Determination of Thyroids and Steroids in Hair by High Performance Liquid Chromatography Tandem Mass Spectrometry

doi:10.19894/j.issn.1000-0518.220025

Chinese Journal of Applied Chemistry ›› 2022, Vol. 39 ›› Issue (11): 1703-1715.DOI: 10.19894/j.issn.1000-0518.220025

• Full Papers • Previous Articles Next Articles

Determination of Thyroids and Steroids in Hair by High Performance Liquid Chromatography Tandem Mass Spectrometry

Lin YUAN¹, Yan WU¹, Liu-Xi CHU¹, Wei WANG¹, Min-Hui ZHU¹, He ZHANG¹, Jin YANG², Hui-Hua DENG¹()

^1.Key Laboratory of Child Development and Learning Science，School of Biological Science and Medical Engineering，Southeast University，Nanjing 210096，China
^2.School of Public Health，Southeast University，Nanjing 210096，China

Received:2022-01-26 Accepted:2022-04-14 Published:2022-11-01 Online:2022-11-09
Contact: Hui-Hua DENG
About author:dengrcls@seu.edu.cn
Supported by:
the National Natural Science Foundation of China(81761128004)

Abstract

Abstract:

A sensitive and reliable method based on high performance liquid chromatography-tandem mass spectrometry （LC-MS/MS） is developed for the simultaneous determination of thyroids and steroids in human hair， including thyroxine （T4）， 3，3′，5-triiodothyronine （T3）， 3，3'，5'-triiodothyronine （rT3）， 3，3'-diiodothyronine （3，3'-T2） L-thyronine （T0）， cortisol （F） and cortisone （E）. The seven endogenous compounds are extracted from 20 mg hair sample incubated in the mixture solution of methanol and ammonia （volume ratio is 95∶5） for 24 hours at 40 ℃， and then analyzed with an electrospray ionization source in multiple reaction monitoring mode and positive ion mode. Under the isocratic elution conditions with methanol/pure water （volume ratio is 80∶20， containing 0.2 mmol/L ammonium acetate） as mobile phase， seven analytes are separated and well resolved. The limits of detection for the seven hormones range from 0.3 to 1.9 pg/mg and the limits of quantification from 1.1 to 6.5 pg/mg. The intra- and inter-day coefficients of variation are less than 10% and the recoveries range from 89.4% to 109.1% for all seven compounds. The stability and carryover also meet the detection criteria. The population analysis on hair samples from 140 participants reveals that T0， F and E could be quantified in all subjects， while T4， T3， rT3 and 3，3'-T2 could be detected in a small number of subjects， showing the contents around their limits of quantification.

Key words: Thyroids, Steroids, Liquid chromatography tandem mass spectrometry, Hair concentrations

CLC Number:

O657.6

Lin YUAN, Yan WU, Liu-Xi CHU, Wei WANG, Min-Hui ZHU, He ZHANG, Jin YANG, Hui-Hua DENG. Determination of Thyroids and Steroids in Hair by High Performance Liquid Chromatography Tandem Mass Spectrometry[J]. Chinese Journal of Applied Chemistry, 2022, 39(11): 1703-1715.

Figures/Tables 10

References 22

1	SELLMEYER D E， GRUNFELD C. Endocrine and metabolic disturbances in human immunodeficiency virus infection and the acquired immune deficiency syndrome［J］. Endocrine Rev， 1996， 17（5）： 518-532.
2	EPELBAUM J， TERRIEN J. Mini-review： aging of the neuroendocrine system： insights from nonhuman primate models［J］. Progn Neuro-Psychopharmacol Biol Psychiatry， 2020， 100： 109854.
3	JONGEJAN R M S， KLEIN T， MEIMA M E， et al. A mass spectrometry-based panel of nine thyroid hormone metabolites in human serum［J］. Clin Chem， 2020， 66（4）： 556-566.
4	VAN DER SPEK A H， FLIERS E， BOELEN A. The classic pathways of thyroid hormone metabolism［J］. Mol Cell Endocrinol， 2017， 458： 29-38.
5	GAO W， PENZ M， WEKENBORG M， et al. Determination of thyroid hormones in human hair with online SPE LC-MS/MS： analytical protocol and application in study of burnout［J］. Psychoneuroendocrinology， 2019， 106： 129-137.
6	TANOUE R， KUME I， YAMAMOTO Y， et al. Determination of free thyroid hormones in animal serum/plasma using ultrafiltration in combination with ultra-fast liquid chromatography-tandem mass spectrometry［J］. J Chromatogr A， 2018， 1539： 30-40.
7	武萌，韩建华，蔡洁，等. 妊娠早期甲状腺激素及尿碘参考值范围［J］. 卫生研究， 2017， 46（2）： 287-290.
	WU M， HAN J H， CAI J， et al. Investigation on reference ranges of thyroid hormones and urine iodine in early pregnancy［J］. Wei Sheng Yan Jiu， 2017， 46（2）： 287-290.
8	RICHARDS K H， MONK R， RENKO K， et al. A combined LC-MS/MS and LC-MS multi-method for the quantification of iodothyronines in human blood serum［J］. Anal Bioanal Chem， 2019， 411（21）： 5605-5616.
9	FISCHER S， STRAHLER J， MARKERT C， et al. Effects of acute psychosocial stress on the hypothalamic-pituitary-thyroid （HPT） axis in healthy women［J］. Psychoneuroendocrinology， 2019， 110： 104438.
10	KÖHRLE J， RICHARDS K H. Mass spectrometry-based determination of thyroid hormones and their metabolites in endocrine diagnostics and biomedical research-implications for human serum diagnostics［J］. Exp Clin Endocrinol Diabetes， 2020， 128（6/7）： 358-374.
11	LANGERAK T， VAN DEN DRIES L W， WESTER V L， et al. The relation between long-term cortisol levels and the metabolic syndrome in HIV-infected patients［J］. Clin Endocrinol （Oxf）， 2015， 83（2）： 167-172.
12	PATTERSON S， MORAN P， EPEL E， et al. Cortisol patterns are associated with T cell activation in HIV［J］. PLoS One， 2013， 8（7）： e63429.
13	WEI J， SUN G， ZHAO L， et al. Hair thyroid hormones concentration in patients with depression changes with disease episodes in female Chinese［J］. Psychiatry Res， 2014， 220（1/2）： 251-263.
14	KUNISUE T， FISHER J W， FATUYI B， et al. A method for the analysis of six thyroid hormones in thyroid gland by liquid chromatography-tandem mass spectrometry［J］. J Chromatogr B， 2010， 878（21）： 1725-1730.
15	CARROZZA C， LAPOLLA R， GERVASONI J， et al. Assessment of salivary free cortisol levels by liquid chromatography with tandem mass spectrometry （LC-MS/MS） in patients treated with mitotane［J］. Hormones， 2012， 11（3）： 344-349.
16	GROVA N， WANG X， HARDY E M， et al. Ultra-performance liquid chromatography-tandem mass spectrometer method applied to the analysis of both thyroid and steroid hormones in human hair［J］. J Chromatogr A， 2020， 1612： 460648.
17	TRIPATHY S K， AGRAWALA R K， BALIARSINHA A K. Endocrine alterations in HIV-infected patients［J］. Indian J Endocrinol Metab， 2015， 19（1）： 143-147.
18	DE LUCA R， DAVIS P J， LIN H Y， et al. Thyroid hormones interaction with immune response， inflammation and non-thyroidal illness syndrome［J］. Front Cell Dev Biol， 2020， 8： 614030.
19	ISLAM F M， WU J， JANSSON J， et al. Relative risk of renal disease among people living with HIV： a systematic review and meta-analysis［J］. BMC Public Health， 2012， 12： 234.
20	HANSEN M， LUONG X， SEDLAK D L， et al. Quantification of 11 thyroid hormones and associated metabolites in blood using isotope-dilution liquid chromatography tandem mass spectrometry［J］. Anal Bioanal Chem， 2016， 408（20）： 5429-5442.
21	PRAGST F， BALIKOVA M A. State of the art in hair analysis for detection of drug and alcohol abuse［J］. Clin Chim Acta， 2006， 370（1/2）： 17-49.
22	CHEN Z， LI J， XU G， et al. Simultaneous measurements of cortisol and cortisone in urine and hair for the assessment of 11β-hydroxysteroid dehydrogenase activity among methadone maintenance treatment patients with LC-ESI-MS/MS［J］. J Chromatogr B， 2014， 969： 77-84.

化合物 Compound	母离子 Precursor ion/Da	子离子 Product ion/Da	去簇电压 Declustering potential DP/V	入口电压 Entrance potential EP/eV	碰撞室入口电压 Collision cell entrance potential CEP/V	碰撞能 Collision energy CE/eV	碰撞室出口电压 Collision cell exit potential CXP/V
T4	777.2	731.2	68.0	7.6	57.5	32.0	7.0
T4^*	777.2	633.4	68.0	7.6	57.5	36.0	7.5
T4?¹³C₆	783.2	737.2	40.0	8.4	43.5	36.2	9.5
T3	651.5	605.3	41.0	8.4	41.8	30.7	6.2
T3^*	651.5	507.6	41.0	8.4	41.8	30.8	6.5
T3?¹³C₆	657.2	611.2	39.5	8.7	41.0	32.8	9.1
rT3	651.2	605.2	39.0	7.9	32.1	31.4	8.4
rT3^*	651.2	507.6	39.0	7.9	32.1	32.6	7.0
rT3?¹³C₆	657.1	611.3	34.0	8.4	54.5	31.0	8.5
3，3′?T2	525.6	479.6	30.7	6.9	56.1	28.3	7.5
3，3′?T2^*	525.6	381.9	30.7	6.9	56.1	29.5	5.6
3，3′?T2?¹³C₆	531.6	485.6	30.2	7.5	40.7	28.4	18.3
T0	274.0	215.0	51.0	5.8	37.5	24.5	1.2
T0^*	274.0	212.1	51.0	5.8	37.5	26.4	1.3
F	362.9	121.0	38.6	7.1	28.3	32.8	2.2
F^*	362.9	327.1	38.6	7.1	28.3	22.4	4.3
F?D2	365.0	122.0	41.6	6.4	30.4	38.1	1.4
E	360.9	163.0	48.9	7.8	26.4	31.6	0.9
E^*	360.9	121.1	48.9	7.8	26.4	43.2	0.8
E?D8	368.2	169.0	48.0	5.8	24.4	32.0	3.5

化合物 Compound	母离子 Precursor ion/Da	子离子 Product ion/Da	去簇电压 Declustering potential DP/V	入口电压 Entrance potential EP/eV	碰撞室入口电压 Collision cell entrance potential CEP/V	碰撞能 Collision energy CE/eV	碰撞室出口电压 Collision cell exit potential CXP/V
T4	777.2	731.2	68.0	7.6	57.5	32.0	7.0
T4^*	777.2	633.4	68.0	7.6	57.5	36.0	7.5
T4?¹³C₆	783.2	737.2	40.0	8.4	43.5	36.2	9.5
T3	651.5	605.3	41.0	8.4	41.8	30.7	6.2
T3^*	651.5	507.6	41.0	8.4	41.8	30.8	6.5
T3?¹³C₆	657.2	611.2	39.5	8.7	41.0	32.8	9.1
rT3	651.2	605.2	39.0	7.9	32.1	31.4	8.4
rT3^*	651.2	507.6	39.0	7.9	32.1	32.6	7.0
rT3?¹³C₆	657.1	611.3	34.0	8.4	54.5	31.0	8.5
3，3′?T2	525.6	479.6	30.7	6.9	56.1	28.3	7.5
3，3′?T2^*	525.6	381.9	30.7	6.9	56.1	29.5	5.6
3，3′?T2?¹³C₆	531.6	485.6	30.2	7.5	40.7	28.4	18.3
T0	274.0	215.0	51.0	5.8	37.5	24.5	1.2
T0^*	274.0	212.1	51.0	5.8	37.5	26.4	1.3
F	362.9	121.0	38.6	7.1	28.3	32.8	2.2
F^*	362.9	327.1	38.6	7.1	28.3	22.4	4.3
F?D2	365.0	122.0	41.6	6.4	30.4	38.1	1.4
E	360.9	163.0	48.9	7.8	26.4	31.6	0.9
E^*	360.9	121.1	48.9	7.8	26.4	43.2	0.8
E?D8	368.2	169.0	48.0	5.8	24.4	32.0	3.5

	平均值 Mean/%	标准差 SD/%
外标选择性 Selectivity for ESs
T4	4.6	0.4
rT3	7.5	1.2
T3	4.2	0.1
3，3'?T2	5.3	0.7
T0	7.2	1.2
F	8.8	1.3
E	5.7	1.1
内标选择性 Selectivity for ISs
T4?¹³C₆	0.5	0.3
rT3?¹³C₆	1.2	0.3
T3?¹³C₆	0.3	0.1
T2?¹³C₆	0.5	0.2
F?D2	1.6	0.5
E?D8	3.1	0.6

	平均值 Mean/%	标准差 SD/%
外标选择性 Selectivity for ESs
T4	4.6	0.4
rT3	7.5	1.2
T3	4.2	0.1
3，3'?T2	5.3	0.7
T0	7.2	1.2
F	8.8	1.3
E	5.7	1.1
内标选择性 Selectivity for ISs
T4?¹³C₆	0.5	0.3
rT3?¹³C₆	1.2	0.3
T3?¹³C₆	0.3	0.1
T2?¹³C₆	0.5	0.2
F?D2	1.6	0.5
E?D8	3.1	0.6

化合物 Compound	标准曲线 Standard curve	相关系数的平方（R²）	线性范围 Linear range/（pg·mg^-1）	检测限 LOD/（pg·mg^-1）	定量限 LOQ/（pg·mg^-1）
T4	y = 0.0016x-0.0035	0.996 7	6.5 ~ 1 000	1.9	6.5
rT3	y = 0.0063x-0.0111	0.999 7	3.9 ~ 1 000	1.2	3.9
T3	y = 0.0047x-0.0078	0.999 8	4.3 ~ 1 000	1.3	4.3
3，3'?T2	y = 0.0020x-0.0036	0.998 5	5.4 ~ 1 000	1.6	5.4
T0	y = 0.0021x-0.0037	0.997 2	3.9 ~ 1 000	1.2	3.9
F	y = 0.0167x-0.0098	0.996 2	1.8 ~ 1 000	0.6	1.8
E	y = 0.0623x-0.0373	0.999 2	1.1 ~ 1 000	0.3	1.1

Determination of Thyroids and Steroids in Hair by High Performance Liquid Chromatography Tandem Mass Spectrometry

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 22

Related Articles 2

Recommended Articles

Metrics

Comments

化合物 Compound	加标浓度 Spiked concentrations/（pg·mg^-1）	日内变异（n=5） Intraday CV/%	日间变异（n=5） Interday CV/%	回收率（M±SD） Recovery/%
T4	10	2.5	4.0	105.9±2.7
	20	7.0	5.0	102.0±7.2
	500	5.3	5.7	91.9±4.9
	800	2.9	3.3	94.7±2.8
rT3	8	3.9	3.2	98.9±3.8
	20	0.7	2.9	96.9±0.7
	500	3.0	6.9	92.1±2.8
	800	3.3	5.9	93.7±3.1
T3	10	2.4	2.5	105.8±2.5
	20	2.7	2.9	99.4±2.6
	500	4.1	5.3	93.3±3.8
	800	1.2	3.1	92.4±1.1
3，3'?T2	12	1.0	1.7	109.1±1.1
	20	7.0	5.7	98.9±6.9
	500	5.6	6.3	89.4±5.0
	800	4.7	5.2	92.1±4.4
T0	8	5.4	4.4	101.9±5.5
	20	2.3	3.2	99.1±2.3
	500	5.9	8.0	91.2±5.4
	800	4.6	4.4	94.4±4.3
F	2	7.1	7.2	103.2±7.3
	5	4.7	2.7	104.3±4.9
	500	8.3	6.1	96.0±8.0
	800	2.9	3.8	91.5±2.7
E	2	5.4	6.2	108.8±5.9
	5	4.9	2.9	101.2±5.0
	500	3.5	6.5	97.4±3.4
	800	3.9	3.1	96.4±3.7

化合物 Compound	外标残留效应 Carryoverfor ESs/%	内标残留效应 Carryoverfor ISs/%
T4	4.6	0.1
rT3	2.6	0.2
T3	1.4	0.1
3，3'?T2	1.7	0.1
T0	12.8	0.1
F	11.0	1.2
E	9.0	2.3

	加标浓度 Spiked concentrations/（pg·mg^-1）	平均值 Mean/（pg·mg^-1）	偏差 DEV/%	变异 CV/%
台式稳定性 Bench?top stability
T4	20	20.0	-0.2	0.5
T4	800	801.6	0.2	1.9
rT3	20	19.0	-5.0	4.9
rT3	800	790.1	-1.2	4.6
T3	20	20.4	2.1	5.5
T3	800	797.8	-0.3	8.2
3，3'?T2	20	20.4	2.0	6.3
3，3'?T2	800	739.8	-7.5	1.4
T0	20	19.5	1.1	4.3
T0	800	831.4	14.9	1.8
F	5	4.8	-4.5	12.5
F	800	756.5	-5.4	3.6
E	5	4.4	-12.3	1.5
E	800	728.4	-8.9	14.2
自动进样器稳定性 Auto?sampler stability
T4	20	18.3	-8.6	8.2
T4	800	788.0	-1.5	3.3
rT3	20	20.5	2.7	6.5
rT3	800	757.9	-5.3	5.2
T3	20	21.2	6.1	7.1
T3	800	747.2	-6.6	11.0
3，3'?T2	20	20.7	3.5	4.4
3，3'?T2	800	756.4	-3.6	3.1
T0	20	19.5	-2.7	3.9
T0	800	831.4	-9.9	4.0
F	5	4.9	-1.9	10.6
F	800	764.5	-4.4	2.6
E	5	4.5	-10.9	7.5
E	800	721.1	-9.9	4.3
储备液稳定性 Stock solutions stability
T4	20	20.3	1.4	1.2
T4	800	775.2	-3.1	0.8
rT3	20	19.5	-2.5	4.9
rT3	800	772.4	-3.5	1.8
T3	20	21.4	7.2	2.5
T3	800	809.3	1.2	0.6
3，3'?T2	20	20.9	4.5	7.0
3，3'?T2	800	761.1	-4.9	2.0
T0	20	18.9	-5.6	1.8
T0	800	803.1	-14.7	3.3
Continued on next page
F	5	4.8	-3.3	10.9
F	800	724.5	-9.4	4.1
E	5	5.3	6.9	4.1
E	800	739.4	-7.6	8.1
长期稳定性 Long?term stability
T4	20	18.9	-5.4	5.4
T4	800	751.8	-6.0	1.2
rT3	20	18.9	-5.3	6.1
rT3	800	791.7	-1.0	3.4
T3	20	20.2	0.8	8.1
T3	800	694.1	-13.2	1.2
3，3'?T2	20	20.0	2.6	3.0
3，3'?T2	800	607.7	-24.0	1.8
T0	20	18.9	-5.7	2.4
T0	800	897.4	12.2	7.3
F	5	4.8	-4.7	3.9
F	800	770.1	-3.7	4.2
E	5	5.3	5.4	5.1
E	800	757.5	-5.3	5.8
高温稳定性 High?temperature stability
T4	20	20.9	4.7	8.0
T4	800	800.7	0.1	2.5
rT3	20	19.8	-0.9	7.9
rT3	800	787.3	-1.6	3.3
T3	20	20.0	5.3	2.0
T3	800	815.3	1.9	9.2
3，3'?T2	20	20.7	3.5	8.3
3，3'?T2	800	779.6	-2.6	3.2
T0	20	19.4	-3.0	1.8
T0	800	789.9	-1.3	18.4
F	5	4.3	-13.3	2.7
F	800	766.0	-4.3	2.8
E	5	5.3	5.2	5.2
E	800	709.7	-11.3	8.1

[1]	WANG Wei, WU Yan, CHU Liu-Xi, YUAN Lin, ZHU Min-Hui, YANG Jin, DENG Hui-Hua. Detection of Endocannabinoids and Glucocorticoids in Hair of People Living with HIV by High Performance Liquid Chromatography Tandem Mass Spectrometry [J]. Chinese Journal of Applied Chemistry, 2021, 38(11): 1521-1530.
[2]	ZHANG Daihui1,2*, LU Lijun1, ZHANG Xun1, MU Jun1, ZHOU Xiao1, HU Tingting1, HAN Dachuan1. Simultaneous Determination of Ten Kinds of Perfluorinated Compounds in Lubricant Samples by High-performance Liquid Chromatography-Tandem Mass Spectrometry [J]. Chinese Journal of Applied Chemistry, 2012, 29(10): 1163-1170.