Development of highly sensitive quantification method for testosterone and dihydrotestosterone in human serum and prostate tissue by liquid chromatography–electrospray ionization tandem mass spectrometry

Yamashita, Kouwa; Miyashiro, Yoshimichi; Maekubo, Hitoe; Okuyama, Mitsunobu; Honma, Sato; Takahashi, Madoka; Numazawa, Mitsuteru

doi:10.1016/j.steroids.2009.06.007

Cited by 94 publications

(70 citation statements)

References 34 publications

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“…The E2 obtained was assayed by LC-MS/MS after derivatization, as described previously (Arai et al 2010). The androgen fraction was derivatized to the picolinoyl ester and assayed by LC-MS/MS (Yamashita et al 2009). The lowest analytical limits for DHEA, androstenedione, T, E2, and P4 were 10, 10, 2, 0.1, and 10 pg/assay, respectively.…”

Section: Methodsmentioning

confidence: 99%

Dioxin levels in the breast milk and estradiol and androgen levels in the saliva of Vietnamese primiparae

Nhu

Kido

Hưng

et al. 2011

Toxicological & Environmental Chemistry

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Human exposure to polychlorinated dibenzodioxins (PCDD) and dibenzofurans (PCDF), especially 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), was investigated in Vietnam since initial severe adverse health effects were reported in the late 1970s. The purpose of this study was to determine the effects of dioxin exposure on steroid hormones of primiparae in an Agent Orange/dioxin hot-spot and a non-exposed area in Vietnam. Sixteen primiparae (8 at each site), all of whom were aged between 20 and 30 years with infants aged between 4 and 16 weeks, agreed to participate in this study. The mean dioxin levels in breast milk of primiparae from the hot-spot area, in terms of PCDD, PCDF, and PCDD þ PCDF toxic equivalents (TEQ), were significantly higher than those for the non-exposed area. PCDD TEQ, PCDF TEQ, and PCDD þ PCDF TEQ levels showed a significant correlation with dehydroepiandrosterone (DHEA), androstenedione (A-dione), and estradiol (E2) in the saliva of primiparae in a combination of hot-spot and non-exposed areas in Vietnam. The dose-response curve between salivary E2 or A-dione levels and dioxin levels was U-shaped in humans. This study provides an overview of studies regarding dioxin hot-spots and effects on human health and steroid hormone levels in particular, with a focus on the toxicity attributed to dioxins and furans. Furthermore, causal evidence regarding the effects of dioxins on endocrine disruption in humans is provided.

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Section: Methodsmentioning

confidence: 99%

Dioxin levels in the breast milk and estradiol and androgen levels in the saliva of Vietnamese primiparae

Nhu

Kido

Hưng

et al. 2011

Toxicological & Environmental Chemistry

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“…It has generally been proposed that the presence of interference peaks co-existing in biological matrixes and co-eluting with the analyte of interest caused ionization suppression, reducing the detection sensitivity (12,13). For the measurement of steroids in organs, LC-MS analysis has several advantages over immunoassays, including a better specificity and the ability to quantify numerous steroids in a single run (14,15). Especially, liquid chromatographyelectrospray ionization time-of-flight mass spectrometry (LC-TOFMS) made possible to perform the accurate mass determination of components containing matrixes.…”

mentioning

confidence: 99%

Accurate determination of tissue steroid hormones, precursors and conjugates in adult male rat

Maeda

Tanaka

Suzuki

et al. 2012

The Journal of Biochemistry

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The actual levels of steroid hormones in organs are vital for endocrine, reproductive and neuronal health and disorders. We developed an accurate method to determine the levels of steroid hormones and steroid conjugates in various organs by an efficient preparation using a solid-phase-extraction cartridge. Each steroid was identified by the precursor ion spectra using liquid chromatographyelectrospray ionization timeof-flight mass spectrometry, and the respective steroids were quantitatively analysed in the selected reaction monitoring mode by liquid chromatograph-mass spectrometry/mass spectrometry (LC-MS/MS). The data showed that significant levels of testosterone, corticosterone and precursors of both hormones were detected in all organs except liver. The glucuronide conjugates of steroid hormones and the precursors were detected in all organs except liver, but sulfate conjugates of these steroids were observed only in the target organs of the hormones and kidney. Interestingly, these steroids and the conjugates were not observed in the liver except pregnenolone. In conclusion, an accurate determination of tissue steroids was developed using LC-MS analysis. Biosynthesis of steroid hormones from the precursors was estimated even in the target organs, and the delivery of these steroid conjugates was also suggested via the circulation without any significant hepatic participation.

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“…The reagent solution was used after 3-5 min [5]. Serum prostate-specific antigen (PSA) levels were measured using a DPC Imrise third generation PSA assay kit.…”

Section: Chemicals and Materialsmentioning

confidence: 99%

“…Serum hormone levels were determined by LC-MS/MS, as described by Yamashita et al [5]. T and DHT levels in prostate tissue samples were measured using an API-5000 triple-stage quadrupole mass spectrometer (Applied Biosystems, Foster City, CA, USA) connected to a Shimadzu LC-20 AD pump and Shimadzu SIL HTC autosampler (Shimadzu, Kyoto, Japan) and an electrospray ionization (ESI) ion-source device.…”

Section: Lc-ms/msmentioning

confidence: 99%

“…Advancements in liquid-chromatography tandem mass spectrometry (LC-MS/MS) methods mean that T and DHT levels can be measured in small tissue samples with high sensitivity and reliability [3][4][5][6]. LC-MS/MS can be used to measure androgen concentrations in tissue samples as small as those obtained by a single needle biopsy (approximately 3 mg), and the latest LC-MS/MS technique is more than 10 times as sensitive as the RIAs used in the past, especially in the lower concentration range [7].…”

Section: Introductionmentioning

confidence: 99%

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2128 High Testosterone Levels in Prostate Tissue Obtained by Needle Biopsy Correlate With Poor Prognosis Factors in Prostate Cancer Patients

Miyoshi¹,

Umemoto²,

Uemura³

et al. 2013

Journal of Urology

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Background: There is currently no consensus on the correlations between androgen concentrations in prostate tissue and blood and stage and pathological grade of prostate cancer. In this study, we used a newly-developed ultra-sensitive liquid-chromatography tandem mass spectrometry method to measure testosterone (T) and dihydrotestosterone (DHT) concentrations in blood and needle biopsy prostate specimens from patients with prostate cancer. Methods:We analyzed androgen levels in 196 men diagnosed with prostate cancer. All patients had undergone systematic needle biopsy, and an additional needle biopsy from the peripheral zone was conducted for the simultaneous determination of T and DHT. We analyzed the relationships between T and DHT levels in tissue and blood and Gleason score, clinical stage, and percentage of positive biopsy cores, using multivariate analysis. Results: The median T and DHT levels in blood were 3551.0 pg/mL and 330.5 pg/mL, respectively. There was a strong correlation between serum T and DHT. The median T and DHT levels in prostate tissue were 0.5667 pg/mg and 7.0625 pg/mg, respectively. In multivariate analysis, serum prostate-specific antigen and tissue T levels were significantly associated with poor prognosis; high T levels in prostate tissue were significantly related to high Gleason score (p = 0.041), advanced clinical stage (p = 0.002), and a high percentage of positive biopsy cores (p = 0.001). Conclusions:The results of this study indicate that high T levels in prostate tissue are related to high Gleason score, advanced clinical stage, and a high percentage of positive biopsy cores in patients with prostate cancer. T level in needle biopsy specimens may therefore be a useful prognostic factor in prostate cancer patients.

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Development of highly sensitive quantification method for testosterone and dihydrotestosterone in human serum and prostate tissue by liquid chromatography–electrospray ionization tandem mass spectrometry

Cited by 94 publications

References 34 publications

Dioxin levels in the breast milk and estradiol and androgen levels in the saliva of Vietnamese primiparae

Dioxin levels in the breast milk and estradiol and androgen levels in the saliva of Vietnamese primiparae

Accurate determination of tissue steroid hormones, precursors and conjugates in adult male rat

2128 High Testosterone Levels in Prostate Tissue Obtained by Needle Biopsy Correlate With Poor Prognosis Factors in Prostate Cancer Patients

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