Methods

The SMBC provides accurate and absolute quantitation of small molecules in biological samples. Our services include UPLC-MS/MS assay development and validation, and sample analysis with or without sample processing. Current methods available from the core are listed below.

Alpha Agonist Panel

Method validated for a panel of three alpha-2A adrenergic receptor agonists:

  • Clonidine (shown above), Lofexidine LLOQ 0.1 ng/mL
  • Tizanidine LLOQ 0.2 ng/mL

Typical sample volume: 100 μL human plasma
Typical Lower Limit of Quantitation (LLOQ): 0.1-0.2 ng/mL as shown above

Antiretroviral (ARV) Panel - Plasma

Method validated for a panel of nine antiretrovirals:

  • Abacavir (ABC)
  • Bictegravir (BIC)
  • Cabotegravir (CAB, shown above)
  • Dolutegravir (DTG)
  • Doravirine (DOR)
  • Emtricitabine (FTC)
  • Lamivudine (3TC)
  • Raltegravir (RAL)
  • Tenofovir (TFV)

Typical sample volume: 50 μL human plasma
Typical Lower Limit of Quantitation (LLOQ): 1 ng/mL

Antiretroviral (ARV) Panel - Tissue

Method validated for a panel of five antiretrovirals in cervical, rectal, and vaginal tissues:

  • Bictegravir (BIC)
  • Cabotegravir (CAB, shown above)
  • Dolutegravir (DTG)
  • Doravirine (DOR)
  • Raltegravir (RAL)

Typical sample volume: 0.5 mL tissue homogenate
Typical Lower Limit of Quantitation (LLOQ): 0.05 ng/mL

Asymmetric Dimethylarginine (ADMA)

Method validated for asymmetric dimethylarginine (ADMA, shown below): 

Typical sample volume: 0.2 mL cerebrospinal fluid (CSF); 0.1 mL serum
Typical Lower Limit of Quantitation (LLOQ): 1 ng/mL in CSF

Brimonidine and Timolol

Method validated for brimonidine and timolol (shown below):


Typical sample volume: 50 μL human serum
Typical Lower Limit of Quantitation (LLOQ): 0.05 ng/mL

Creatine and Guanidinoacetic acid

Method validated for creatine and guanidinoacetic acid (shown below):

Typical sample volume: 50 μL human plasma
Typical Lower Limit of Quantitation (LLOQ): 50 ng/mL

Endocrine Panels

Methods validated for selected compounds in three endocrine panels:

Estrogens:   

  • Estradiol  (shown above) LLOQ 1 pg/mL
  • Estrone  LLOQ 1 pg/mL 
  • Ethinylestradiol  LLOQ 2.5 pg/mL

Typical sample volume: 0.5 mL serum/plasma 
Typical Lower Limit of Quantitation (LLOQ): 1-2.5 pg/mL as shown above

Androgens:   

  • Testosterone 
  • Androstenedione 

Typical sample volume: 0.5 mL serum/plasma 
Typical Lower Limit of Quantitation (LLOQ): 25 pg/mL

Progestins:   

  • Etonogestrel 
  • Levonorgestrel 
  • Medroxyprogesterone acetate 
  • Norethindrone 
  • Progesterone 

Typical sample volume: 0.5 mL serum/plasma 
Typical Lower Limit of Quantitation (LLOQ): 25 pg/mL

Fexofenadine and Olmesartan

Method validated for fexofenadine and olmesartan (shown below): 

 

Typical sample volume: 50 μL human serum  
Typical Lower Limit of Quantitation (LLOQ): 1 ng/mL

Guanosine triphosphate (GTP)

Method validated for guanosine triphosphate (GTP, shown below):

Typical sample volume: 50 μL cell lysate
Typical Lower Limit of Quantitation (LLOQ): 25 ng/mL

Glyburide

Method developed and preliminarily validated for glyburide (shown below): 

 

Typical sample volume: 200 μL serum;  400 μL brain tissue
Typical Lower Limit of Quantitation (LLOQ): 50 pg/mL

20-HETE

Method validated for 20-hydroxyeicosatetraenoic acid (20-HETE, shown below):

Typical sample volume: 1.0 mL human cerebrospinal fluid (CSF); 0.5 mL human plasma
Typical Lower Limit of Quantitation (LLOQ): 0.2 ng/mL in human CSF; 0.4 ng/mL in human plasma

Ketamine Panel

Method validated for ketamine (shown below) and its metabolites including norketamine , dehydronorketamine, 2S,6S hydroxynorketamine, and 2R,6R hydroxynorketamine:

Typical sample volume: 200 μL serum 
Typical Lower Limit of Quantitation (LLOQ): 0.1 ng/mL

Lenacapavir (LEN)

Method validated for lenacapavir (LEN, shown below):

Typical sample volume: 50 μL human plasma
Typical Lower Limit of Quantitation (LLOQ): 0.1 ng/mL

LY2886721

Method validated for LY2886721 (shown below):

Typical sample volume: 25 μL serum or brain tissue
Typical Lower Limit of Quantitation (LLOQ): 0.5 ng/mL

Melatonin and N-Acetylserotonin

A method for melatonin and N-acetylserotonin (shown below) has been preliminarily developed:

Typical sample volume: 500 μL serum
Typical Lower Limit of Quantitation (LLOQ): 1 ng/mL

Metformin

Method validated for metformin shown below):

Typical sample volume: 50 μL human plasma
Typical Lower Limit of Quantitation (LLOQ): 5 ng/mL

Methylnicotinamide (MNA)

Method validated for 1-methylnicotinamide (MNA, shown below), a methylated amide of nicotinic acid in human serum or urine:

Typical sample volume: 50 μL serum or urine
Typical Lower Limit of Quantitation (LLOQ): 1 ng/mL for serum; 0.5 μg/mL for urine

N6-Methyladenosine (m6A)

Method validated for N6-methyladenosine (m6A, shown below):

Typical sample volume: 50 μL cell lysate
Typical Lower Limit of Quantitation (LLOQ): 0.1 ng/mL 

N-Acetylaspartate

Method developed and preliminarily validated for N-acetylaspartate (shown below):

Typical sample volume: 20 μL cerebrospinal fluid (CSF)
Typical Lower Limit of quantitation (LLOQ): 0.5 μM 

Nucleoside Panel 

A method for adenosine 3',5'-cyclic monophosphate (cAMP, shown below) and other metabolites has been preliminarily developed:

Typical sample volume: 100 μL cell lysates, urine or tissue
Typical Lower Limit of Quantitation (LLOQ): 0.05 ng/mL

Prostaglandin Panel

 A method for selected metabolites of arachidonic acid in the cyclooxygenase pathways has been preliminarily developed: 

  • Prostaglandin A2 (PGA2)
  • Prostaglandin E2 (PGE2, shown above)
  • Prostaglandin F1alpha (PGF1alpha): 6-keto PGF1alpha
  • Prostaglandin F2alpha (PGF2alpha): 11-beta PGF2alpha
  • Prostaglandin J2 (PGJ2): 15-deoxy PGJ2, Delta-12 PGJ2

Typical sample volume: Rat brain cortical tissue; 0.5 mL human plasma
Typical Lower Limit of Quantitation (LLOQ): 0.1-0.2 ng/mL in rat brain cortical tissue; 0.4 ng/mL in human plasma

Rivaroxaban and Apixaban

Method validated for rivaroxaban and apixaban (shown below):



Typical sample volume: 100 μL serum
Typical Lower Limit of Quantitation (LLOQ): 1 ng/mL

Sedative Panel

Method validated for a panel of nine sedatives and metabolites:

  • Midazolam (MDZ) (shown above), morphine (MOR)  and  hydromorphone (HM) LLOQ 0.5 ng/mL
  • Alpha hydroxymidazolam (1- OH MDZ) (shown above),  morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) LLOQ 5 ng/mL
  • Dexmedetomidine (DEX), fentanyl (FEN) and norfentanyl (NF) LLOQ 0.05 ng/mL

Typical sample volume: 100 μL human plasma
Typical Lower Limit of Quantitation (LLOQ): 0.05-5 ng/mL as shown above

Sildenafil

Method validated for sildenafil and N-desmehyl sildenafil (shown below):



Typical sample volume: 50 μL serum, tissue, or amniotic fluid
Typical Lower Limit of Quantitation (LLOQ): 0.5 ng/mL

Tetrahydrocannabinol (THC) Panel

Method validated for tetrahydrocannabinol (THC, shown below), 11-hydroxy-THC, and 11-nor-9-carboxy-THC:

Typical sample volume: 150 µL human plasma; 150 µL human serum; 150 µL rat plasma
Typical Lower Limit of Quantitation (LLOQ): 0.25 ng/mL
TMAO Panel

Method validated for three metabolites of dietary phosphatidylcholine:

  • Trimethylamine N-oxide (TMAO, shown above) 
  • Choline 
  • Betaine 

Typical sample volume: 50 μL human plasma or urine
Typical Lower Limit of Quantitation (LLOQ): 10 ng/mL in plasma; 1000 ng/mL in urine

Uremic Toxin Panel

Method validated for four gut-derived uremic toxins:

  • Kynurenic acid  LLOQ 10 ng/mL
  • Hippuric acid  LLOQ 200 ng/mL
  • Indoxyl sulfate (shown above)  LLOQ 200 ng/mL
  • p-Cresol sulfate  LLOQ 250 ng/mL

Typical sample volume: 50 μL human serum 
Typical Lower Limit of Quantitation (LLOQ): 10-250 ng/mL as shown above

Vitamin D Metabolite Panel

Method validated for 25-hydroxy vitamin D2, 25-hydroxy vitamin D3 (shown below) and selected metabolites: 

Typical sample volume: 500 μL serum
Typical Lower Limit of Quantitation (LLOQ): 1 ng/mL

Warfarin and Metabolites

Method validated for R- and S-warfarin (shown below), warfarin and its metabolites:

R- and S-warfarin

Typical sample volume: 0.5 mL human urine 
Typical Lower Limit of Quantitation (LLOQ): 5 ng/mL

Warfarin and Alcohol Metabolites

Typical sample volume: 0.1 mL 10mM Tris-HCl incubation buffer
Typical Lower Limit of Quantitation (LLOQ): Warfarin 100 ng/mL; Warfarin Alcohols: 0.5 ng/mL

New UPLC-MS/MS assays are constantly being developed and validated. Please contact Dr. Raymond E. West III for more information.