PROPERTIES AT A GLANCE

-Zivak LC-MS/MS Organic acid analysis kit is designed and produced for the screening of organic acids excreted in urine for the detection and diagnosis of inborn errors of metabolism and organic acidemias.

-Zivak LC-MS/MS Organic acid analysis kit is designed for the highly sensitive and specific determination of organic acids within a single analytical run.

-The organic acid determinations have a very high specificity of 100 %.

-The organic acid determinations have a high analytical sensitivity (LOD, 0,05 — 0,3 mg/L; LOQ, 0,1- 1,0 mg/L).

-The method is very suitable for clinical laboratory settings owing to a single step sample preparation and a 16 minute run time. 30 samples can be analyzed within a day (8 hours).

-Zivak LC-MS/MS Organic acid analysis kit is designed and produced to be used with Tandem Gold Mass Spectrometer.

-The kit contains enough reagents and materials to analyze 200 samples.

-Zivak LC-MS/MS Organic acid analysis kit can be used in the screening, diagnosis and follow-up of organic acidurias resulting from inborn errors of lipid, carbohydrate, amino acids and steroid metabolism.

-Zivak LC-MS/MS Phenylketonuria analysis kit is produced in accordance with the TS ISO 9001:2000 and TS ISO 13485:2003 quality standards.

INTENDED USE

Organic acidurias are a group of disorders seen due to inborn errors in the metabolisms of lipids, carbohydrates, amino acids, nucleic acids and steroids. These disorders are characterized by the accumulation and increased urinary excretion of some of the metabolites of these molecules. The incidence of these disorders can exhibit a wide variation from 1 in 10,000 to 1,000,000 live births, according to the specific disorder and population discussed. Many of them are inherited autosomal recessively and a prenatal diagnosis method is not available for most of these disorders. If the family of the baby did not have an affected sibling before, there may not be an early warning. The crucial importance of an accurate and prompt diagnosis becomes all the more apparent when the rarity of these disorders, and the facts that these disorders may not show symptoms in the first days of life, and the formation of irreversible damages are taken into consideration.

SAMPLE AND REAGENT PREPARATION PROCEDURE

1. Add 300 mL of urine sample (or calibrators) into sample preparation vials. 
2. Add 700 µL of R1 and vortex for 5 seconds.
3. Inject 10 µL of the above mixtures into the Tandem Gold LC-MS/MS.

The sample will maintain its stability for 24 hours in room temperature, until injected.

MEASUREMENT PARAMETERS

	RT (min)	MH+ (m/z)	MS/MS	Capillary (eV)	CE (eV)
Glikolik acid	2,5	75	75	-30	5
Pirüvik acid	2,7	86,8	86,8	-30	5
Orotik acid	2,8	154,6	110,8	-30	10
Laktik acid	3,1	88,8	88,8	-30	8
Metilmalonik acid	3,8	116,7	73	-30	10
Betahidroksi bütirik acid	3,9	102,8	59	-30	12
Fumarik acid	4,4	114,7	71	-30	8
Süksinik acid	4,4	116,7	73	-30	10
Betahidroksi iso valerik acid	6,1	116,7	59	-30	15
Homogensitik acid	6,6	166,5	122,8	-30	15
Parahidroksi fenil pirüvik acid	7,2	178,5	106,8	-30	10
Etilmalonik acid	7,4	130,7	87	-30	10
Glutarik acid	7,5	130,7	87	-30	10
Adipik acid	8,1	144,5	101	-30	15
Alfahidroksi isovalerik acid	8,5	116,7	71	-30	15
Parahidroksi fenillaktik acid	8,5	180,6	118,8	-30	20
Ortohidroksi fenilasetik acid	9,0	150,6	106,8	-30	15
Parahidroksi fenilasetik acid	9,4	150,6	106,8	-30	15
Suberik acid	9,8	172,6	111	-30	15
Sebaseik acid	11,4	200,7	182,8	-30	15

CHROMATOGRAMS

ORGANİK ASİT LC-MS/MS ANALİZ KİTİ

ZV-3004-0200-20	Organic Acids LC-MS/MS Analysis Kit, Urine  (200 tests)
Reagents
Catalog No:	Product Name	Volume	Quantity
ZV-3004-02S1-20	Calibrator 1 (S1)	3 mL	1 / kit
ZV-3004-02S2-20	Calibrator 2 (S2)	3 mL	1 / kit
ZV-3004-02S3-20	Calibrator 3 (S3)	3 mL	1 / kit
ZV-3004-02R1-20	Reagent 1 (R1)	140 mL	1 / kit
ZV-3004-02MA-20	Mobil Phase A	1 L	1 / kit
ZV-3004-02MB-20	Mobil Phase B	1 L	1 / kit
Materials
ZV-9001-0100	Sample preparation vial(1,8 mL)	100/ pack	2 / kit
ZV-9002-0100	Sample preparation vial cap  (1,8 mL)	100/ pack	2 / kit
ZV-9004-1000	100-1000 µL  pipet tip	100/ pack	1 / kit
ZV-9015-0001	100-1000 µL  pipet		1 / kit
ZV-3004-KK-T Rev.00	User Manual		1 / kit
Materials not supplied
	Vortex mixer
ZV-3004-02C1-20	Zivak- Organic Acid column	2 mL	1 / kit
Storage and Stability
	S1, S2, and S3  should be kept at 4°C R1 should be kept at room temperature. When kept as suggested in the user manual, all reagents will be stable until the expiry date.
Warnings and precautions
	As a general precaution, all sample preparation steps should be carried out in a safety hood wearing protective gloves and glasses. When working with biological samples, necessary steps should be taken to avoid contamination with bloodborne pathogens. All samples posing a biological threat should be appropriately disposed of.

RESULTS

The results are expressed in mg/L. The results can also be expressed as mg organic acid / g creatinine by calculating a ratio of the observed organic acid value to urinary creatinine value in g/L.

PERFORMANCE CHARACTERISTICS

Analytical Sensitivity

	LOD (mg/L)	LOQ (mg/L)
Glycholic acid	0,3	1,0
Pyruvic acid	0,2	0,7
Orotic acid	0,12	0,4
Lactic acid	0,12	0,4
Methylmalonic acid	0,15	0,5
Betahydroxy butyric acid	0,25	0,8
Fumaric acid	0,15	0,5
Succinic acid	0,4	1,3
Betahydroxy iso valeric acid	0,25	0,8
Homogensitic acid	0,13	0,4
Parahydroxy phenyl pyruvic acid	0,3	1,0
Ethylmalonic acid	0,1	0,3
Glutaric acid	0,25	0,8
Adipic acid	0,1	0,3
Alfahydroxy isovaleric acid	0,1	0,3
Parahydroxy phenyllactic acid	0,02	0,1
Orthohydroxy phenylacetic acid	0,3	1,0
Parahydroxy phenylacetic acid	0,15	0,5
Suberic acid	0,05	0,2
Sebacic acid	0,2	0,7

LOD: Limit of Detection

LOQ: Limit of Quantitation

Specificity

The specificty of the method for these organic acids is 100 %

Precision

The organic acid determinations have a CV of 1,9 — 6,7 %.

Linearity

The correlation coefficient (R2) for linearity of organic acid determinations are between 0,995— 0,999.

Interference

The presence of hemoglobin, lipids and bilirubin does not interfere with the method.

Zivak LC-MS/MS Organic acid analysis kit is designed and produced to be used with Tandem Gold Mass Spectromete