**6. References**


[5] Shimanker K, Saxena B N & Fotherby K A (1978) Radioimmunoassay for serum medroxyprogesterone acetate. *J Steroid Biochem* 9(4), 359-63.

228 Chromatography – The Most Versatile Method of Chemical Analysis

P-DNPH: Prednisolone-2,4-Dinitrophenylhydrazone

Thailand, and the Hitachi Scholarship Foundation, Tokyo, Japan.

doses of Depoprovera. *Contraception* 22(5), 527-35.

cohort of adolescent girls. *J Adolescent Health* 35, 434-41.

MPA-DNPH: Medroxyprogesterone acetate-2,4-Dinitrophenylhydrazone

RP-HPLC-AD: Reversed phase high-performance liquid chromatography

*Department of Chemistry, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand* 

*Academic and Research Services Unit, Faculty of Pharmaceutical Sciences, Khon Kaen University,* 

The authors gratefully acknowledge the financial support from the National Research University (NRU) project, Khon Kaen University and the Center for Innovation in Chemistry (PERCH-CIC), Commission on Higher Education, Ministry of Education,

[1] Liskin L & Quillin W (1983) Long-acting progestins-promise and prospects. *Population* 

[2] Fotherby K, Koetsawang S & Mathrutham M (1980) Pharmacokinetic study of different

[3] Cromer B A, Stager M, Bonny A, Lazebnik R, Rome E, Ziegler J & Debanne S M (2004) Depot medroxyprogesterone acetate, oral contraceptives and bone mineral density in a

[4] Ohtsu T, Fujii H, Wakita H, Igarashi T, Itoh K & Imoto S (1998) Pharmacokinetic study of low-versus high-dose medroxyprogesterone acetate (MPA) in women. *Cancer* 

MPA: Medroxyprogesterone acetate

mg/mL: Milligram per milliliter

ng/mL: Nanogram per milliliter

 with amperometric detection RSD: Relative standard deviation

mM: Millimolar

ODS:Octadecylsilane

SD: Standard deviation SPE: Solid-phase extraction

M: Molar

**Author details** 

Saksit Chanthai *Corresponding Author* 

Thanee Tessiri

*Khon Kaen, Thailand* 

**6. References** 

*Reports* 2, K-18.

*Chemother Pharmacol* 42, 1-8.

**Acknowledgement** 

	- [19] Reich H, Grane K F & Sanfilippo S J (1953) The reaction of steroid ketones with 2,4 dinitrophenylhydrazine. *J Org Chem* 18, 818-32.

**Chapter 9** 

© 2012 HaiRu and Xiangyan, licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2012 HaiRu and Xiangyan, licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

)[18-20] , ammonium (NH4+)[12,21-23], and amino acids[24-27] from the external medium. However, for a long time, it has been unclear in what form nitrogen is translocated along the

distribution, and reproduction in any medium, provided the original work is properly cited.

**Chromatographic Analysis of Nitrogen** 

**Utilization and Transport in Arbuscular** 

The combination of chromatographic methods and mass spectrometry (MS) techniques is very useful in analysis of metabolites or components of complex biological samples. Chromatography has become the most important technique for separation and analyses, whereas MS is one of the most effective analytical methods used today for the determination of element concentrations, especially in the trace range; for the structural studies of organic and bioorganic compounds as well as isotopic analysis, due to its very high sensitivity, low detection limits and very small sample volumes needed[1-2]. Using chromatography and MS to separate and measure the concentration of amino acids has been well documented. Bengtsson[3] was the first to introduce a micro-method for the analysis of free amino acids in natural waters by gas chromatography (GC); the technique includes removal of interfering organic substances by chloroform extraction and purification of amino acids by cation exchange. Later, the procedure for analysis of amino acids from protein acid hydrolyzates as their *tert*-butyl dimethylsilyl derivatives by gas chromatography and mass spectrometry has been developed[4-9] and applied in biological matrix analysis[10-13]. Johansen et al.[12] and Rolin et al.[13] studied the nitrogen metabolism of external hyphae of the AM fungus using measurement of *tert*-butyldimethylsilyl(tBDMS)-derivatized amino acids levels by gas chromatography and mass spectrometry. Arbuscular mycorrhizal (AM) fungus is the oldest obligate symbiont,[14] benefiting the host plants by taking up N, P and other macronutrients, trace elements, and water from the soil. Among many components important for plant nutrition, nitrogen is often the most limiting but the AM fungi can improve the nitrogen (N) levels of their hosts.[15-17] The extraradical hyphae of the fungi effectively acquire nitrate

**Mycorrhizal Fungal Symbiosis** 

Additional information is available at the end of the chapter

Jin HaiRu and Jiang Xiangyan

http://dx.doi.org/10.5772/48469

**1. Introduction** 

(NO3-

