Steroid Hormones LC-MS/MS Analysis Kit

STEROID HORMONES

Steroid Hormones in Serum LC-MS/MS Analysis Kit

Steroid hormones are defined as effective biological messengers which take part in crucial body processes even at low concentrations. Thus, reliable, and simultaneous analysis of a broad panel of steroid hormones plays vital role for the investigation of the hormone profile, which could be useful in detection endocrine disorders resulting from defects in steroid biosynthesis such as congenital adrenal hyperplasia (CAH). Due to the restrictions and shortcomings of immunoassay techniques for measurement of steroid hormones in body fluids, liquid chromatography tandem mass spectrometry (LC-MS/MS) is increasingly becoming the method for multi-class steroid hormone detection.

Highlights

Analysis of clinically relevant 16 steroid hormones in a single injection-no need for separate injections.

Total run time is 16.0 min.

One sample preparation for all steroid hormones. Easy liquid-liquid extraction step following by protein precipitation for the sample treatment-no need for SPE.

Covered corresponding isotope labeled internal standards of important steroid hormones-11 ISs.

Long life span of HPLC column.

Parameters

11-Deoxycorticosterone, 11-Deoxycortisol, 17-α-Hydroxypregnenolone, 17-α-Hydroxyprogesterone, Aldosterone, Androstene-3,17-dione, Androsterone, Corticosterone, Cortisol, DHEA, DHEAS, Dihydrotestosterone, Estradiol, Pregnenolone, Progesterone and Testosterone.

Matrix

Serum/Plasma

Sample Preparation

Step 1

Pipette 425 µl of serum/plasma sample, calibrators or controls into a glass centrifuge tube (tube-1) and add 25 µl of internal standard mixture, vortex for 5 sec.
Step 2

Add 450 µl of Reagent-1 into tube-1 and vortex for 15 seconds. Then, centrifuge at 4000 rpm for 5 min.
Step 3

Decant the supernatant into a new glass centrifuge tube (tube-2).
Step 4

Wash the precipitate in tube-1 with 1.5 mL Reagent-2 (two times!) and transfer the washing fluids into tube-2
Step 5

Agitate tube-2 for 3 min. and centrifuge at 4000 rpm for 5 min.
Step 6

Transfer 2.4 ml of the upper phase to a new glass tube (tube-3) and evaporate under nitrogen stream. (Upper phase can be evaporated at 7 psi during ~15 min using water bath at the temperature of 30 °C).
Step 7

Reconstitute tube-3 with 100 µl of Reagent-3 and transfer to a HPLC vial prior to injection.

Chromatogram

METHOD PERFORMANCE

Analyte	LOQ (µg/L) 6460 QQQ	LOQ (µg/L) Ultivo QQQ	Accuracy % 6460 QQQ	Repeatabilty % CV 6460 QQQ	Linearity R²6460 QQQ
11-Deoxycorticosterone	0,016	0,123	95.34	6.84	0.994
11-Deoxycortisol	0,015	0,049	102.23	4.45	0.998
17-Hydroxypregnenolone	0,145	1,251	100.44	9.24	0.997
17-Hydroxyprogesterone	0,028	0,078	97.01	4.80	0.997
4-Androstene 3,17-dione	0,023	0,193	109.93	1.99	0.998
Aldosterone	0,011	0,100	99.81	6.34	0.998
Androsterone	0,035	0,235	93.70	13	0.994
Corticosterone	0,054	0,424	98.42	4.09	0.998
Cortisol	0,102	0,694	89.74	1.69	0.999
DHEA	0,102	0,328	109.62	6.66	0.998
DHEAS	4,945	38,012	103.47	2.63	0.997
Dihydrotestosterone	0,012	0,263	101.98	14.36	0.997
Estradiol (Oestradiol)	0,015	0,227	109.37	6.32	0.998
Pregnenolone	0,033	0,370	77.49	10.46	0.994
Progesterone	0,033	0,079	108.37	4.45	0.994
Testosterone	0,008	0,027	96.75	1.80	0.997

Steroid Hormones in Serum LC-MS/MS Analysis Kit

Highlights

Parameters

Matrix

Sample Preparation

Chromatogram

METHOD PERFORMANCE

ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..

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