Analytical Laboratory Shared Resource

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Cancer Research Center of Hawaii

Analytical Laboratory Shared Resource
Faculty Director: Adrian Franke, Ph.D.

This laboratory was established to provide a central shared resource for the analysis of molecules of interest from human tissues and body fluids in support of clinical, epidemiologic and basic science studies relating to the prevention and understanding of cancer in human populations. In this capacity, it provides a key link in facilitating collaborative interdisciplinary research at the CRCH and utilizes results from Dr. Adrian Franke’s independent research activities in aspects of cancer prevention.
Under the direction of Dr. Franke, this laboratory is conducting analyses of clinical analytes (homocysteine, triglycerides, HDL- and total cholesterol, creatinine, and others) and lipid soluble plasma antioxidants (trans-lutein, trans-zeaxanthin, cis-lutein, cis-zeaxanthin, trans-anhydrolutein, cis-anhydrolutein, total a-cryptoxanthin, trans-b-cryptoxanthin, cis-b-cryptoxanthin, total lycopene, total dihydrolycopene, total a-carotene, trans-b-carotene, cis-b-carotene, retinol, retinyl palmitate, a-, b-, g-, and d-tocopherol) in support of several NIH-funded projects, including research on colorectal cancer (L. Le Marchand), lung cancer (L. Le Marchand), breast cancer (M. Goodman, A.Franke/Q. Dai), cervical cancer (M. Goodman), bladder cancer (A. Nomura) and prostate cancer (L. Kolonel). It also provides measurement of serum carotenoids and vitamins for projects evaluating intervention studies (G. Maskarinec) and determines these analytes in tissue and cells (J.Bertram, M. Goodman, B, Hernandez). ). Assays are updated continuously using state-of-the art technology. For example, blood Vitamin C is now measured by liquid chromatography with electrochemical detection resulting in faster and more accurate analyses relative to the traditional colorimetric method. Most HPLC based assays have been or are being changed towards smaller column diameters resulting in higher sensitivity, faster analysis time, and less waste generation.
CYP and NAT phenotyping analyses continue to be performed by determining caffeine and its metabolites, acetyl-formyl-methyl-uracil, 1,7-dimethylxanthin, and 1,7-dimethyl urate, in urine for studies on colorectal cancer susceptibility in adenoma patients (L. Le Marchand). Urinary isoflavonoid (daidzein, glycitein, genistein, dihydrodaidzein, dihydrogenistein, equol, and O-desmethylangolensin) and lignan (enterodiol, enterolactone) analyses are being performed to support research on the evaluation of potential cancer preventive effects of soy consumption against breast cancer (G. Maskarinec, A. Franke/W. Zheng) and other cancers (M. Goodman, B. Hernandez, A. Nomura). This laboratory determines micronutrient levels including carotenoids, tocopherols, tocotrienols, isoflavonoids and flavonoids (hesperetin, naringenin, apigenin luteolin, kämpferol, quercetin, myricetin, pelargonidin, cyanidin, and delphinidin) of various food items for the Nutrition Support Shared Resource food composition database (S. Murphy) and for other newly funded research projects.
In addition to HPLC instrumentation mostly used with photo-diode array, electrochemical, and fluorescence detection, this laboratory applies assays based on liquid chromatography mass spectrometry (LC/MS) and gas chromatography mass spectrometry (GC/MS) , which were funded by an NCI Cancer Center Support Grant supplement. LC/MS and GC/MS assays offer improved sensitivity, selectivity and accuracy of all the above measurements. New LC/MS assays are currently implemented for protein analysis, CYP 3A4 phenotyping (cortisol, 6-beta-hydroxy cortisol), sex hormones, various natural products (T. Hemscheidt, R. Moore, K.Czisar), and many other analytes as required by Center members.

SERVICES OFFERED
Analysis:

Total cholesterol in plasma by AA
Triglycerides in plasma by AA**
Lipid panel (Chol, HDLC, TGL) in plasma by AA
APO-A1 in plasma by AA
Apo-B in plasma by AA
Apo-E in plasma by AA
Haptoglobin in plasma by AA
Vitamin C in plasma, food by HPLC/ECD or PDA
LPM in plasma, foods by HPLC/PDA
Tocopherols in plasma by NP-HPLC
Caffeine and its metabolites in urine
DNA by HPLC
Total phenol levels in food, urine, plasma
Creatinine in urine by AA
CRP in plasma by AA
Calcium in plasma by AA
Ammonia in urine
Urea in urine
HDL-cholesterol in plasma by AA
Homocysteine in plasma
Fatty acids in plasma or RBC's
Red blood cell preparation
1-hydroxy-pyrene in urine
Isoflavonoids, flavonoids, lignans, anthocyanins or other groups of phenolic agents from plasma, urine, breast milk, foods.
Plasma or serum conzyme Q10 by HPLC/PDA
Consultation

technical (staff)
professional (director)
lab experiments (total analysis including supplies)

Enzyme immunoassays

Leptin
25 Hydroxy Vitamin D
Adiponectin
Interleukin-2
Interleuken-6 soluble Receptor
TNF alpha Receptor I
TNF alpha Receptor II
TNF alpha
Parathyroid Hormone
Urinary Isoprostane
Interleukin-6

Equipment use

HPLC/PDA systems (no supplies)
LC/MS system (no supplies)

AA = clinical auto analyzer
ECD = electrochemical detection
HPLC = high pressure liquid chromatography
LC/MS = liquid chromatography mass spectrometry
LPM = Lipid-Phase Micronutrients (12 carotenoids, 2 retinoids, 3 tocopherols)
PDA = photo diode array detection
RBC = red blood cells