© Springer-Verlag Berlin Heidelberg 2005 II.4.1 Diphenylmethane antihistaminics by Yoko Hieda and Kojiro Kimura Introduction Diphenylmethane antihistaminics are being widely used for t
Trang 1© Springer-Verlag Berlin Heidelberg 2005
II.4.1 Diphenylmethane
antihistaminics
by Yoko Hieda and Kojiro Kimura
Introduction
Diphenylmethane antihistaminics are being widely used for treatments of allergy, motion
and HPLC [7–13] In this chapter, a GC method for simultaneous analysis of diphenylmethane antihistaminics and also HPLC methods for some representative drugs of this group are pre-sented
Simultaneous analysis by GC [4]
Reagents and their preparation
citrate, orphenadrine hydrochloride, benactyzine hydrochloride, doxylamine succinate, carbinoxamine maleate, chlorpheniramine maleate, triprolidine hydrochloride, homo-chlorcyclizine dihydrochloride, hydroxyzine dihydrochloride, clemastine fumarate and meclizine dihydrochloride can be purchased from Sigma (St Louis, MO, USA) Pure
Kissei Pharmaceutical Co., Ltd., Nagano, Japan and Nippon Shinyaku Co., Ltd., Kyoto,
(Milford, MA, USA) Other common chemicals were of the highest purity commercially available
compounds (5-mg each as the weight of its free base) are altogether dissolved in methanol
whole blood or urine One of the 15 drugs is selected for use as internal standard (IS)
pre-pare 100 mL solution
GC conditions
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CA, USA); detector: FID; column temperatures: 160 °C (1 min) →5 °C/min →290 °C for the DB-1 column, and 160 °C (1 min) →5 °C/min →280 °C for the DB-17 column; injection
aliquot of sample solution is injected into GC in the splitless mode (1 min), followed by the split mode at 160 °C of oven temperature
Structures of principal diphenylmethane antihistaminics.
⊡ Figure 1.1
Trang 3well with 9 mL distilled water for complete hemolysis To this mixture, 5 mL of 0.5 M
are added
with a fl ow rate not faster than 5 mL/min using a 10-mL volume glass syringe
eluted with 3 mL of chloroform/methanol (9:1) into 4-mL volume glass vial
v A small amount of the upper aqueous phase of the eluate is carefully removed with a Pas-teur pipette; the lower organic phase is evaporated to dryness under a stream of nitrogen
For quantitation, the peak area ratio of a target compound to IS is obtained
target compound are added to 1 mL of blank whole blood or urine obtained from healthy subjects; at least 4 vials containing diff erent concentrations of the compound should be
obtained at the step v is applied to the calibration curve to obtain the concentration
Assessment and some comments on the method
> Figure 1.2 shows gas chromatograms for the authentic diphenylmethane antihistaminics
1 mL obtained by the present method using a DB-1 capillary column Many compounds ap-peared as sharp peaks With an intermediately polar DB-17 capillary column, sharp peaks also appeared By using both DB-1 and DB-17 columns, most compounds can be separated with sharp peaks; however phenyltoloxamine and orphenadrine (peaks 3 and 4) could not be sepa-rated with either column With the DB-1 column, the peak of triprolidine (peak 9) overlapped
an impurity peak of whole blood and urine extracts, and the peak of chlorpheniramine (peak
over-come by using the DB-17 column
not lower than 90 % except benactyzine, piperilate and meclizine; those of the latter drugs were 64.6, 72.2 and 79.8 %, respectively
most compounds, molecular or quasi-molecular peaks are missing in the positive EI mode [4]
Trang 4318 Diphenylmethane antihistaminics
Trang 5except for terolidine and triprolidine For the latter compounds, relatively intense molecular ions appear to be used for sensitive detection In the positive and negative CI modes, intense quasi-molecular ions appear for most drugs, which can be used for sensitive quantitation
HPLC analysis of diphenyhydramine in blood and urine [7]
Diphenhydramine is one of the most popular drugs in this group, and its poisoning cases are many In this section, one of the most common method by HPLC for diphenyldramine is described
Reagents and their preparation
Sigma
solu-tion
aque-ous solution and 0.5 % (v/v) phosphoric acid solution
HPLC conditions
An HPLC system includes a usual conveying pump, an injector and a UV detector Column:
phase: acetonitrile/distilled water/1 M sodium dihydrogenphosphate solution (11:7:2, v/v); its
fl ow rate: 1.8 mL/min: detection wavelength: 205 nm
Procedure
car-bonate solution are added and extracted with hexane/isopropanol (98:2, v/v) by shaking
com-pounds An aliquot of the aqueous phase is injected into HPLC
Assessment of the method
could be obtained in the range of 1–100 ng/mL; the detection limit was reported to be 1 ng/mL
Trang 6320 Diphenylmethane antihistaminics
HPLC analysis of chlorpheniramine and its metabolites in blood and urine [8]
Chlorpheniramine is one of the most popular antihistaminics, and has been being used for over
50 years In this section, HPLC analysis of chlorpheniramine and its metabolites didemethyl-chlorpheniramine and demethyldidemethyl-chlorpheniramine is described
Reagents and their preparation
HPLC conditions
An HPLC system to be used includes a usual conveying pump, an injector and a UV detector
phase: acetonitrile/75 mM phosphate buff er solution (pH 2.5) (25:75, v/v); its fl ow rate: 2 mL/ min; detection wavelength: 254 nm
Procedure
added and extracted with 5 mL ethyl ether
to dryness
HPLC
Assessment of the method
chlorphe-niramine and IS under the conditions were 2.7, 3.2, 4.3 and 5.0 min Good linearity was found
in the range of 0–30 ng/mL; the detection limit was reported to be 1 ng/mL
HPLC analysis of hydroxyzine in blood [9]
Hydroxyzine has a similar structure to those of diphenylmethane antihistaminics, but is being widely used as an anxiolytic drug
Trang 7Reagents and preparation
solu-tions
HPLC conditions
An HPLC system includes a usual conveying pump, an injector and a UV detector Column:
phase: acetonitrile/75 mM phosphate buff er solution (pH 3.0, containing 20 mM
229 nm
Procedure
extracted with 5 mL of diethyl ether
injected into HPLC
Assessment of the method
Under the conditions, the peaks of IS and hydroxyzine appeared at 3.6 and 6.9 min, respec-tively Good linearity was observed in the range of 0–100 ng/mL; the detection limit was re-ported to be 3 ng/mL In this method, triprolidine is used as IS; therefore, triprolidine can be measured by this method using hydroxyzine as IS conversely
Poisoning cases, and toxic and fatal concentrations
appear aft er ingesting 3–5 times the therapeutic dose [14] Plasma diphenhydramine
Dimenhydrinate is a multi-component drug consisting of diphenhydramine (53–55 %)
fatal cases in which 20–40 mg/kg of dimenhydrinate is ingested by adults and not more than
500 mg ingested by infants In a fatal case of a victim, who had ingested 5 g dimenhydrinate,
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symptoms appeared for subjects whose urinary concentrations of diphenhydramine were not lower than 100 ng/mL [18] Another fatal case with dimenhydrinate showing 4.8 ng/mL of blood diphenhydramine was reported [19]
For hydroxyzine, fatal cases were reported with its blood concentrations at 1.1 [20] and
poisoning symptoms [23, 24]
antihistaminic action; since its structure is very similar to that of diphenhydramine, it is
A fatal case involving chlorpheniramine was reported [27] However, most of the poisoned
Notes
a) Any columns made of non-polar 100 % dimethylsilicone and intermediately polar 50 % phenylsilicone/50 % dimethylsilicone stationary phases can be used, regardless of their manufacturers
b) Any GC instrument for a capillary column can be used
be reused aft er passing urine and plasma specimens, but new ones cannot be reused With
may appear and interfere with the GC analysis according to a lot of the cartridges In such
a case, the conventional liquid-liquid extraction can be made in place of the solid-phase extraction Briefl y, aft er adding IS to a specimen, the solution is made alkaline by adding KOH or NaOH solution, followed by extraction with an organic solvent (diethyl ether or dichloromethane) [1]; the organic phase is back-extracted with acidic solution ( phosphoric
ex-traction with an organic solvent; the latter is condensed and subjected to GC analysis [2] According to a specimen, the fi rst organic extract can be directly used for GC analysis without the back-extraction When an organic extract is evaporated to dryness, the residue
is dissolved in a small amount of an organic solvent for GC analysis, or dissolved in a small amount of a mobile phase for HPLC analysis
d) As IS, one of other diphenylmethane antihistaminic, which shows a retention time close to
extraction procedure
e) For a whole blood, a total volume of the specimen solution to be poured into the cartridge
is 15 mL; 1 mL of a blood specimen is placed in a 50-mL volume beaker, followed by the
10-mL volume glass syringe, 7.5 mL of the solution is drawn into it and poured into the cartridge slowly; this procedure is repeated to apply all of the solution onto the cartridge f) It is used for preventing hydroxyzine from its adsorption to the HPLC column
Trang 91) Yoo SD, Axelson JE (1986) Determination of diphenhydramine in biological fluids by capillary gas chromatogra-phy using nitrogen-phosphorus detection Application to placental transfer studies in pregnant sheep J Chro-matogr 378:385–393
2) Blyden GT, Greenblatt DJ, Scavone JM et al (1986) Pharmacokinetics of diphenhydramine and demethylated metabolite following intravenous and oral administration J Clin Pharmacol 26:529–533
3) Hattori H, Yamamoto S, Iwata M et al (1992) Determination of diphenylmethane antihistaminic drugs and their analogues in body fluids by gas chromatography with surface ionization detection J Chromatogr 581:213–218 4) Seno H, Hattori H, Kumazawa T et al (1993) Positive- and negative-ion mass spectrometry of diphenylmethane antihistaminics and their analogues and rapid clean-up of them from biological samples Forensic Sci Int 62:187–208
5) Simons K, Singh M, Gillespie CA et al (1996) An investigation of the H1-receptor antagonist triprolidine : phar-macokinetics and antihistaminic effects J Allergy Clin Immunol 77:326–330
6) Nishikawa M, Seno H, Ishii A et al (1997) Simple analysis of diphenylmethane antihistaminics and their ana-logues in bodily fluids by headspace solid-phase microextraction-capillary gas chromatography J Chromatogr Sci 35:275–279
7) Selinger K, Prevost J, Hill HM (1990) High-performance liquid chromatography method for the determination
of diphenhydramine in human plasma J Chromatogr 526:597–602
8) Simons KJ, Simons FER, Luciuk GH et al (1984) Urinary excretion of chlorpheniramine and its metabolites in children J Pharm Sci 73:595–599
9) Simons FER, Simons KJ, Frith EM (1984) The pharmacokinetics and antihistaminic of the H1 receptor antagonist hydroxyzine J Allergy Clin Immunol 73:69–75
10) Simons KJ, Watson WTA, Chen XY et al (1989) Pharmacokinetic and pharmacodynamic studies on the H1-re-ceptor antagonist hydroxyzine in the elderly Clin Pharmacol Ther 45:9–14
11) Matsuda M, Mizuki Y, Terauchi Y (2001) Simultaneous determination of the histamine H1-receptor antagonist ebastine and its metabolites, carebastine and hydroxyebastine, in human plasma using high-performance liquid chromatography J Chromatogr B 757:173–179 and 765:205 (erratum)
12) Nishikawa M, Nakai A, Fushida H et al (1993) Enantioselective pharmacokinetics of homo-chlorcyclizine III Simultaneous determination of (+)- and (–)-homochlorcyclizine in human urine by high-performance liquid chromatography J Chromatogr 612:239–244
13) Sakurai E, Yamasaki S, Iizuka Y et al (1992) The optical resolution of racemic chlorpheniramine and its stereose-lective pharmacokinetics in rat plasma J Pharm Pharmacol 44:44–47
14) Sakamoto T (ed and translation) (1999) Poisoning Handbook Medical Science International Publication, Tokyo,
pp 62–64 (in Japanese)
15) Fucci N (1996) A case of lethal intoxication after ingestion of toquilone compositum Am J Forensic Med Pathol 17:231–232
16) Hausmann E, Wewer H, Wellhöner HH et al (1983) Lethal intoxication with diphenhydramine Report of a case with analytical follow-up Arch Toxicol 53:33–39
17) Karch SB (1998) Diphenhydramine toxicity: comparisons of postmortem findings in diphenhydramine-, cocaine-, and heroin-related deaths Am J Forensic Med Pathol 19:143–147
18) Winn RE, McDonnell KP (1993) Fatality secondary to massive overdoses of dimenhydrinate Ann Emerg Med 22:1481–1484
19) Farrell M, Heinrichs M, Tilelli JA (1991) Response of life threatening dimenhydrinate intoxication to sodium bicarbonate administration J Toxicol Clin Toxicol 29:527–535
20) Spiehler VR, Fukumoto RI (1984) Another fatal case involving hydroxyzine J Anal Toxicol 8:242–243
21) Johnson GR (1984) A fatal case involving hydroxyzine J Anal Toxicol 6:69–70
22) Magera BE, Betlach CJ, Sweatt AP et al (1981) Hydroxyzine intoxication in a 13-month-old child Pediatrics 67:280–283
23) Koyama K, Kikuno T, Kagami H et al (2000) Blood concentrations and symptoms in acute hydroxyzine poison ing cases J Nippon Hosp Pharm Assoc 26:13–16 (in Japanese)
24) Uges DRA (1997) Blood level data In: Brandenberger H, Maes RAA (eds) Analytical Toxicology for Clinical, Forensic and Pharmaceutical Chemists Walter de Gruyter, Berlin, pp 707–718
25) VanHarreweghe I, Mertens K, Maes V et al (1999) Orphenadrine poisoning in a child: clinical and analytical
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26) DeMercurio D, Chiarotti M, Giusti GV (1979) Lethal orphenadrine intoxication: report of a case Z Rechtsmed 82:349–353
27) Read D (1981) A fatal case involving chlorpheniramine J Toxicol Clin Toxicol 18:941–943
28) Winek CL (1977) Injury by chemical agents In: Tedeschi CG, Eckert WG, Tedeschi LG (eds) Forensic Medicine, Vol III Saunders, Philadelphia, pp 1568–1587