Development and Validation of Analytical Method for Simultaneous Estimation of Bupivacaine and Meloxicam in Human Plasma Using UPLC-MS/MS

Abstract

Bupivacaine HCl (BP) and Meloxicam (ME) are used as intra-operative local anesthesia and non-steroidal anti-inflammatory drugs respectively. A simple, rapid, selective and sensitive ultra pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for simultaneous quantification of BP and ME in human plasma. By following a simple protein precipitation with solvent mixture containing Water:Acetonitrile:Formic Acid (76:24:0.1,v/v/v), the analytes and internal standards were separated on Acquity HSS T3 column (2.1 x 50 mm, 1.8 µm). Chromatographic Mobile phase A consist of 10 mM Ammonium Formate and Mobile phase B consist of a mixture of Acetonitrile:water:Formic acid (96:5:0.2, v/v/v). Bupivacaine-d 9 (BUd 9) used as internal standard for BP and 13 C,d 3 Meloxicam (ME 13 cd 3) used for ME. The concentration of BP and ME were determined by gradient UPLC-MS/MS in Multiple reaction mode (MRM). The mass transition ion-pair for BP was m/z 289.3→140.0 and BUd 9 was 298.3→ 149.0, for ME was m/z 352.1→115.0 and ME 13 cd 3 was 356.1→115.0, respectively. The proposed method was linear in the range 10-4500 ng/mL. The intra and inter-run precision values are within 6.8 % at a lower limit of quantization level. The overall recovery for both analytes found 98%. The total run time of the method is 2.5 min.

Determination of bupivacaine tissue concentration in human biopsy samples using high‐performance liquid chromatography with mass spectrometry

“…Repeated freezing and thawing for three times did not affect the stability of local anesthetics. The results are congruent with the literature where bupivacaine and lidocaine proved to be very stable in tissues and in different solvents (Cho et al, 2017; Golub et al, 1998; Scherpenisse & Bergwerff, 2007; Siddareddy et al, 2017).…”

“…For acid extraction (Cho et al, 2017; Siddareddy et al, 2017), tissue samples with bupivacaine and lidocaine were first homogenized in acidic acetonitrile (1% formic acid in acetonitrile). Then tissue was removed by centrifugation and the supernatant transferred into a fresh tube.…”

“…Several methods for measuring bupivacaine and other local anesthetics concentration in liquid samples of human and animal urine (Daryanavard et al, 2013; Lindberg, Kanto, & Pihlajamäki, 1986; Lombardo‐Agüí, Cruces‐Blanco, & García‐Campaña, 2009; Ma, Kang, Zhao, Chen, & Yao, 2006), plasma or serum (Gross, Nicolay, & Eschalier, 1999; Lindberg et al, 1986; Lindberg & Pihlajamäki, 1984; Siddareddy, Reddy, Suresh, & Sreeramulu, 2017), synovial fluid (Gough, Mayhew, & Munroe, 2002; Jordana et al, 2016) and microdialysis samples (McDonald, Faibushevich, Garnick, Mclaughlin, & Lunte, 2002) have been successfully developed. However, to date, only a few studies have attempted to determine the concentrations of local anesthetics in animal tissues.…”

Determination of drugs in exhumed liver and brain tissue after over 9 years of burial by liquid chromatography–tandem mass spectrometry—Part 2: Benzodiazepines, opioids, and further drugs

“…Another factor influencing detectability may be the light‐sensitivity of a drug leading to losses during sample preparation and exhumation 1 . Bupivacaine is light‐sensitive, which may have caused its detectability of only 67% (cases within time frame) 40 . However, stability of drugs in “alkaline medium and towards bacterial activities” is relevant during decomposition 2 .…”

“…(cases within time frame). 40 However, stability of drugs in "alkaline medium and towards bacterial activities" is relevant during decomposition. 2 This may be one reason for the negative results of butylscopolamine in all ( 6) analyzed cases as it is extensively decomposed in alkaline medium.…”