Determination of 9 Amino Acid Endogenous Substances in Morning Urine of Depression Patients by LC-MS/MS
- VernacularTitle:LC-MS/MS法测定抑郁症患者晨尿中9种氨基酸类内源性物质的浓度
- Author:
Hongyan SONG
1
;
Jingsheng XIA
1
;
Gangjian JI
1
;
Zhangtao REN
1
;
Xianglin ZHANG
2
;
Fang LIU
2
;
Ling CHEN
1
Author Information
1. Dept. of Pharmacy，Wuhan Third Hospital/Tongren Hospital Affiliated to Wuhan University，Wuhan 430060，China
2. Dept. of Pharmacy，China-Japan Friendship Hospital，Beijing 100029，China
- Publication Type:Journal Article
- Keywords:
Depression patients;
Urine concentration;
LC-MS/MS;
Amino acid substances
- From:
China Pharmacy
2020;31(1):91-98
- CountryChina
- Language:Chinese
-
Abstract:
ABSTRACT OBJECTIVE：To establish a method for the determination of 5-hydroxyindole acetic acid（5-HIAA），glutamine， hippurate，pimelate，proline，tryptophan，tyramine，tyrosine and valine in human urine. METHODS：Morning urine samples were collected from depression patients. The sample was extracted with acetonitrile after addition of internal standard cortisone. LC-MS/ MS method was adopted. The determination was performed on XTerra RP18 column with mobile phase consisted of 0.1％ acetic acid-water as mobile phase A and 0.1％ acetic acid-acetonitrile as mobile phase B（gradient elution）. The column temperature was set at 40 ℃，and flow rate was 0.45 mL/min. Electrospray ion source（ESI）was used for quantitative analysis by multiple reaction monitoring （MRM）. The precursor/product ion transitions （m/z） were monitored at m/z 192.2→146.1，m/z 147.2→130.0，m/z 180.1→105.1，m/z 161.1→125.2，m/z 116.1→70.2，m/z 205.2→188.2，m/z 138.2→121.1，m/z 182.0→123.0，m/z 118.2→72.1 and m/z 361.2→163.0（+ion-mode）for 5-HIAA，glutamine，hippurate，pimelate，proline，tryptophan，tyramine，tyrosine，valine and cortisone，respectively. RESULTS：The linear range of 5-HIAA，glutamine，pimelate，proline，tyramine and valine were 10.00-3 200 ng/mL（r＝0.993 8-0.998 9，n＝6）. The lower limit of quantification was 10 ng/mL；the linear range of hippurate，tryptophan and tyrosine were 1 600-51 200 ng/mL（r＝0.999 2-0.999 7，n＝6）. The lower limit of quantitation was 1 600 ng/mL. The results of accuracy tests were 86.29％-98.65％（n＝6）. RSDs of intra-day and inter-day precision tests were no more than 14.65％（n＝6）CV of matrix effect were 6.18％-14.37％（n＝6）. Extraction recovery rates were 86.21％-98.14％（n＝6）. RE of stability tests were no more than 14.71％（n＝3-6）. CONCLUSIONS：The method is sensitive，accurate and suitable for the determination of 9 substances in human.
