การประชุมวิชาการและนําเสนอผลงานระดับชาติ The 4th Annual Northeast Pharmacy Research Conference of 2012 11 – 12 กุมภาพันธ์ 2555
“Pharmacy Profession in Harmony” ณ คณะเภสัชศาสตร์ มหาวิทยาลัยขอนแก่น
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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
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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
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1Student, Faculty of Pharmaceutical Science, Khon Kaen University 2Faculty of Pharmaceutical Science, Khon Kaen University *Corresponding author: Tel (043) 272552 E-mail: [email protected]
Toxicology and Industrial Health25(9-10) 711–714ª The Author(s) 2009Reprints and permission: http://www. sagepub.co.uk/journalsPermission.navDOI: 10.1177/0748233709348281tih.sagepub.comWilliam J Rea1, Yaqin Pan1 and Bertie Griffiths2AbstractTwenty-eight incapacitated individuals (average 43 years old, 7 males, 21 females, range 12-70) exposed tomolds and mycotoxins were studied and trea
Consultation response ‘Support for All: The Families and Relationships Green Paper’ Children England is the leading membership organisation for the children, young people and families’ voluntary sector. With member organisations working in all parts of the country ranging from small local groups to the largest household names in children’s charities, Children England is in a
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