Bioanalytical Method Validation Research Articles

A Selective and Accurate LC-MS/MS Method for Simultaneous Quantification of Valsartan and Hydrochlorothiazide in Human Plasma

The fixed dose combination of valsartan (VAL) and hydrochlorothiazide (HCTZ) is the most commonly prescribed medicine for the effective treatment of hypertension. In this study, a simple sensitive and accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous quantitation of VAL and HCTZ in human plasma by using irbesartan (IRB) and hydroflumethiazide (HFMZ) as their specific internal standards (ISs). HLB cartridge-based solid-phase extraction was used for the extraction of analytes and ISs. The chromatographic separation was achieved on Lichrocart RP Select (125 × 4 mm), 5 nm with the mobile phase composition of acetonitrile: 10 mM ammonium acetate buffer: 95:05, v/v, at flow rate of 0.5 mL/min. The turbo ion electrospray ionization in negative mode was used as ion source for the sample ionization. The precursor to product ion transitions were 434.10 > 179.10 (VAL), 295.70 > 204.90 (HCTZ), 427.10 > 192.90 (IRB), and 329.90 > 302.40 (HFMZ) for detection and quantification of analytes and their ISs. The retention times of VAL and HCTZ were 1.90 min and 2.30 min, respectively. The range for the calibration curves of VAL and HCTZ were 50.2–6018.6 ng/mL and 1.25–507.63 ng/mL, respectively, with good linearity having correlation coefficient values of ≥0.995 for both VAL and HCTZ. All validation parameter results (selectivity, precision and accuracy, matrix effects and stabilities) were within the acceptable range as per USFDA guideline for bioanalytical method validation. The intra-day and inter-day accuracy data for VAL were within the range of 105.68–114.22% and 98.41–108.16%, respectively, whereas for HCTZ they were 87.01–101.18% and 95.16–99.37%, respectively. The ion suppression effects produced for VAL and ion enhancement effects produced for HCTZ were insignificant according to the proposed sample cleanup procedure. The developed LC-MS/MS method was successfully applied to bioequivalence study on healthy volunteers.

A Bioanalytical Method for Quantification of N-nitrosodimethylamine (NDMA) in Human Plasma and Urine with Different Meals and following Administration of Ranitidine

Control of N-nitrosoamine impurities is important for ensuring the safety of drug products. Findings of nitrosamine impurities in some drug products led FDA to develop new guidance providing recommendations for manufacturers towards prevention and detection of nitrosamine impurities in pharmaceutical products. One of these products, ranitidine, also had a published in vivo study, which has since been retracted by its authors, suggesting a potential for in vivo conversion of ranitidine to the probable human carcinogen, N-nitrosodimethylamine (NDMA). FDA subsequently initiated a randomized, double-blind, placebo-controlled, crossover clinical investigation to assess the potential for in vivo conversion of ranitidine to NDMA with different meals. A bioanalytical method toward characterization of NDMA formation was needed as previously published methods did not address potential NDMA formation after biofluid collection. Therefore, a bioanalytical method was developed and validated as per FDA's Bioanalytical Method Validation guidance. An appropriate surrogate matrix for calibration standards and quality control sample preparation for both liquid matrices (human plasma and urine) was optimized to minimize the artifacts of assay measurements and monitor basal NDMA levels. Interconversion potential of ranitidine to NDMA was monitored during method validation by incorporating the appropriate quality control samples. The validated methods for NDMA were linear from 15.6 pg/mL to 2000 pg/mL. Low sample volumes (2 mL for urine and 1 mL for plasma) made this method suitable for clinical study samples and helped to evaluate the influence of ranitidine administration and meal types on urinary excretion of NDMA in human subjects.

A Rapid Method for Determination of Serum Methotrexate Using Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry and Its Application in Therapeutic Drug Monitoring.

Objectives Methotrexate (MTX) has anticancer therapeutic potential with multiple doses-related adverse effects and toxicities. Immunoassays for therapeutic monitoring of serum MTX have their own limitations. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is considered as the reference method; however, commercially availability of them is limited. We aimed to adapt/develop an in-house LC-MS/MS method for therapeutic monitoring of serum MTX. Materials and Methods Serum protein precipitation was performed using acetonitrile-water containing 250 μM solution of aminoacetophenone as internal standard (IS). Chromatographic separation was achieved on a C18 column with mobile phase of 0.1% solution of formic acid (solvent A) and acetonitrile (solvent B) at a flow rate of 0.4 mL/min. MS was performed under positive ion mode with mass transition for MTX and IS as m/z 455.1→308.1 and 136.2→94.1, respectively. The method was validated by following Bioanalytical Method Validation Guidance for Industry, 2018 and applied on leukemia patients' samples on MTX therapy. Results The correlation coefficient of eight serially diluted calibration standards of 0.09 to 12.5 μM was >0.99 and had linearity with > 95% precision and accuracy at analytical quality control levels. The lower limit of MTX quantification achieved was 0.09 μM with good intensity and sharp peak as compared with blank sample. The total run time of the assay was 5 minutes. The serum MTX levels obtained by this method in leukemia patients exhibited clinical correlation and an excellent agreement with commercial immunoassay used in parallel. Conclusion We were able to develop a rapid, sensitive, and cost-effective LC-MS/MS method suitable for therapeutic drug monitoring of MTX in routine clinical diagnostic laboratories.

Bioanalytical Method Development and Validation for Determination of Rifampicin and Quercetin in Rat Plasma by UHPLC-MS/MS: Applications to Pharmacokinetic Study

UHPLC-MS/MS was utilised to quantitatively analyse Rifampicin and Quercetin-loaded Liquisolid compact in rat plasma. The UPLC Acquity C18 (1.7 μm, 2.1 × 15 mm) column and 0.5 ml/min were used to separate the analyte. In a low-pressure gradient mode, A: In water 0.1 % formic acid and B: In acetonitrile 0.1 % formic acid as the mobile phase. Sample pre-treatment was performed by protein precipitation technique with methanol and acetonitrile (1:1) from rat plasma. As an internal standard (IS), the analyte was found by tracking precursor-to-product ion transformations of 823 → 791.3 m/z for rifampicin, 303 → 257 m/z for quercetin, and 138 → 121 m/z for isoniazid in MRM mode. The proposed technique was validated for precisions (Intraday and Interday) accuracy, linearity, quantification at lower limits, and analyte recovery. The findings showed that the plasma samples inter and intra-day precision and consistency values were determined to be within the acceptable range. After orally administering Liquid solid compact and pure drug solution, which showed a substantial difference in the pace and extent of absorption, the method’s suitability for determining the pharmacokinetic profile of each drug was tested.

Development and Validation of a Novel Bioanalytical Method for Estimating Epigallocatechin 3 Gallate in Wistar Rat Plasma by RP-HPLC Employing Gradient Elution Techniques

Development and Validation of a Novel Bioanalytical Method for Estimating Epigallocatechin 3 Gallate in Wistar Rat Plasma by RP-HPLC Employing Gradient Elution Techniques

Bio-Analytical Method Development and Validation of Asciminib and its Application to Pharmacokinetic Studies in Rat Plasma by Using RP-HPLC

The aim of the proposed study was to validate a rapid, uncomplicated, accurate, robust and sensitive bio-analytical method for quantifying asciminib using ivosidenib as an internal standard in rat plasma. This work provides a summary of recent advances in bio-analytical HPLC methodologies employing a Luna phenyl hexyl column (150 × 4.6 mm, 3.5) with an organic mobile phase of potassium dihydrogen phosphate (adjusted to pH 3.0 with orthophosphoric acid) and acetonitrile in the ratio 40:60. With a correlation coefficient of 0.999, the asciminib calibration curve was linear over the range of 100-2000 ng/mL. Using liquid-liquid extraction, the percent of recovered ASB ranged between 97.8 and 99.9%. The plasma stability of the bench-top and auto sampler remained stable for 24 h in the auto sampler. The T1/2 is 5 h and Tmax is 3 h, as calculated from the pharmacokinetic parameters. The application indicates that all parameters of system suitability, specificity, linearity and accuracy are in good agreement with the prescribed limits; consequently, the current method is highly stable, rapid and capable of analyzing plasma samples. This method is ideally adapted for determining asciminib in pure or dose form due to its novel technology.

Bioanalytical method development and validation of folic acid from rat plasma using reverse phase high performance liquid chromatography

Bioanalytical method development and validation of folic acid from rat plasma using reverse phase high performance liquid chromatography

Bioanalytical method development and validation of a novel antiseizure agent Cenobamate using LC-MS/MS

Bioanalytical method development and validation of a novel antiseizure agent Cenobamate using LC-MS/MS

Bio-Analytical Method Development and Validation for Ramipril and Its Metabolite by Liquid Chromatography-Tandem Mass Spectrometry

Bio-Analytical Method Development and Validation for Ramipril and Its Metabolite by Liquid Chromatography-Tandem Mass Spectrometry

Bioanalytical method development and validation for the quantitation of larotrectinib in human plasma: Application to pharmacokinetics in healthy rabbits

Bioanalytical method development and validation for the quantitation of larotrectinib in human plasma: Application to pharmacokinetics in healthy rabbits

Development and Validation of Bioanalytical Method for the Quantification of Febuxostat in Human K2 EDTA Plasma by LC-MS/MS: Pharmacokinetic Studies in Wister Rats

A specific, linear and accurate LC-MS/MS method was developed for the determination of febuxostat in human K2 EDTA plasma and it was successfully applied for the pharmacokinetic study in wister rats. Chromatographic isolation of febuxostat and febuxostat D9 were attained on Purosphur C18, 100 × 4.6 mm, 5 μ column with 1.0 mL/min flowing rate. The technique was linear over the standard concentrations ranging from 24.995-7001.401 ng/mL and the regression coefficient was perceived to be ≥ 0.9997. All LLOQ samples % RSD of back computed concentrations varied from 1.00 to 4.01. The analysis of plasma from healthy rats was successfully conducted using the validated LC-MS/MS procedure to quantify the presence of febuxostat. From the pharmacokinetic studies, Tmax, Cmax, T1/2 and AUC0-∞ of the febuxostat tablets were 2.0 ± 0.03 h, 8.85 ± 1.87 ng/mL, 6.34 ± 0.53 h, 95.58 ± 6.37 ng h/mL, respectively.

UPLC–MS/MS-Based Targeted Proteomics Assay to Evaluate the Relationship between CYP3A2 Expression and Enzymatic Activity

Traditional analytical methods such as western blotting and reverse transcription-polymerase chain reaction only allow semiquantitative assessments and cannot accurately quantify the expression of the cytochrome P450 (CYP) enzyme. Therefore, the relationship between excess CYP enzyme and enzymatic activity is not been systematically studied yet. In the present study, we developed a new method using ultra-high-performance liquid chromatography (UPLC) mass spectroscopy (MS)/MS and determined the accurate quantification of CYP3A2 in rat liver. The linear range was 0.1329–8.508 ng/mL. The intra- and interday precisions and accuracies data fulfilled the acceptance criteria of food and drug association guidelines for bio analytical method validation. We compared the quantitative results with CYP3A2 activity obtained by the cocktail assay. The developed method was successfully used for the quantitative analysis of CYP3A2. It provided a good background for determining the differences in drug metabolism caused by different CYP expression. Keywords: UPLC-MS/MS; CYP3A2; Accurate Quantitation; Enzyme Activity

Validation of bioanalytical method for quantification of Vitamin K2 (MK-4) in human plasma by high-performance liquid chromatography-ultraviolet.

Vitamin K can reduce warfarin's anticoagulant action, causing a variance in response among individuals taking warfarin. Vitamin K comes in two forms, namely Vitamin K1 (phylloquinone) and K2 (menaquinones). Menaquinone-4 (MK-4) is a kind of Vitamin K2 found in meat and dairy products. Analysis of MK-4 levels in human plasma is very useful for patients who receive warfarin therapy. High-performance liquid chromatography (HPLC) can be used for warfarin's bioanalysis, and it must be validated. The purpose of this study was to validate the bioanalytical method for quantification of Vitamin K2 (MK-4) in human plasma according to the 2019 European Medicines Agency (EMA) guideline. Vitamin K2 (MK-4) was extracted using acetonitrile. HPLC with an ultraviolet detector at 245 nm, using a T3 column set at 30°C and an isocratic mobile phase containing methanol: phosphate buffer (95:5) at pH 3, a flow rate of 1 mL/min was used in this study. The warfarin concentration of 0.5-3 µg/mL was used. About 5.50%-17.42% and 6.18%-8.74%, respectively, were the average ranges of percentage coefficient of variation and percentage difference. There was no response at the analyte's retention time in the six blank plasmas and at the analyte's retention time in the blank after the injection of upper limit of quantification, indicates that the procedure was very selective and did not result in any carryover. This bioanalytical method fulfills the parameters of selectivity, accuracy, precision, and carryover based on the 2019 EMA guidelines.

Determination of a specific metabolite for the non-ionic surfactant 2,4,7,9-Tetramethyl-5-decyne-4,7-diol (TMDD) by UPLC-MS/MS

2,4,7,9-Tetramethyldec-5-yne-4,7-diol (TMDD) is a non-ionic surfactant commonly used as defoaming agent and numerous other applications. Effluents of wastewater treatment plants have been identified as one of the main sources of TMDD emissions into the environment. Due to its broad application in various fields, TMDD was selected for the development of a biomonitoring method for assessing human exposure within the frame of the cooperation project of the German Federal Ministry for the Environment, Nature Conservation, Building and Nuclear Safety (BMUB) and the German Chemical Industry Association (VCI) in 2020. This study aimed to identify a urinary metabolite for TMDD by UPLC-Q-Orbitrap-MS which can be used as a biomarker of TMDD exposure. Monohydroxylated TMDD (1-OH-TMDD) was deciphered as the most prominent metabolite of TMDD in humans in a series of in vitro and in vivo experiments.In a next step, a quantitative method for the determination of 1-OH-TMDD was developed and validated. Quantification was achieved by isotope dilution using D3-1-OH-TMDD as internal standard. The method is characterized by a simple sample clean-up procedure and an enzymatic hydrolysis of possible metabolite conjugates with ß-glucuronidase. Method validation was performed according to international guidelines for bioanalytical method validation. The method proved its robustness, precision, accuracy and sensitivity for the intended purpose, i.e. the assessment of TMDD exposure in the general population by means of human biomonitoring.

Bioanalysis of Endogenous Isotretinoin in Altered and Unaltered Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry Coupled with Electrospray Ionization.

Bioanalytical method development and validation of endogenous Isotretinoin with Isotretinoin D5 as internal standard was done as per current regulatory guidelines. The method is simple, rugged and sensitive enough to estimate endogenous Isotretinoin using the chromatography-tandem mass spectrometry technique. An alternative approach has been adopted for quantitative analysis of endogenous Isotretinoin in human plasma. Isotretinoin free matrix (surrogate matrix) was prepared and further used for the development and validation of Isotretinoin. The method was validated in altered and unaltered plasma. The chromatographic optimization was done with column (ACE C18, 100 × 4.6mm I.D. 5μm particle size), using a mobile phase containing 1mM ammonium acetate, pH3.0 as a solvent A and solvent B (1mM ammonium acetate (pH3.0) with acetonitrile in a ratio of 10:90). A flow rate was set at 0.75mL/min in a binary gradient mode. The analyte was recovered by liquid-liquid extraction method with diethyl ether as an extraction solvent. Multi-reaction monitoring mode in negative polarity was implemented for the quantification of endogenous Isotretinoin in plasma. The calibration curve of Isotretinoin was linear (r2 > 0.9992) over the concentration range of 0.5-1000ng/mL. The intra-day precision was found in a range of 2.0-3.9% CV for altered samples and 0.9-3.7% CV for unaltered samples. The inter-day precision was found 2.6-6.1% CV for altered samples and 1.3-3.8% CV for unaltered samples. The average recovery of the extraction procedure was found 64.6% for altered samples and 62.2% for unaltered samples.

Formulation development and optimization of taste-masked azithromycin oral suspension with ion exchange resins: Bioanalytical method development and validation, in vivo bioequivalence study, and in-silico PBPK modeling for the paediatric population

The study is based on preparing taste-masked oral suspension of azithromycin using ion exchange resins Kyron T135 and Doshion-P542 AB. The complexation process was optimized through Design of Experiment (DOE), and the resins-complexes were characterized by bitterness score, differential scanning calorimetry (DSC), fourier transform infra-red spectrophotometry (FTIR) and powder x-ray diffraction (PXRD). The complexes were further formulated and optimized through Central Composite Design (CCRD) into oral suspension using xanthan gum, hydroxypropyl cellulose, tri-basic sodium phosphate, and sucrose. To obtain the optimized product, the trial batches were tested for sedimentation ratio and viscosity in comparison to the reference product followed by assay and studies of drug-release kinetics. To perform bioequivalence of the optimized formulations against the reference product, an LC-MS/MS based bio-analytical method was developed and validated followed by a three-period cross over study. The pharmacokinetic parameters Cmax, Tmax, AUC0-t and AUC 0-∞ were obtained through non-compartmental analysis (NCA) using log-transformed data. Moreover, physiologically based pharmacokinetic (PBPK) modelling of the in-vitro data was carried out to predict the pharmacokinetics of the optimized formulations in the intended paediatric population. Kyron T-135 yielded the best loading efficiency at a drug resin ratio of 1:1.35 and Doshion-P542 AB at 1:3.25, while stirring and swelling time of 30 min for Kyron T-135 yielded optimum drug loading. In case of Doshion P-542 AB the optimum time was 22.5 and 30 min for stirring and swelling respectively. The bitterness score proved the tastelessness of the drug resin complex. The optimized formulation with Kyron (formulation code K4) and Doshion (formulation code D6) were selected as optimum on the basis of similar quality attributes to the reference product. The drug dissolution kinetics revealed first order drug release. The developed bioanalytical method was found to be sensitive and linear in the range of 2–500 ng/mL showing accuracy within 101.32–106.68%. Both optimized formulations (K4 and D6) were found to be bioequivalent with geometric mean ratio of Cmax, Tmax, AUC0-t and AUC 0-∞ within 90% C.I. in silico PBPK based predictive pharmacokinetics in the paediatric population was successfully estimated for the developed formulations.

A High-Throughput Fast Chromatography-Tandem Mass Spectrometry-Based Method for Deoxynivalenol Quantification in Wheat Grain

Fusarium head blight (FHB), caused by Fusarium spp., is a destructive disease of cereal grains. Apart from grain yield loss, a major quality concern is contamination with Fusarium-produced mycotoxins, specifically deoxynivalenol (DON). Mycotoxins accumulate in the grain, making it unfit for consumption by humans and animals. Breeding cultivars with high disease resistance and low mycotoxin contamination is a priority for wheat breeders. However, DON measurement in breeding programs is expensive and time consuming due to the lack of efficient quantification methods. In this study, we established a simple fast chromatography-tandem mass spectrometry method, which employed a one-step acetonitrile extraction protocol with a short guard column to reduce complexity, cost, and analysis time. To ensure robustness and reproducibility, the method was validated according to the U.S. Food and Drug Administration Guidance for Bioanalytical Method Validation. Furthermore, the method was applied for determination of DON in 102 wheat grain samples. Obtained results highly correlated with the conventional immunological method for all tested samples. With its ease of use, rapid sample analysis, and high sensitivity and accuracy, the method could be integrated into current FHB breeding programs to increase breeding efficiency and accelerate screening progress to identify germplasm with increased resistance to DON accumulation. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Rapid quantification of phenobarbital and barbital in human whole blood by liquid-liquid extraction combined with DART-orbitrap-HRMS.

This study aims to develop and validate a rapid, simple, and efficient bioanalytical method for the simultaneous quantification of phenobarbital and barbital in human whole blood using liquid-liquid extraction combined with direct analysis in real time (DART) and high-resolution mass spectrometry (HRMS). Phenobarbital-d5 and aprobarbital were selected as internal standards (ISs) of phenobarbital and barbital, respectively. A mixed solvent of o-xylene and ethyl acetate at a ratio of 1:6 was used to extract analytes of interest and ISs from 100 μL of human whole blood samples. Phenobarbital and barbital were detected by DART-HRMS. The proposed method has been validated in accordance with United States Food and Drug Administration Guidelines for Bioanalytical Method Validation in terms of selectivity, linearity, accuracy, precision, matrix effect, recovery, stability, and dilution integrity. The lower limits of quantification (LLOQs) of phenobarbital and barbital were both 10ng/mL. The linearities were in the range of 10-1000ng/mL (R2 ≥ 0.99). The mean recovery values of phenobarbital and barbital were 99.7% and 88.1%, respectively. The interday and intraday precision values were less than 10.4%, and the interday and intraday accuracy values ranged from 87.6 to 106.7%. Furthermore, the validated method was applied to four cases of phenobarbital poisoning at the Shanghai Institute of Forensic Science. The developed and fully validated method enabled the simultaneous quantification of phenobarbital and barbital in human whole blood and was successfully applied to authentic cases.

Bioanalytical method validation, biopharmaceutical and pharmacokinetic evaluation of GSK-650394, a serum- and glucocorticoid-regulated kinase 1 inhibitor

GSK-650394 is an inhibitor of serum- and glucocorticoid-regulated kinase 1 that displays potency for treating cancer, hypertension, cardiovascular and neuronal diseases, such as Parkinson’s disease. However, the biopharmaceutical properties and pharmacokinetics of GSK-650394 have not been studied extensively. Also, there are currently no bioanalytical assays available for this new drug candidate. In this study, we developed a simple and sensitive liquid chromatography-tandem mass spectrometry method to quantify GSK-650394 in rat plasma and validated its selectivity, linearity, accuracy and precision, sensitivity, matrix effects, extraction recovery, and stability, following the United States Food and Drug Administration guidelines. In vitro studies showed the biopharmaceutical properties of GSK-650394, including its low solubility in water and simulated gastrointestinal fluids, passive transport in Caco-2 cell monolayers, high plasma protein binding, and primary metabolism by glucuronide conjugation in the small intestine and liver of rats. Following intravenous administration (2 mg/kg) to rats, GSK-650394 exhibited low total clearance (11.18 ± 1.28 mL/min/kg) and volume of distribution at steady-state (346.1 ± 120.6 mL/kg). Following oral administration (2, 5, and 10 mg/kg) to rats, GSK-650394 underwent enterohepatic circulation, with low bioavailability (∼9%). The insignificant difference in bioavailability among three oral doses suggests that GSK-650394 may follow linear pharmacokinetics up to an oral dose of 10 mg/kg. In addition, the total form of parent drug and glucuronide conjugate in rat plasma from three oral doses showed a much higher value of area under the plasma concentration versus time curve than the parent drug, indicating that the primary metabolism process of GSK-650394 was glucuronidation. Our findings suggest that the low oral bioavailability of GSK-650394 is associated with its low solubility, instability under acidic gastric conditions, and extensive glucuronidation metabolism.

A Volumetric Absorptive Microsampling UPLC-MS/MS Method for Simultaneous Quantification of Tacrolimus, Mycophenolic Acid and Creatinine in Whole Blood of Renal Transplant Recipients.

(1) Background: Continuous monitoring of tacrolimus (TAC), mycophenolic acid (MPA), and creatinine (Cre) after renal transplantation is vitally important. In this study, we developed a new method based on volumetric absorptive microsampling (VAMS) combined with Ultra Performance Liquid Chromatography−Tandem Mass Spectrometry (UPLC-MS/MS) to simultaneously quantify three analytes including TAC, MPA, and Cre in whole blood. (2) Methods: The VAMS-based UPLC-MS/MS assay used a shared extraction and a single injection to simultaneously quantify the included TAC, MPA, and Cre. Development and validation were carried out following the Food and Drug Administration and European Medicines Agency guidelines for the validation of bioanalytical methods. Moreover, clinical validation for the three analytes was performed in both dried blood spot (DBS) and VAMS. Furthermore, a willingness survey was conducted using the system usability scale (SUS) for renal transplant recipients. (3) Results: The assay was successfully validated for all analytes. No interference, carryover, or matrix effects were observed, and extraction recoveries and process efficiencies were >90.00%. Analysis was unaffected by hematocrit (0.20~0.60, L/L) and anticoagulants (EDTA-2K). Dried VAMS samples were stable for 7 days at ambient temperature and stable for at least 1 month at −20 °C. During clinical validation, the measured TAC, corrected MPA, and Cre concentrations of VAMS samples met the analytical standards (95.00%, 88.57%, and 92.50%). When more stringent clinical acceptance criteria were set, the results obtained by VAMS (90.00%, 71.43%, and 85.00%) better than DBS (77.50%, 62.86%, and 70.00%). Compared with DBS, the survey found that renal transplant recipients are more inclined to use VAMS. (4) Conclusions: A robust extraction and UPLC-MS/MS analysis method in VAMS tips was developed and fully validated for the simultaneous quantification of TAC, MPA, and Cre concentrations. This method provides analytical support for the one-sample remote monitoring of both immunosuppressive drug concentrations and renal function in allo-renal recipients. Based on the good consistency between this method and the routine detection of venous blood samples and higher patient satisfaction than DBS, we believe that VAMS sampling can be a better alternative to venous whole-blood sampling.