determination of melatonin content in traditional Thai herbal remedies used as sleeping aids.

BACKGROUND: Melatonin content was screened in leaves of seven edible herbs used as sleeping aids in Thai traditional medicine. These plants are Piper nigrum L, Sesbania glandiflora (L.) Desv., Sesbania sesban (L.) Merr., Senna tora (L.) Roxb., Moringa oleifera Lam., Momordica charantia L. and Baccaurea ramiflora Lour. Dried leaves were extracted by sonication in methanol for six hours at room temperature, and then melatonin was purified by C18 solid phase extraction (SPE). Melatonin was then quantified by a validated RP-C18 HPLC method with fluorescent detection. FINDINGS: Melatonin contents in extracts of B. ramiflora, S. glandiflora, M. charantia, S. tora and S. sesban were 43.2, 26.3, 21.4, 10.5 and 8.7 ng/g of dry sample weight, respectively. The highest melatonin content was from P. nigrum extract (1092.7 ng/g of dry sample weight). Melatonin was not detected in the extract of M. oleifera. Melatonin identification was confirmed by ELISA. CONCLUSIONS: Melatonin was found in six of the seven herbs in the traditional Thai sleeping recipe. One of these, P. nigrum, exhibited an encouragingly high amount of melatonin.

Quantitative analysis of barakol content in Senna siamea leaves and flowers by TLC-densitometry.

OBJECTIVE: To develop a TLC-densitometric method for the determination of barakol content in Senna siamea leaf and flower extracts, and to compare the barakol content in mature leaves, young leaves and young flowers of the plant which are consumed as a vegetable in curry. MATERIALS AND METHODS: The extraction of pure barakol was performed by boiling the fresh young leaves of S. siamea with 0.5% sulfuric acid followed by chloroform extraction. The extract was further purified and recrystallized from absolute ethanol. Authentic sample of barakol was used for the validation of the TLC-densitometric method. Chromatography was performed on a TLC aluminium plate precoated with silica gel 60 F(254)as a stationary phase and chloroform-methanol (85:15 v/v) as a solvent system. Fifteen percent ethanolic extracts of mature leaves, young leaves and flowers of S. siamea were analyzed and compared for barakol content using the validated TLC-densitometric method. Both the validation and analysis of barakol by TLC-densitometry were carried out at the absorbance mode of 366 nm. RESULTS: Barakol was extracted as pure lemon-yellow crystals from young S. siamea leaves with 0.1% yield. Linearity was found over the range of 200-900 ng/spot (r(2) = 0.997). The developed method gave high precision (%RSD < 0.50) and accuracy (average 101.12%). The limit of detection and limit of quantitation were 8 and 50 ng, respectively. Barakol content in young leaves, mature leaves and young flowers were 1.67, 0.78 and 1.43% dry weight, respectively. R(f) value of the barakol in young leaves, young flowers and authentic sample was the same: 0.45 +/- 0.03. CONCLUSION: The TLC-densitometric method was simple, precise and convenient; hence it is an effective procedure for the simultaneous determination of barakol in plant extracts.

Barakol contents in fresh and cooked Senna siamea leaves.

Senna siamea (Lam.) Irwin and Barneby is a medicinal plant popularly used in Thailand. Young leaves and/or young flowers of this plant have been consumed by Thai people as a Khi Lek curry for a long time. The fresh young leaves and flowers are boiled with water 2-3 times to get rid of the bitterness and the boiled mush is used for curry cooking. Barakol, a major constituent of Senna siamea leaves was analyzed for its content in the fresh young leaves, the boiled leaves and the boiled filtrates by a high-performance thin-layer chromatographic method. Fresh young leaves of S. siamea contained 0.4035% w/w barakol. The amount of barakol in the first and second boiled filtrates were 0.2052 and 0.1079% fresh weight, while the first and second boiled leaves samples were 0.1408 and 0.0414% fresh weight, respectively. The results show the process of preparation of Khi Lek curry by boiling S. siamea young leaves twice with water reduced barakol content up to 90% and the content of barakol in boiled leaves used for curry has much less tendency to cause liver toxicity. This may explain the reason why Thai Khi Lek curry has not caused hepatotoxicity, unlike S. siamea leaves consumed as a powdered capsule.