one step Morphine urine test Strip

Intended use

One step morphine is an easy to use one-step immunoassay intended for use in the qualitative analysis for morphine and its metabolites in human urine. It should not be used without proper supervision and is not intended for sale to the general public. The test provides only preliminary screening test results; a more specific alternate chemical method must be used in order to confirm the analytical result. Gas chromatography / mass spectrometry (gc/ms) is the preferred confirmatory method (1). Clinical considerations and professional judgment should be applied to any drug of abuse analysis, particularly when positive results are indicated (2,3).

Summary and explanation of the test

Opiates such as morphine, heroin, and codeine are derived from the resin of opium poppy. This class of drugs is known as the cns depressants. At therapeutic doses, opiates have an analgesic action, which reduces the severity of traumatic pain. Acute higher doses, as used by abusers or addicts produces euphoria and release from anxiety. Signs of physical dependence include depressed coordination, disrupted decision-making, decreased respiration, hypothermia, and coma. Tolerance develops to the analgesic and cns effects with prolonged use. Narcotic analgesics are used to maintain addicts. Withdrawal symptoms from these opiates are manifested by excitability, anxiety, insomnia, anorexia, diarrhea, and muscle/joint aches. Heroin is quickly metabolized to morphine. Thus, morphine and morphine glucuronide might both be found in the urine of a person who has taken only heroin. The body also changes codeine to morphine. Thus, a positive result obtained from the morphine test may indicate heroin, morphine (or its metabolite, morphine glucuronide), and / or codeine use (5,6). The morphine test is designed to perform at a threshold of 300 ng/ml as recommended by national institute on drugs of abuse (nida) as the cutoff for screening opiates (federal register, april 11, 1988). Positive results should be confirmed by an appropriately sensitive and specific methodology using a deferent chemical principle. If performed under conditions that provide sufficient sensitivity, hplc, gc and gc/ms are generally acceptable alternate methods of confirmation of the morphine test (2-4). While confirmation techniques other than gc/ms may be adequate for some drugs of abuse, gc/ms is generally accepted as a rigorous confirmation technique for all drugs, since it provides the best level of confidence in the results.

Principles

The morphine test is a one-step membrane immuno-chromatographic test (6) based on antigen/antibody complexation and is used for the analysis of morphine and its metabolite present in the test sample. The assay is based on the competition between the drug or drug metabolite in the sample and a drug conjugate immobilized on a porous membrane support for limited antibody sites on the colored microspheres. Test urine is delivered into a sample site located at one end of the porous membrane. When the drug is present in the urine test sample, the drug or drug metabolite competes for the limited antibody sites on the microspheres. When an adequate amount of drug is present attachment of the colored microspheres to the probe site on the membrane. Therefore, a positive urine sample will inhibit the formation of a precipitin line at the probe area. A reference or control line with a secondary antibody reaction is added to the membrane test to indicate that the test is viable. This control line should always be present before making any test interpretation. Normally, a negative urine sample will produce two colored lines (the formation of a visible precipitin at the test zone in addition to the control line), and a positive urine sample will show only one line (the control line).

Materials provided

Each morphine test is individually sealed in a foil pouch.

Each test contains a membrane strip with a defined amount of colloidal gold conjugated with monoclonal (mouse) or polyclonal antibodies (sheep) in dry chemical form as absorbed in pad. A secondary polyclonal antibody (goat anti-mouse lgg) is used to form the reference line. Also, an antigenic drug conjugate an animal serum are adsorbed onto the membrane. The entire membrane is then dried before assembly and is used in the dry form. All necessary reagents for performing the test are preformulated and included in each individual test. No additional reagents are required. One needs only to add the test sample.

Each pouch contains:

1.one morphine test

2.desiccant

3.urine dropper (for cassette)

Material required but not supplied:

Timer, sample container and disposable gloves. No other equipment or reagents are needed.

Storage and stability:

The components of this kit are stable until the marked expiration date when stored in the sealed pouch at 2 degree-30 degree (36-86 fahrenheit).

· The test must be brought to room temperature prior to use. Do not allow test to be subjected to below freezing or above 30 degree temperatures.

· Urine samples if not used immediately after collection, should be stored refrigerated (2-8 degree).

· For storage longer than 3 days, the samples should be frozen (<-20 degree).

Warnings and precautions

Avoid cross-contamination of urine samples by using a new urine specimen cup and dropper for each urine sample.

2. If a test foil bag is torn or perforated, the test should not be used.

3. Do not use the assay beyond the expiration date as indicated on the pouch.

4. Open sealed pouch containing the test immediately prior to use. Prolonged exposure to ambient humidity will cause product deterioration.

5. Urine samples should not exhibit a strong positive test for protein since either heavy proteinuria or hematuria may adversely affect the performance of the test.

Warning: urine specimens and all materials coming in contact with them should be handled and disposed of as if infectious and transmitting infection. Avoid contact with skin.

Sample collection and preparation

The morphine test is formulated for use with urine specimens. Fresh urine specimens should be used, and the urine does not require any special pretreatment. Urine samples should be collected such that testing may be performed as soon as possible, preferably during the same day as specimen collection. The sample should be free from gross debris. Samples taking out from a refrigerated storage must be allow to warm up to room temperature before testing. Allow at least 30 minutes when the sample was first taken out from a refrigerated storage at 2-8 degree, give longer time if the sample needs to be thawed out first from frozen storage.

Test procedure

1. Bring the test components to room temperature before opening the pouch.

2. Remove the test dipstick from the foil wrapper by tearing along the “notch”.

3. Insert the test dipstick into the urine sample being careful not to exceed the maximum level indicated by the arrows.

4. Read the test result at five minutes.

Interpretation of the results

1. Positive. One pink-rose band appears in the control zone and no band appears in the test zone. A positive result indicates the morphine level is 300ng/ml or higher.

2. Negative. Two pink-rose color bands appear, one in the control region and one in the test region. A negative result indicates that morphine level is below the detection sensitivity of 300ng/ml.

3. Invalid. If there are no distinct color bands visible in both the test zone and the control zone or if there is a visible band in the test zone but not in the control zone, then the test is invalid. In this instance, retesting of the specimen is recommended.

Note: in order to prevent any incorrect results, the test results should not be interpreted after five minutes.

If the concentration of drugs in sample is close to cutoff value of the test, the test band may be indistinct. Standard samples with such results can be thought the indistincted line of cutoff value of 150ng/ml. If the test band does not appear, the result is can be judged positive.

Limitations of the procedure

The assay is designed for use with human urine only. A positive result from the opiates test only indicates the presence of opiates, morphine, or morphine glucuronide and does not indicate or measure intoxication. There is the possibility that other substances and/or factors not listed (e.g. Technical or procedural errors) may interfere with the test and cause false result. See specificity for lists of substances that will produce positive results and substances that do not interfere with test performance.

Performance characteristics

Study protocal for morphine sensitivity test

The cutoff level is 1000ng/ml. Standard drug (+ morphine (300)) was added to normal drug-free urine to achieve concentrations at 0 ng/ml (negative control), 150 ng/ml(50% below cutoff level), 225 ng/ml (25% below cutoff level), 300 ng/ml (cutoff level) and 375 ng/ml (50% above cutoff level). Ten (10) samples at each concentration were tested.

Interpretation of result

If two line appear, indicate negative result, using “-“ mark to record the result.

If one line appear, indicate positive result, using “+” mark to record the result.

All the results read at 5 minutes.

Sensitivity test results

Samples	1	2	3	4	5	6	7	8	9	10
Concentration(ng/ml)
0	-	-	-	-	-	-	-	-	-	-
150	-	-	-	-	-	-	-	-	-	-
225	-	-	-	-	-	-	-	-	-	-
300	+	+	+	+	+	+	+	+	+	+
375	+	+	+	+	+	+	+	+	+	+

All 10 samples at concentration of 300 ng/ml or above had positive results.

Cross reactivity/specificity

The following compounds, related and unrelated to morphine, were tested on wh accutm morphine. All compounds were prepared in drug-free normal urine.

Related compounds result

Codeine positive at 300 ng/ml

Hydromorphone positive at 300 ng/ml

Oxycodone negative at 100 mcg/ml

Morphine sulfate positive at 300 ng/ml

Morphine-3-b-d-glucuronide positive at 500 ng/ml\

Morphine-6-b-d-glucuronide positive at 500 ng/ml

Methadone negative at 100 mcg/ml

Nalorphine negative at 100 mcg/ml

Heroin positive at 300 ng/ml

Ethylmorphine positive at 300 ng/ml