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Detection of Morphine and Morphine-Glucuronide in Saliva or Urine with a Rapid and Simple Method Without Derivatization on the Varian 500-MS

ls were reagent or HPLC grade from Sigma-Aldrich (St. Louis, MO) with the exception of the morphine and morphine-glucuronide (Cerilliant, Little Rock, TX). Drugs of abuse urine control was obtained from Utak Laboratories, Inc (Valencia, CA). OnTrak OraTube was obtained from the local Varian sales office.

Sample Preparation

All stock solutions were prepared in 50:50 Methanol:Water at 1 mg/mL. All dilutions were prepared in 5% aqueous methanol + 0.1% acetic acid. Varian OnTrak OraTube (Cat# 6902051) (for obtaining a saliva sample) was used according to its instructions. Utak Laboratories Drugs of Abuse Confirmation Standard (Cat# 98815) was prepared, according to its instructions, by adding 10 mL of HPLC grade water to the dried samples provided.

All saliva and urine samples were diluted 1:10 with 5% aqueous methanol containing 0.1% acetic acid before injection on the 500-MS.

Results and Discussion

The structures of morphine and one of its metabolites are shown in Figure 1. Glucuronidation (catalyzed by uridine-glucuronic transferase or UGTs) is a Phase II metabolism reaction which involves the addition of glucuronic acid from uridine-diphosphoglucuronide to the xenobiotic being metabolized. In general, morphine metabolism is undertaken by UGT2B7.

Typically, simple onsite kits such as the Varian OnTrak TesTcard perform preliminary screening for morphine usage, either on a random basis or as an incident-driven event. However, to eliminate false positives, a second, but more specific chemical method, such as GC/MS, is used as a confirmatory test. However, GC/MS methods for drugs-of-abuse testing are time-consuming, requiring the use of solid-phase extraction and derivatization with pentafluoropropionic anhydride/ pentafluoropropanol. The LC/MS method uses a fast gradient program for complete analysis
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