Method and kit for detecting five immunosuppressants in dried blood spot

Abstract

The invention discloses a liquid chromatography-tandem mass spectrometry method and a kit for accurately determining the content of five immunosuppressants in a dry blood spot. The method comprises the following steps: 1) uniformly mixing an internal standard working solution with an organic solvent to obtain an extracting solution; 2) extracting the dry blood spot of the sample to be detected by using the extracting solution, centrifuging to obtain a supernatant I after the extraction is finished, blow-drying the supernatant I with nitrogen, re-dissolving to obtain a solution II, whirling, centrifuging, and detecting; and 3) detecting the solution II by adopting a liquid chromatography-tandem mass spectrometry method, wherein the internal standard working solution contains a cyclosporin A isotope internal standard substance, an everolimus isotope internal standard substance, a sirolimus isotope internal standard substance and a mycophenolic acid isotope mixed internal standard substance. According to the method, during detection, five immunosuppressants can be quantified at a time only by 6 microliters of a blood sample, the pretreatment method is simple and rapid, the operation time is saved, the immunosuppressive blood concentration can be effectively monitored according to the indexes, and more complete treatment medication information can be obtained in time.

Description

technical field [0001] The invention relates to a method for detecting five kinds of immunosuppressants in dried blood slices and a kit thereof. Background technique [0002] Immunosuppressants are a class of chemical or biological substances that reduce tissue damage by inhibiting cellular and humoral immune responses, and can inhibit abnormal immune responses in the body. They are mainly used in the treatment of organ transplantation anti-rejection and autoimmune diseases [1] . Since the development of immunosuppressants in the 1970s, the immunosuppressants currently being clinically used can basically be divided into five categories. The first category is cytokine inhibitors, such as cyclosporine A, tacrolimus, sirolimus, everolimus and so on. The second category is DNA synthesis inhibitors, such as azathioprine, mycophenolic acid, mizoribine and so on. The third category is glucocorticoids, such as prednisone and methylprednisone. The fourth category is anti-lymphocy...

AI Technical Summary

Problems solved by technology

[0004] The analytes detected by the kits of the existing LC-MS / MS method are not comprehensive, including only four immunosuppressants CsA, Tac, Sir, and Ever, and do not contain mycophenolic acid (MPA), which is commonly used in clinical practice. Inability to perform concurrent therapeutic drug monitoring of five immunosuppressants commonly involved

[0005] At the same time, it is worthy of our attention that during the period of immunosuppressant TDM, whole blood needs to be collected multiple times, which increases the physiological and economic burden of patients and their families.

Images