LC-MS/MS Identification of a Bromelain Peptide Biomarker from Ananas comosus Merr

Table 1

Bromelain identity and quality control data.

Item profile	Specification		Result	Method
Botanical-pineapple	Ananas sp.		Ananas comosus	Visual
Plant parts used	Mature plant stem		Mature plant stem	Visual
Botanical extract	Bromelain		Bromelain	Maltodextrin diluent
Identification	Standard Match (Sigma)		Conforms to standard	HPLC*
Activity, potency	≥2400 GDU**/gram		2931 GDU/gram	GDU Assay
Solvent Residues	LOQ^‡ Limits USP
Methanol	1000 ppm	3000 ppm	<1000 ppm	Gas chromatography
Ethanol	1000 ppm	5000 ppm	<1000 ppm	Gas chromatography
Diethyl ether	2 ppm	5000 ppm	<2.0 ppm	Gas chromatography
Acetone	50 ppm	5000 ppm	<50 ppm	Gas chromatography
2-Propanol	200 ppm	5000 ppm	<1200 ppm	Gas chromatography
Dichloromethane	2 ppm	600 ppm	<2.0 ppm	Gas chromatography
n-Hexane (C6)	2 ppm	290 ppm	<2.0 ppm	Gas chromatography
Ethyl acetate	20 ppm	5000 ppm	<20.0 ppm	Gas chromatography
Xylenes (0, M, P, EB)	1 ppm	2170 ppm	≤1.0 ppm	Gas chromatography
Microbial Profile	(CFU/gm)		(CFU/gm)
Total mold and yeast	≤300/gram		160	MLP° USP 27
E. coli	Negative		Negative	MLP USP 27
Salmonella	Negative		Negative	MLP USP 27
Enterobacteriaceae	≤10 CFU/gram		<10/gram	MLP USP 27
S. aureus	Negative		Negative	MLP USP 27
P. aeruginosa	Negative		Negative	MLP USP 27
Aflatoxin Profile; (B1,B2,G1,G2)	Aflatoxins ≤20 ppb		Negative	HPLC
Heavy metal profile
Arsenic	≤3 ppm		<0.50 ± 0.5 ppm	ICP-MS!
Lead	≤10 ppm		0.08 ± 0.05 ppm	ICP-MS
Cadmium	≤3 ppm		<0.25 ± 0.25 ppm	ICP-MS
Mercury	≤2 ppm		<0.10 ± 0.1 ppm	ICP-MS

∗HPLC: high-performance liquid chromatography, **GDU’s: gelatin dissolving units; LOQ^‡: limit of quantification, °microbial limit tests; ! ICP-MS: inductively coupled plasma-mass spectrometry; ^#CDFA: California Department of Food and Agriculture.