Antech GLP - Specimen Requirements & Collection

Testing Volume Requirements

Hematology | Coagulation | Urinalysis | Clinical Chemistry
Special Chemistry | Immunoassays
Sample Collection

Hematology

500 uL EDTA Whole blood + slide made at site. Do not freeze!

Complete Blood Count
  Red Cell Count
  Hemoglobin
   Hematocrit
   Mean Corpuscular Volume (MCV)
   Mean Corpuscular Hemoglobin (MCH)
   Mean Corpuscular Hemoglobin Concentration (MCHC)
  Platelet Count
  Leukocyte Count
Leukocyte Differential
RBC Morphology

Reticulocyte Count

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500uL of whole blood or special stained cells, 2 slides

Coagulation

Prothrombin Time (PT)

Activated Partial Prothrombin Time (APTT)

Fibrinogen

Fibrin Degredation Products (FDP)

D-Dimer

500 µL frozen citrate plasma¹

500 µL frozen citrate plasma¹

500 µL frozen citrate plasma¹

500 µL frozen citrate plasma¹

500 µL frozen citrate plasma¹

¹ Sample must be collected in tube with ratio of 1 part sodium Citrate and 9 parts blood. Special tubes must be made for small volumes.

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Urinalysis

2 mL or entire specimen of urine refrigerated unspun. Provide total volume. Preferred collected over a timed period. Provide time period.

Appearance
Color
Osmolarity
Specific Gravity
Bilirubin*
Blood*
Glucose*

Ketones*
pH*
Protein* (May not be accurate in many species)
Urobilinogen*
Casts
Cells
Crystals

*Assays performed as semiquantitative dipstick method

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Clinical Chemistry

Minimum sample for one test 150 µL. Up to 20 tests can be performed on 350 µL of frozen serum. Separate from cells within 1 hour of collection.

Albumin
Alkaline Phosphatase (ALP)
Alanine Aminotransferase (ALT)
Aspartate Aminotransferase (AST)
Amylase
Bilirubin, Direct
Bilirubin, Total
Calcium
Chloride
Cholesterol
Creatine Kinase (CK)
Creatinine

Gamma Glutamyl Transferase (GGT)
Glucose
Lactate Dehydrogenase (LD)
Lipase
Magnesium
Phosphorus
Potassium
Sodium
Triglycerides
Total Protein
Urea Nitrogen

Clinical Chemistry tests can be performed on urine samples as quantitative assays. Indirect Bilirubin is a calculated parameter and requires Total and Direct Bilirubin. (No additional sample volume.)
Globulin and A/G ratio are calculated parameters and require Total Protein and Albumin. (No additional sample volume.)

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Special Chemistry

Alkaline Phosphatase Isoenzymes

Angiotensin Converting Enzyme (ACE)

Apolipoprotein

Bile Acids

CK Isoenzymes

Cholinesterase

CH50 Total Complement

C3 Complement

C4 Complement

Ferritin

Free Fatty Acids

Free Hemoglobin

HDL Cholesterol

Haptoglobin

Immunoglobulin A

Immunoglobulin G

Immunoglobulin M

Iron

LD Isoenzymes

LDL Cholesterol

Lactic Acid

Myoglobin

NAG

5'-Nucleotidase

Phopholipids

Protein Electrophoresis

Sorbitol Dehydrogenase (SDH)

Unsaturated Iron Binding capacity (UIBC)

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500 µL frozen serum

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 15 µL pipetted

500 µL serum preferred, 250 µL minimum

200 µL serum preferred, 100 µL minimum, 50 µL pipetted
500 µL of EDTA whole blood

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 20 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL Heparinized plasma preferred, 100 µL minimum, 25 µL pipetted. No vaccum tubes, use syringe.

200 µL serum preferred, 100 µL minimum, 2 µL pipetted

200 µL serum preferred, 100 µL minimum, 50 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 12 µL pipetted

500 µL serum room temperature, ship immediately, do not freeze.

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL Fluoride Plasma,
Whole blood–Prepare 1 part blood and 1 part 8% percholic acid precipitate. Freeze supernatant.

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL urine unpreserved preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 5 µL pipetted

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

500 µL serum

200 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 8 µL pipetted

Immunoassays

Adrenocorticotropic Hormone (ACTH)

Aldosterone

Antidiuretic Hormone (ADH)

Calcitonin

Catacholamines

Corticosterone

Cortisol

DHEA

DHEA Sulfate

Dihydrotestosterone

Estradiol

Folic Acid

Follical Stimulating Hormone (FSH)

Glucagon

Growth Hormone

Histamine

Insulin-Like growth Factor (IGF-1)

Insulin

Leutenizing hormone (LH)

Leptin

N-telopeptide

Osteocalcin

Parathyroid Hormone (PTH-intact)

Pro-BNP

Progesterone

Prolactin

Thryroid Stimulating Hormone (TSH)

Testosterone

Thyroxine (T4)

Free T4

Tri-iodothyronine (T3)

T3 Uptake

Troponin I

Vitamin B12

Vitamin D 1,25 OH

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200 µL EDTA plasma preferred, 100 µL minimum, 75 µL pipetted. Collect in cold siliconized tube, separate and freeze immediately.

500 µL serum preferred, 150 µL minimum, 100 µL pipetted

750 µL serum preferred, 500 µL minimum, 400 µL pipetted

Rat–500 µL of serum preferred, 250 µL minimum, 200 µL pipetted
Non-Rodents–500 µL of serum preferred, 200 µL minimum, 75 µL pipetted

500 µL serum preferred, 250 µL minimum, 200 µL pipetted

200 µL serum preferred, 150 µL minimum, 50 µL pipetted

200 µL serum preferred, 120 µL minimum, 10 µL pipetted

500 µL serum preferred, 150 µL minimum, 100 µL pipetted

500 µL serum preferred, 100 µL minimum, 25 µL pipetted

200 µL serum preferred, 100 µL minimum, 50 µL pipetted

500 µL serum preferred, 300 µL minimum, 250 µL pipetted

500 µL Heparinized Plasma preferred, 250 µL minimum, 200 µL pipetted
For Whole Blood–Record Hematocrit. 100 µL of blood in 2 mL of 1% ascorbic acid (fresh) Freeze immediately.

500 µL serum preferred, 150 µL minimum, 100 µL pipetted

500 µL EDTA plasma with 5000 U Trasylal/10 mL of blood, 250 µL minimum

500 µL serum preferred, 200 µL minimum, 100 µL pipetted

250 µL EDTA plasma, 100 µL minimum Freeze immediately.

Rat–100 µL serum preferred, 50 µL minimum, 10 µL pipetted.
NHP–100 µL serum preferred, 50 µL minimum, 20 µL pipetted.

100 µL serum preferred, EDTA or heparin acceptable, 50 µL minimum, 25 µL pipetted;

500 µL serum preferred, 150 µL minimum, 100 µL pipetted

500 µL serum preferred, 150 µL minimum, 100 µL pipetted

Serum–100 µL preferred, 75 µL minimum, 50 µL pipetted
Urine–500 µL preferred, 50 µL minimum, 25 µL pipetted

Rat–100 µL serum preferred (Heparin acceptable), 50 µL minimum, 20 µL pipetted.
NHP–100 µL serum preferred, 50 µL minimum, 10 µL pipetted

Rat–500 µL serum preferred, 250 µL minimum, 200 µL pipetted
Non-Rodent–200 µL serum preferred, 150 µL minimum, 50 µL pipetted

200 µL serum preferred, 150 µL minimum, 100 µL pipetted

200 µL serum preferred, 125 µL minimum, 25 µL pipetted
No gel seperators

500 µL serum preferred, 200 µL minimum, 100 µL pipetted
Canine–100 µL serum preferred, 50 µL minimum, 25 µL pipetted

500 µL serum preferred, 150 µL minimum, 100 µL pipetted
Canine–150 µL serum preferred, 125 µL minimum, 25 µL pipetted

150 µL serum preferred (Heparin acceptable), 120 µL minimum, 20 µL pipetted

125 µL serum preferred (Heparin acceptable), 115 µL minimum, 15 µL pipetted

500 µL serum preferred, 200 µL minimum

150 µL serum preferred (Heparin acceptable), 125 µL minimum, 25 µL pipetted

500 µL serum preferred, 200 µL minimum

175 µL serum preferred (Heparin acceptable), 150 µL minimum, 50 µL pipetted

250 µL serum preferred (Heparin acceptable), 200 µL minimum, 200 µL pipetted

500 µL serum preferred (Heparin acceptable), 250 µL minimum, 250 µL pipetted

Sample Collection

Sample collection is very important in the integrity and eventual interpretation of the data. There are many pre-analytical variables that can affect the results of clinical pathology testing. Collecting the correct sample collection tube, collection site, sample storage and shipment can all affect the result before analysis even begins.

For mice and rats, the typical blood collection procedures require that the animals are anesthetized and restrained. It is important to use the same collection methods that work best for your facility. Always use the same collection site as well(1,2). Values can vary from one collection site to another. For other animals as well as rats and mice, it is important in some cases to collect blood at a certain time of day depending on the circadian rhythm. Fasting may be important for some tests. Rats and mice are not truly fasting for almost 24 hours, although debates continue if rats should be fasted.

Please check the list below to determine the correct sample type (serum or plasma) for the test. Some anticoagulants are not acceptable for certain tests and will invalidate the test results. Plasma can be obtained from the use of several different anti-coagulants. Each type of anti-coagulant changes the plasma. Not all anticoagulants are acceptable for all tests.

Sample storage is also important. The test manufacturer has tested samples for the best storage conditions and the time that samples are stable at these conditions. If these conditions cannot be met, it is important to send the samples for analysis as soon as possible.

1. Neptun, D, Smith, C and Irons, R, Effects of Sampling Site and Collection Method on Variations in Baseline Clinical Pathology Parameters in Fischer-344 Rats, I Clinical Chemistry, Fundamental and Applied Toxicology, 5, 1180-1185 (1985).
2. Smith, C, Neptun, D and Irons, R, Effects of Sampling Site and Collection Method on Variations in Baseline Clinical Pathology Parameters in Fischer-344 Rats, II Hematology, Fundamental and Applied Toxicology, 7, 658-663 (1986).

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