Method for synchronously determining angiotensin I and aldosterone in sample

A technology for angiotensin and aldosterone, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of large fluctuation range, poor specificity, affecting the sensitivity and specificity of clinical diagnosis, etc., and achieve high sensitivity, high specificity, high The effect of accuracy

Pending Publication Date: 2020-12-01
ZHONGSHAN HOSPITAL FUDAN UNIV +2
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AI Technical Summary

Problems solved by technology

Immunization method can realize automatic operation, fast detection and low cost, but only one target hormone can be detected at a time, and the specificity is poor, especially in female patients, cross-reaction is prone to occur
The level of endocrine hormones fluctuates widely in the pathological state of the body, and high / low values ​​are prone to appear. However, the accuracy and repeatability of the immunoassay method in such cases are insufficient, so that it cannot correctly reflect the patient's hormones in functional tests. The level fluctuates dynamically, thus affecting the sensitivity and specificity of clinical diagnosis
[0005] Existing mass spectrometry detection methods for PA screening are only suitable for the separate determination of a single target substance, and the pretreatment operations are complicated, or they are dedicated to the detection of other indicators
There is no report on simultaneous detection of Ang I and Ald in plasma

Method used

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  • Method for synchronously determining angiotensin I and aldosterone in sample
  • Method for synchronously determining angiotensin I and aldosterone in sample
  • Method for synchronously determining angiotensin I and aldosterone in sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1A

[0229] The detection of embodiment 1Ang I and Ald

[0230] (1) Reagent preparation

[0231] The internal standard adopted in the present embodiment is as follows: Aldosterone: Alderstone-D8, purchased from Cambridge Isotope Laboratories (Cambridge, MA, USA); Angiotensin I: Ang I-[13C6, 15N4], purchased from Cambridge Isotope Laboratories (Cambridge, MA , USA).

[0232] Preparation of standard song and internal standard stock solution : The standard substance of Ang I and the internal standard (Ang I-[13C6, 15N4]) standard substance were dissolved in deionized water respectively to obtain the standard substance stock solution and the internal standard stock solution of angiotensin I. Aldosterone standard and internal standard (Alderstone-D8) standard were dissolved in methanol to obtain aldosterone standard stock and internal standard stock solution. The stock solution is generally at the mg / mL level, such as 2mg / mL. Store the stock solution at -80°C after aliquoting.

[...

Embodiment 2

[0270] Embodiment 2: The method of the present invention is used for the diagnosis of hyperaldosteronism

[0271] (1) The patient is male. The patient complained of fatigue, unsteady walking, palpitations, headache, and cold sweat. The outpatient diagnosis was hypokalemia, hypertension, space-occupying lesions in the right kidney, and suspected primary aldosteronism. However, secondary hypertension due to primary aldosteronism was excluded as judged by confirmatory testing. The patient was eventually diagnosed with hypokalemia (renal potassium loss), bilateral renal cysts, and essential hypertension.

[0272] Take the patient's blood sample (plasma), and use the simultaneous measurement scheme described in the present invention for detection. The ratio ARR results of aldosterone and renin activity measured by sampling at different time points were all less than 30. An example of one of the measurement results is: aldosterone: 6.24ng / dL (aldosterone concentration is finally...

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PUM

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Abstract

The invention relates to a method for synchronously determining angiotensin I and aldosterone in a sample. Specifically, the invention provides the method for synchronously determining the contents ofangiotensin I (Ang I) and aldosterone (Ald) in a sample. The method comprises the following steps: (a) carrying out solid-phase extraction treatment on the sample; and (b) analyzing the sample treated in (a) through liquid chromatography tandem mass spectrometry, and determining the amount of Ang I and the amount of Ald therein, wherein the mass spectrum adopts a positive ion mode and negative ion mode switching mode for scanning. The invention also provides a system for synchronously determining angiotensin I and aldosterone content in the sample, and methods and systems for obtaining the aldosterone content and ratio (ARR)of the aldosterone to renin activity in the sample.

Description

technical field [0001] The present invention generally relates to the determination of angiotensin I and aldosterone in samples; in particular, it relates to the simultaneous determination of angiotensin I and aldosterone in samples by liquid chromatography-tandem mass spectrometry. Background technique [0002] The total number of hypertensive patients in China has exceeded 330 million, which can be divided into primary and secondary hypertension according to the etiology. Among them, primary aldosteronism (primary aldosteronism, PA) is the most common cause of secondary hypertension, accounting for about 10% of young hypertensive population. The clinical manifestation of PA is the clinical syndrome of hypertension and / or hypokalemia, which is a kind of "curable hypertension". It can reduce the incidence of cardiovascular events in patients and improve the prognosis of patients. [0003] Renin is a proteolytic enzyme released by the juxtaglomerular apparatus, which can ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/36G01N30/72G01N30/86G01N30/88
CPCG01N30/02G01N30/06G01N30/36G01N30/7266G01N30/8679G01N30/88G01N2030/062G01N2030/8813
Inventor 郭玮潘柏申李国庆王蓓丽陈方俊彭颖斐黄超刘丹王真昕胡玉懿程子韵张曦温冬
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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