2-Hydrazino-1-methylpyridine: a highly sensitive derivatization reagent for oxosteroids in liquid chromatography–electrospray ionization-mass spectrometry

Higashi, Tatsuya; Yamauchi, Akinori; Shimada, Kaoru

doi:10.1016/j.jchromb.2004.12.007

Cited by 92 publications

(80 citation statements)

References 12 publications

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“…However, it is also true that the introduction of charged moieties enhances the ionization efficiency of neutral steroids in ESI and atmospheric pressure chemical ionization processes. A permanently charged Nmethylpyridyl group was introduced into the hydroxyl group of cholesterol (23) and 5a-dihydrotestosterone (24,25), whereas a permanently charged quaternary pyridinium moiety was introduced to the carbonyl group of oxosteroids with Girard P reagent (26-28) or 2-hydrazino-1-methylpyridine (29). Very recently, Yamashita et al (19) developed a new picolinoyl derivatization that is a simple and versatile method suitable for the sensitive and specific quantification of hydroxysteroids by positive ESI-LC-MS/ MS. Interestingly, because the picolinoyl group is not charged permanently, even in the case of estradiol with two hydroxyl groups, a single charged ion was observed in the positive ESI mass spectrum.…”

Section: Discussionmentioning

confidence: 99%

Highly sensitive quantification of 7α-hydroxy-4-cholesten-3-one in human serum by LC-ESI-MS/MS

Honda

Yamashita

Numazawa

et al. 2007

Journal of Lipid Research

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We describe a highly sensitive and specific method for the quantification of serum 7a-hydroxy-4-cholesten-3-one (C4), which has been used as a biomarker for bile acid biosynthesis. This method is based upon a stable isotope dilution technique by liquid chromatography-tandem mass spectrometry (LC-MS/MS). C4 was extracted from human serum (2-50 ml) by a salting-out procedure, derivatized into the picolinoyl ester (C4-7a-picolinate), and then purified using a disposable C 18 cartridge. The resulting picolinoyl ester derivative of C4 was quantified by LC-MS/MS using the electrospray ionization mode. The detection limit of the C4 picolinoyl ester was found to be 100 fg (signal-to-noise ratio 5 10), which was z1,000 times more sensitive than the detection limit of C4 with a conventional HPLC-ultraviolet method. The relative standard deviations between sample preparations and between measurements by our method were calculated to be 5.7% and 3.9%, respectively, by one-way layout analysis. The recovery experiments were performed using serum spiked with 20.0-60.0 ng/ml C4 and were validated by a polynomial equation. The results showed that the estimated concentration with 95% confidence limit was 23.1 6 2.8 ng/ml, which coincided completely with the observed X 0 6 SD 5 23.3 6 1.0 ng/ml with a mean recovery of 93.4%. This method provides highly reliable and reproducible results for the quantification of C4, especially in small volumes of blood samples.

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Section: Discussionmentioning

confidence: 99%

Highly sensitive quantification of 7α-hydroxy-4-cholesten-3-one in human serum by LC-ESI-MS/MS

Honda

Yamashita

Numazawa

et al. 2007

Journal of Lipid Research

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“…11,12) Therefore, we examined the product ions of the bisHP and bisNA derivatives during MS-MS, because they will be employed as the monitoring ions in future biological sample analyses with the SRM mode. The bisHP derivatives of AD and PROG provided a satisfactorily intense product ion at m/z 322 and 348, respectively.…”

Section: Advantage Of Hp-and Na-derivatives In Srm Modementioning

confidence: 99%

Alternative Procedure for Charged Derivatization to Enhance Detection Responses of Steroids in Electrospray Ionization-MS

Higashi

Nishio

Hayashi

et al. 2007

CHEMICAL & PHARMACEUTICAL BULLETIN

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A derivatization procedure has been examined to enhance the electrospray ionization (ESI)-MS detectabilities of steroids that charged derivatization is not suitable for. The derivatization procedure with 2-hydrazinopyridine or isonicotinoyl azide was very effective for the sensitive detection of di-oxosteroids or di-hydroxysteroids, respectively, and the detection limits of the resulting derivatives were as low as about 2 fmol. The derivatives also provided intense characteristic product ions in the MS-MS, which are expected to be usable for the selected reaction monitoring mode.

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“…[11][12][13][14][15][16][17][18] In our previous studies, we have demonstrated that the picolinyl derivatization of hydroxyl group in the steroid molecules proceeded smoothly and quantitatively, and the picolinyl derivative thus obtained by simple one-step procedure exhibited preferable mass spectral advantages in enhancing sensitivity to LC-ESI-MS/MS. [19][20][21][22] The wide application of picolinyl derivatization to steroid hydroxy group irrespective of positional and/or steric factors is another great advantage of this reagent 19) when compared to recently developed reagents.…”

Section: Notesmentioning

confidence: 99%

Preparation and Structural Elucidation of the Picolinyl Ester of Aldosterone for Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

Yamashita

Tadokoro

Takahashi

et al. 2008

CHEMICAL & PHARMACEUTICAL BULLETIN

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2-Hydrazino-1-methylpyridine: a highly sensitive derivatization reagent for oxosteroids in liquid chromatography–electrospray ionization-mass spectrometry

Cited by 92 publications

References 12 publications

Highly sensitive quantification of 7α-hydroxy-4-cholesten-3-one in human serum by LC-ESI-MS/MS

Highly sensitive quantification of 7α-hydroxy-4-cholesten-3-one in human serum by LC-ESI-MS/MS

Alternative Procedure for Charged Derivatization to Enhance Detection Responses of Steroids in Electrospray Ionization-MS

Preparation and Structural Elucidation of the Picolinyl Ester of Aldosterone for Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

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