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การประชุมวิชาการและนําเสนอผลงานระดับชาติ The 4th Annual Northeast Pharmacy Research Conference of 2012 11 – 12 กุมภาพันธ์ 2555 “Pharmacy Profession in Harmony” ณ คณะเภสัชศาสตร์ มหาวิทยาลัยขอนแก่น ______________________________________________________________________________________________  Determination of Tetracycline Using Green Reagent
Improvement of artemisinin production in tissue culture
of Artemisia annus L.
Introduction: Tetracycline is a broad-spectrum antibiotic
drug with activity against bacteria. It forms complex with Introduction: Artemisinin is an effective antimalarial
various metal ions such as calcium (II), magnesium (II), component isolated from Artemisia annua L., that recommended to artemisinin-based combination therapies copper (II), sodium (I) and iron (III). The complexation (ACTs) by the World Health Organization. Despite its low between metal ion and tetracycline could be used as basic content in part of leaf and shoot of artemisinin (0.01–1.1 % reaction for determination of tetracycline using DW) considerable efforts have been taken to develop more efficient ways of artemisinin production such as tissue culture spectrophotometric method. However, this method usually that can control environment condition and elicitation. uses chemicals as reagents which is caused toxicity in the Materials and methods: Callus of A.annua was grown in
environment. Materials and Method: Thus, this study used
Murashige and Skoog (MS) semisolid medium containing thidiazuron (TDZ), 1-naphthalene acetic acid (NAA) and mineral water which is come from natural source. This benzyladenine (BA) such as T0.1N1, T0.1N0.5, T0.5N0.5, reagent is contains with various minerals such as calcium T0.5N1, N0.5B1, N1B0.5 and N1B1. After 25 days of culture determine the artemisinin level by ELISA using anti- (II), magnesium (II), zinc (II), iron (II) and aluminium (III). artemisinin monoclonal antibody (MAb). Then cell suspension The optimum complexation ratio was mixed between was grown in MS with T0.5N0.5 for 25 days and added tetracycline and mineral water with a volume ratio of elicitor MJ, ABA dendrophthoe extract, cuscuta extract and its combination. After 6 days of elicitation determine 0.25:1, which was detected the absorption by UV-vis artemisinin level by ELISA technical. The results were spectrophometer on the range of 200-800 nm. Results:
calculated using One way ANOVA in 95% confidence intervals. Result: Callus cell that induce from A. annua
The optimum wavelength is 374 nm. The standard cultured in Murashige & Skoog (MS) semisolid media deviation of 7, 12, 25 μg ml-1 tetracycline were 0.00052, supplement with thidiazuron (TDZ) 0.5 mg/L, 1- 0.00092 and 0.00150, respectively. The percentage of naphthaleneacetic acid (NAA) 0.5 mg/L (T0.5 N0.5) produced maximum artemisinin production (455 ± 71.59 ug/g DW), eight time less than in vitro plantlets (3,537 ± 8.49 ug/g DW). respectively. Conclusion: The proposed method has been
After elicitation on 25 days old cell suspension in T0.5 N0.5, we found addition of 5.0 mg/l methyl jasmonate combination satisfactorily applied to determination of tetracycline from with 1.0 mg/l abscisic acid (in day 3) and 1 g/L dendrophthoe elicitor combination with 1.0 mg/l abscisic acid (in day 3) resulted in significant improvement of artemisinin production (12.29 ± 0.79 ug/g DW , 11.94 ± 0.61 ug/g DW, respectively) Keywords: Tetracycline, Mineral water, Green reagent,
comparison to control cultures (10.86 ± 0.75 ug/g DW, 2.53 ± 0.34 ug/g DW, respectively), which lower than callus cell that ____________________________________________________________ induced from A. annua. It may be due to cell suspension were second and third cycle, effect to artemisinin production. Conclusion: Callus cell in T0.5N0.5 was maximum growth
1Undergraduate student, Faculty of Pharmaceutical Sciences, and production and the result in combination of MJ with ABA and dendrophthoe elicitor with ABA is interesting to develop 2Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical for enhance artemisinin production in the future. *Corresponding author : E-mail: [email protected]
Keywords: artemisinin, cell suspension, dendrophthoe
____________________________________________________________ 1Student, Faculty of Pharmaceutical Science, Khon Kaen University
2Faculty of Pharmaceutical Science, Khon Kaen University
*Corresponding author: Tel (043) 272552 E-mail: [email protected]

Source: http://pharm.kku.ac.th/isan-journal/journal/volumn8-no1/4annual/078-KKU-KKU_Pages_290.pdf

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