Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia Areata
Catecholamines and steroids are well-known neurotransmitters and hormones that rapidly change the excitability of neurons. Alopecia areata is a disease for which the exact cause is unknown, but it is considered to be associated with stress, and so the simultaneous analysis of catecholamines and ster...
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MDPI AG
2021-05-01
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| Online Access: | https://www.mdpi.com/1420-3049/26/9/2734 |
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| author | Yu-Ra Lee Bark-Lynn Lew Woo-Young Sim Jongki Hong Bong-Chul Chung |
| author_facet | Yu-Ra Lee Bark-Lynn Lew Woo-Young Sim Jongki Hong Bong-Chul Chung |
| author_sort | Yu-Ra Lee |
| collection | DOAJ |
| description | Catecholamines and steroids are well-known neurotransmitters and hormones that rapidly change the excitability of neurons. Alopecia areata is a disease for which the exact cause is unknown, but it is considered to be associated with stress, and so the simultaneous analysis of catecholamines and steroids is required for the diagnosis of alopecia areata. Thus, we herein report the simultaneous analysis of catecholamines and steroids bearing different functional groups for the first time, during which it was necessary to carry out a serial hydrolysis procedure. Following hydrolysis of the urine samples to produce the free forms from the urinary conjugates, ethyl acetate extractions were carried out, and chemical derivatization was performed using dansyl chloride to increase the sensitivity of the liquid chromatography–tandem mass spectrometry method. The matrix effects and recoveries of this analytical method were validated, giving values of 85.4–122.9% and 88.8–123.0%, respectively. In addition, the method accuracy and precision were assessed, giving values of 0.4–21.5% and 2.0–21.6% for the intra-day and inter-day precisions, respectively. This validated method was then applied to identify differences between patients with and without alopecia areata, wherein the metanephrine content was found to be significantly higher in the alopecia areata patient group. This quantitative profiling method can also be applied to steroid-dependent diseases, as well as catecholamine-related diseases. |
| first_indexed | 2024-03-10T11:39:50Z |
| format | Article |
| id | doaj.art-60a84a29058843df8ad0fd129765c7f4 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T11:39:50Z |
| publishDate | 2021-05-01 |
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| spelling | doaj.art-60a84a29058843df8ad0fd129765c7f42023-11-21T18:34:51ZengMDPI AGMolecules1420-30492021-05-01269273410.3390/molecules26092734Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia AreataYu-Ra Lee0Bark-Lynn Lew1Woo-Young Sim2Jongki Hong3Bong-Chul Chung4Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 02792, KoreaDepartment of Dermatology, Kyung Hee University Hospital at Gangdong, Kyung Hee University, Seoul 05278, KoreaDepartment of Dermatology, Kyung Hee University Hospital at Gangdong, Kyung Hee University, Seoul 05278, KoreaKHU-KIST Department of Converging Science and Technology, Kyung Hee University, Seoul 02447, KoreaMolecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 02792, KoreaCatecholamines and steroids are well-known neurotransmitters and hormones that rapidly change the excitability of neurons. Alopecia areata is a disease for which the exact cause is unknown, but it is considered to be associated with stress, and so the simultaneous analysis of catecholamines and steroids is required for the diagnosis of alopecia areata. Thus, we herein report the simultaneous analysis of catecholamines and steroids bearing different functional groups for the first time, during which it was necessary to carry out a serial hydrolysis procedure. Following hydrolysis of the urine samples to produce the free forms from the urinary conjugates, ethyl acetate extractions were carried out, and chemical derivatization was performed using dansyl chloride to increase the sensitivity of the liquid chromatography–tandem mass spectrometry method. The matrix effects and recoveries of this analytical method were validated, giving values of 85.4–122.9% and 88.8–123.0%, respectively. In addition, the method accuracy and precision were assessed, giving values of 0.4–21.5% and 2.0–21.6% for the intra-day and inter-day precisions, respectively. This validated method was then applied to identify differences between patients with and without alopecia areata, wherein the metanephrine content was found to be significantly higher in the alopecia areata patient group. This quantitative profiling method can also be applied to steroid-dependent diseases, as well as catecholamine-related diseases.https://www.mdpi.com/1420-3049/26/9/2734alopecia areatacatecholaminemetanephrinesteroidliquid chromatography–tandem mass spectrometry |
| spellingShingle | Yu-Ra Lee Bark-Lynn Lew Woo-Young Sim Jongki Hong Bong-Chul Chung Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia Areata Molecules alopecia areata catecholamine metanephrine steroid liquid chromatography–tandem mass spectrometry |
| title | Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia Areata |
| title_full | Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia Areata |
| title_fullStr | Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia Areata |
| title_full_unstemmed | Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia Areata |
| title_short | Serial Hydrolysis for the Simultaneous Analysis of Catecholamines and Steroids in the Urine of Patients with Alopecia Areata |
| title_sort | serial hydrolysis for the simultaneous analysis of catecholamines and steroids in the urine of patients with alopecia areata |
| topic | alopecia areata catecholamine metanephrine steroid liquid chromatography–tandem mass spectrometry |
| url | https://www.mdpi.com/1420-3049/26/9/2734 |
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