Establishment of traceable steroid hormone standard curve for dried blood spot samples

doi:10.3969/j.issn.1673-8640.2023.05.015

Abstract

Abstract:

Objective To establish a traceable standard curve for 19 steroid hormones in dried blood spot samples by liquid chromatography tandem mass spectrometry（LC-MS/MS）. Methods Ethylenediaminetetraacetic acid fresh anticoagulated whole blood was collected，blood cells were washed with 0.9 NaCl saline，and the washed blood cells and steroid hormone-free plasma were mixed in the ratio of 0.55∶0.45. Simulated whole blood samples was prepared. Different amounts of steroid hormones were added to simulated whole blood samples according to the prepared standard curve concentration in order to prepare different concentrations of steroid hormone standard curve dried blood spot samples. Steroid hormone concentrations in standard curve dried blood spot samples were determined by LC-MS/MS，and its linear range was analyzed. The performance of dried blood spot steroid hormone standard curve was verified. Results The linear ranges of dried blood spot steroid hormone standard curve for pregnenolone，progesterone，cortisone and dehydroepiandrosterone were 0.25-100 ng/mL. 11-Deoxycorticosterone，21-deoxycortisol，18-hydroxycortisol and 11-keto testosterone were in the linear range of 0.05-20 ng/mL. Corticosterone and 11-hydroxytestosterone were in the linear range of 0.025-10 ng/mL. 17-Hydroxyprogesterone，18-hydroxycorticosterone，11-deoxycortisol，androstenedione，11-hydroxyandrostenedione and testosterone were in the linear range of 0.1-40 ng/mL. Cortisol was in the linear range of 1.5-600 ng/mL，17-hydroxypregnenolone was in the linear range of 0.2-80 ng/mL，and dehydroepiandrosterone sulfate was in the linear range of 25-10 000 ng/mL with a correlation coefficient of squared（r²）≥0.997. Conclusions The established steroid hormone standard curves for dried blood spot samples have good linearities and can achieve the traceability of steroid hormone determination results in dried blood spots samples.

Key words: Steroid hormone, Standard curve, Dried blood spot, Liquid chromatography tandem mass spectrometry, Traceability

CLC Number:

R446.62

WU Jie, LI Zenghe, CHEN Baorong, KONG Xujing. Establishment of traceable steroid hormone standard curve for dried blood spot samples[J]. Laboratory Medicine, 2023, 38(5): 489-493.

Figures/Tables 3

References 9

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激素	线性范围/ （ng/mL）	斜率（a）	r²值
孕烯醇酮	0.250~100	0.030 5	0.998
17-羟孕烯醇酮	0.200~80	0.022 0	0.998
孕酮	0.250~100	0.222 1	0.998
17-羟孕酮	0.100~40	0.162 7	0.998
皮质酮	0.250~100	0.087 3	0.997
11-脱氧皮质酮	0.050~20	0.323 0	0.998
18-羟皮质酮	0.100~40	0.144 8	0.999
皮质醇	1.500~600	0.020 2	0.998
可的松	0.250~100	0.058 4	0.998
11-脱氧皮质醇	0.100~40	0.180 8	0.998
21-脱氧皮质醇	0.050~20	0.037 0	0.999
18-羟皮质醇	0.050~20	0.386 0	0.999
脱氢表雄酮	0.250~100	0.015 0	0.997
硫酸脱氢表雄酮	25.000~10 000	0.000 1	0.998
雄烯二酮	0.100~40	0.192 8	0.997
11-羟雄烯二酮	0.100~40	0.139 1	0.999
睾酮	0.100~40	0.270 3	0.998
11-羟睾酮	0.025~10	0.220 4	0.999
11-酮睾酮	0.050~20	0.753 8	0.998

激素	线性范围/ （ng/mL）	斜率（a）	r²值
孕烯醇酮	0.250~100	0.030 5	0.998
17-羟孕烯醇酮	0.200~80	0.022 0	0.998
孕酮	0.250~100	0.222 1	0.998
17-羟孕酮	0.100~40	0.162 7	0.998
皮质酮	0.250~100	0.087 3	0.997
11-脱氧皮质酮	0.050~20	0.323 0	0.998
18-羟皮质酮	0.100~40	0.144 8	0.999
皮质醇	1.500~600	0.020 2	0.998
可的松	0.250~100	0.058 4	0.998
11-脱氧皮质醇	0.100~40	0.180 8	0.998
21-脱氧皮质醇	0.050~20	0.037 0	0.999
18-羟皮质醇	0.050~20	0.386 0	0.999
脱氢表雄酮	0.250~100	0.015 0	0.997
硫酸脱氢表雄酮	25.000~10 000	0.000 1	0.998
雄烯二酮	0.100~40	0.192 8	0.997
11-羟雄烯二酮	0.100~40	0.139 1	0.999
睾酮	0.100~40	0.270 3	0.998
11-羟睾酮	0.025~10	0.220 4	0.999
11-酮睾酮	0.050~20	0.753 8	0.998

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