Kit for detecting drug in sample and detection method thereof
The present application provides a kit for therapeutic drug monitoring, and a method and system thereof. A magnetic bead extraction method combined with LC-MS/MS detection is applied to drug concentration monitoring in human blood. A mixed magnetic bead solution containing a phospholipid depletion magnetic bead and a to-be-detected drug-adsorption magnetic bead is used for adsorption and extraction pretreatment of a drug to be detected in a sample. In conjunction with automated treatment equipment, the pretreatment efficiency is high such that the content of phospholipids in a supernatant is low, thereby greatly reducing the matrix effect and the chromatographic column flushing pressure. The method for therapeutic drug monitoring provided by the present application uses a unified pretreatment flow to achieve simultaneous extraction of multiple drugs. Moreover, by means of an automated magnetic bead extraction method, the steps for sample pretreatment can be completed rapidly, and the results for therapeutic drug monitoring can be obtained rapidly, such that the overall detection time is shortened, thereby providing a reliable laboratory examination basis for achieving individualized therapy.
1 . A method for drug detection, comprising the following steps:
making a mixed magnetic bead solution in contact with a sample, wherein the mixed magnetic bead solution comprises a magnetic bead for removing phospholipids in the sample and a magnetic bead for adsorbing a drug to be detected in the sample, such that the phospholipids and the drug to be detected in the sample are simultaneously adsorbed; and
eluting, with an eluent, the drug to be detected from the magnetic bead for adsorbing the drug to be detected in the sample, and analyzing a content of the drug to be detected by liquid chromatography-tandem mass spectrometry,
wherein the drug to be detected comprises any one or more of a sedative-hypnotic drug, an antidepressant drug, an antipsychotic drug, an antiepileptic drug, an antibiotic drug, an antineoplastic drug, a cardiovascular drug, and a toxicology screening drug;
wherein the magnetic bead for removing the phospholipids in the sample comprises a ZrO 2 silica magnetic bead, and the magnetic bead for adsorbing the drug to be detected in the sample comprises an HLB magnetic extraction bead; and
wherein the mixed magnetic bead solution comprises an acid, and the acid comprises any one or more of formic acid, acetic acid, and citric acid.
2 . The method according to claim 1 , wherein the sedative-hypnotic drug comprises any one or more of alprazolam, clonazepam, midazolam, lorazepam, zopiclone, temazepam, bromazepam, nitrazepam, 6-hydroxybuspirone, buspirone, zaleplon, memantine, donepezil, tandospirone, diazepam, nordazepam, oxazepam, zolpidem, and estazolam;
the antidepressant drug comprises any one or more of sertraline, fluoxetine, norfluoxetine, escitalopram, fluvoxamine, paroxetine, venlafaxine, O-demethyl-venlafaxine, duloxetine, mirtazapine, trazodone, milnacipran, amitriptyline, nortriptyline, doxepin, vortioxetine, norclomipramine, clomipramine, agomelatine, bupropion, mianserin, nordoxepin, and hydroxybupropion;
the antipsychotic drug comprises any one or more of olanzapine, clozapine, paliperidone, risperidone, dehydro-aripiprazole, aripiprazole, amisulpride, quetiapine, chlorpromazine, ziprasidone, norclozapine, haloperidol, perphenazine, sulpirida, norquetiapine, fluphenazine, thioridazine, atomoxetine, lurasidone, blonanserin, maprotiline, methylphenidate, rivastigmine, perospirone, norolanzapin, carbamazepine-10,11-epoxide, norsertraline, norcitalopram, and normirtazapine;
the antiepileptic drug comprises any one or more of oxcarbazepine, lamotrigine, levetiracetam, 10-OH Car, carbamazepine, phenytoin sodium, topiramate, primidone, gabapentin, pregabalin, rufinamide, striripentol, perampanel, zonisamide, lacosamide, valproic acid, and phenobarbital;
the antibiotic drug comprises any one or more of moxifloxacin, vancomycin, tigecycline, norvancomycin, polymyxin, linezolid, ciprofloxacin, sulfamethoxazole, and levofloxacin;
the antineoplastic drug comprises any one or more of cyclophosphamide, ifosfamide, methotrexate, 5-fluorouracil, capecitabine, irinotecan, paclitaxel, docetaxel, afatinib, rivoceranib, icotinib, erlotinib, gefitinib, crizotinib, regorafenib, vemurafenib, imatinib, N-desmethylimatinib, alectinib, and osimertinib;
the cardiovascular drug comprises any one or more of metoprolol, bisoprolol, nifedipine, amlodipine, atorvastatin, ortho-hydroxyatorvastatin, rosuvastatin, losartan, losartan-metabolite, valsartan, irbesartan, telmisartan, clopidogrel-metabolite, salicylic acid, ticagrelor, and ticagrelor-metabolite M8; and
the toxicology screening drug comprises any one or more of rodenticides selected from the group consisting of brodifacoum, bromadiolone, diphacinone, chlorophacinone, warfarin, flocoumafen, coumatetralyl, fluoroacetic acid, difenacoum, pindone, difethialone, coumafuryl, coumachlor, and melitoxin, pesticides, psychotropic drugs selected from the group consisting of piroxicam, 4-acetamidophenol, ethenzamide, paracetamol, sulindac, dihydroergotamine, ketorolac, ketoprofen, isopropylantipyrine, difenidol, loxapine, penfluridol, trihexylphenedyl, naproxen, N,N-diethylnicotinamide, benzoylecgonine, buprenorphine, fentanyl, flunitrazepam, ropivacaine, pethidine, procaine, oxycodone, tramadol, normorphine, ethylmorphine, dextropropoxyphene, and lidocaine, and biotoxins selected from the group consisting of aconitine, solanine, colchicine, amygdalin, ouabain octahydrate, tetrodotoxin, muscarine, and aflatoxin.
3 . The method according to claim 1 , wherein the eluent comprises methanol with formic acid.
4 . The method according to claim 1 , further providing an equilibrium solution, a diluent, and liquid chromatography mobile phase additives, wherein both the equilibrium solution and the diluent are aqueous formic acid solutions; the liquid chromatography mobile phase additives comprise an additive for mobile phase A and an additive for mobile phase B; the mobile phase A is an aqueous solution containing a mobile phase additive; the mobile phase B is a methanol solution containing a mobile phase additive; and the mobile phase additive is one or a combination of formic acid and ammonium acetate.